JP2024023226A - Dosing regimens for mobilization of hematopoietic stem and progenitor cells - Google Patents

Abstract

To provide compositions and methods useful for mobilizing populations of hematopoietic stem and progenitor cells within a donor, as well as for determining whether samples of mobilized cells are suitable for release for ex vivo expansion and/or therapeutic use.SOLUTION: A method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood is provided, the method comprising administering to the donor (i) a CXCR2 agonist selected from a group consisting of Gro-β, Gro-βT, and variants thereof at a dose of from about 50 μg/kg to about 300 mg/kg and (ii) a CXCR4 antagonist.SELECTED DRAWING: None

Description

[Cross reference to related applications]
This application was filed in U.S. Provisional Application No. 15/834,017, filed on December 6, 2017, and in U.S. Provisional Application No. 62/596,056, filed on December 7, 2017, and Provisional Patent Application No. 16/101,676 filed October 31, 2018;
Claims priority benefit to U.S. Provisional Patent Application No. 62/773,954, filed on November 30,
The disclosures of each of these are incorporated herein by reference in their entirety.

Mobilizing hematopoietic stem and progenitor cells from donors (e.g., human donors) and patients suffering from various pathological conditions (e.g., hematological disorders, metabolic disorders, cancer, and autoimmune diseases, among others) Concerning treating.

Despite advances in medical technology, there remains a need to treat pathologies of the hematopoietic system, such as certain blood cell diseases, metabolic disorders, cancer, and autoimmune conditions, among others. While hematopoietic stem cells have significant therapeutic potential, limitations that prevent their use in the clinic are related to the difficulty in releasing hematopoietic stem cells from the donor's bone marrow into the peripheral blood ( The hematopoietic stem cells are isolated from the donor for infusion into a patient).

A further limitation is that approximately 80% of allogeneic recipients of mobilized peripheral blood (mPB) suffer from graft-versus-host disease.
(graft versus host disease (GVHD)). mPB compared to bone marrow transplantation
Despite higher levels of CD3+ T-cells in the graft, the levels of acute GVHD observed after transplanting HLA-matched mPB are comparable to HLA-matched bone marrow. One explanation is that G-CSF
The grafts mobilized with myeloid-derived suppressor cells, which have immunosuppressive properties that can suppress T-cell proliferation in vitro.
(MDSCs)). The proportion of MDSCs varies in grafts mobilized with G-CSF, and clinical data indicate that patients transplanted with mPB grafts containing more MDSCs have lower rates of acute GVHD, including lower rates of acute GVHD. It has been suggested that it may have better outcomes (Vendramin et al., (2014) BBMT 20(12):2049-2055)
.

Accordingly, there is currently a need for compositions and methods for promoting the mobilization of hematopoietic stem and progenitor cells, particularly for methods of identifying populations of mobilized cells suitable for therapeutic use. There is also a need for compositions and methods for promoting the mobilization of hematopoietic stem and progenitor cells that consistently produce higher numbers of MDSCs than prior art methods.

The present invention provides compositions and methods for mobilizing hematopoietic stem and progenitor cells in a subject. For example, the subject may be a hematopoietic stem and progenitor cell donor (ie, a donor), such as a mammalian donor, particularly a human donor. The invention further provides compositions and methods for treating disorders, such as stem cell disorders, in a patient (eg, a human patient). Gro-β as described herein using the compositions and methods described herein.
or a variant thereof, such as a truncated form of Gro-β (e.g., Gro-β T), optionally a 1,1'-[1,4- Phenylene bis(methylene
)]-bis-1,4,8,11-tetra-azacyclotetradecane or a variant thereof, in combination with a CXC chemokine receptor type 4 (CXCR4) antagonist, in an amount sufficient to mobilize hematopoietic stem and progenitor cells. It may be administered to a subject. Importantly, the compositions and methods described herein can be used to mobilize hematopoietic stem and progenitor cells from a stem cell niche within a donor, such as a human donor, into the circulating peripheral blood of said donor. This means that there is (
On the other hand, it reduces the recruitment of other cells of the hematopoietic lineage, such as white blood cells, neutrophils, lymphocytes, and monocytes). Accordingly, the compositions and methods described herein allow for the selective mobilization of hematopoietic stem and progenitor cells in a donor, which can then be isolated from the donor for therapeutic use. be.

In some embodiments, the hematopoietic stem or progenitor cells may be mobilized from the bone marrow of the donor into peripheral blood, and the hematopoietic stem or progenitor cells may be collected and/or isolated from the peripheral blood. be. Once the mobilized cells are collected, the harvested hematopoietic stem or progenitor cells are then transferred to a patient (who is HLA-compatible with said donor or with said donor for the treatment of one or more conditions of the hematopoietic system). may be injected into another object, such as a sexual object). In some embodiments, the harvested hematopoietic stem or progenitor cells are first grown ex viv before infusing these cells and/or their progeny into the patient.
Grow in o. The compositions and methods described herein enable the production of a population of cells enriched in hematopoietic stem cells compared to other cell types (e.g., leukocytes, neutrophils, and monocytes).
Provides important clinical benefits. Accordingly, populations of mobilized hematopoietic stem and progenitor cells produced using the compositions and methods described herein are particularly suitable for hematopoietic stem cell transplantation therapy and, optionally, for delivery to patients. Ex vivo expansion is performed first to increase the amount of hematopoietic stem and progenitor cells available for injection.

Furthermore, the methods described herein provide the advantage of inhibiting leukocytosis in said donor. Leukocytosis can lead to splenic rupture, renal dysfunction, acute febrile noninfectious pneumonitis (i.e., pulmonary toxicity), cardiovascular toxicity (e.g., hypercoagulability, heart attack, dyspnea, angina, arrhythmias, hypertrophic atherosclerotic plaque rupture due to pro-inflammatory effects associated with bulb count), neurological disorders (e.g.
It can lead to adverse effects such as blurred vision, headaches, and retinal hemorrhages), and sickle cell crisis. For example, D'Souza et al. (2008) Transfusion Medicine Reviews 22(4):28
See 0-290.

As described herein, hematopoietic stem cells are capable of differentiating into multiple cell types in the hematopoietic lineage, and therefore, in order to engraft or repopulate defective or missing cell types in a patient. , may be administered to said patient. The patient is, for example, a patient suffering from one or more blood diseases, such as an autoimmune disease, cancer, hemoglobinopathies, or other hematopoietic pathologies, and thus in need of a hematopoietic stem cell transplant. Sometimes. Accordingly, the present invention particularly relates to sickle cell anemia, thalassemia, Fanconi anemia, Wiskott-Aldrich syndrome, adenosine deaminase deficiency - severe combined immunodeficiency, metachromatic leukodystrophy, Diamond-Blackfan anemia and various hematopoietic conditions such as Wachmann-Diamond syndrome, human immunodeficiency virus infection, and acquired immunodeficiency syndrome;
and cancer and autoimmune diseases.

In a first aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising
agonist and CXCR4 antagonist sufficient to produce a population of cells having a ratio of CD34 ⁺ cells to white blood cells in a sample of peripheral blood of said donor from about 0.0008 to about 0.0021 after administration of said CXCR2 agonist and said CXCR4 antagonist. and administering to said donor in an amount that is suitable for said donor. In some embodiments, the ratio of CD34 ⁺ cells to white blood cells in the sample is about 0.00080, 0.00081, 0.00082, 0.00083, 0.00084, 0.00085, 0.00086, 0.00087,
0.00088, 0.00089, 0.00090, 0.00091, 0.00092, 0.00093, 0.00094, 0.00095, 0.00096,
0.00097, 0.00098, 0.00099, 0.00100, 0.00101, 0.00102, 0.00103, 0.00104, 0.00105
, 0.00106, 0.00107, 0.00108, 0.00109, 0.00110, 0.00111, 0.00112, 0.00113, 0.0011
4, 0.00115, 0.00116, 0.00117, 0.00118, 0.00119, 0.00120, 0.00121, 0.00122, 0.001
23, 0.00124, 0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00
132, 0.00133, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.0
0141, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.
00150, 0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158, 0
.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164, 0.00165, 0.00166, 0.00167,
0.00168, 0.00169, 0.00170, 0.00171, 0.00172, 0.00173, 0.00174, 0.00175, 0.00176,
0.00178, 0.00179, 0.00180, 0.00181, 0.00182, 0.00183, 0.00184, 0.00185, 0.00186
, 0.00187, 0.00188, 0.00189, 0.00190, 0.00191, 0.00192, 0.00193, 0.00194, 0.0019
5, 0.00196, 0.00197, 0.00198, 0.00199, 0.00200, 0.00201, 0.00202, 0.00203, 0.002
04, 0.00205, 0.00206, 0.00207, 0.00208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00
213, 0.00214, 0.00215, 0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.0
0222, 0.00223, 0.00224, or 0.00225. In some embodiments,
The ratio of CD34 ⁺ cells to white blood cells in the sample is about 0.0009 to about 0.002, about 0.001 to about 0.
.0019, about 0.0011 to about 0.0018, about 0.0012 to about 0.0017, about 0.0013 to about 0.0016, or about 0.0014
~0.0015. In some embodiments, the ratio of CD34 ⁺ cells to white blood cells in the sample is about 0.0010 to about 0.0018, e.g., about 0.00100, 0.00101, 0.00102, 0.00103, 0.
00104, 0.00105, 0.00106, 0.00107, 0.00108, 0.00109, 0.00110, 0.00111, 0.00112, 0
.00113, 0.00114, 0.00115, 0.00116, 0.00117, 0.00118, 0.00119, 0.00120, 0.00121,
0.00122, 0.00123, 0.00124, 0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130,
0.00131, 0.00132, 0.00133, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138, 0.00139
, 0.00140, 0.00141, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.0014
8, 0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.001
57, 0.00158, 0.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164, 0.00165, 0.00
166, 0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172, 0.00173, 0.00174, 0.0
C for leukocytes in said sample of 0.0175, 0.00176, 0.00178, 0.00179, or 0.00180.
The proportion of D34 ⁺ cells is, In some embodiments, the CD for leukocytes in said sample
The proportion of ³⁴⁺ cells is about 0.0012 to about 0.0016, e.g. about 0.00120, 0.00121, 0.00122, 0.0012
3, 0.00124, 0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.001
32, 0.00133, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.00
141, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.0
0150, 0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158, 0.
The ratio of CD34 ⁺ cells to white blood cells in said sample is 0.00159, or 0.00160. In some embodiments, the ratio of CD34 ⁺ cells to white blood cells in said sample is about 0.0014.
It is.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising a CXCR2 agonist and a CXCR4 antagonist. , as assessed by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and the CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and the CXCR4 antagonist, about comprising administering to said donor an amount sufficient to enrich said donor's peripheral blood with CD34 ⁺ cells to white blood cells at a ratio of 3.40:1 to about 6.90:1. In some embodiments, the peripheral blood of the donor is about 3.40:1, 3.45:1, 3.50:1, 3.55:1, 3.60:1, 3.65:1, 3.70
:1, 3.75:1, 3.80:1, 3.85:1, 3.90:1, 3.95:1, 4.00:1, 4.05:1, 4.10:1, 4.15:1, 4.20
:1, 4.25:1, 4.30:1, 4.35:1, 4.40:1, 4.45:1, 4.50:1, 4.55:1, 4.60:1, 4.65:1, 4.70
:1, 4.75:1, 4.80:1, 4.85:1, 4.90:1, 4.95:1, 5.00:1, 5.05:1, 5.10:1, 5.15:1, 5.20
:1, 5.25:1, 5.30:1, 5.35:1, 5.40:1, 5.45:1, 5.50:1, 5.55:1, 5.60:1, 5.65:1, 5.70
:1, 5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1, 6.15:1, 6.20
:1, 6.25:1, 6.30:1, 6.35:1, 6.40:1, 6.45:1, 6.50:1, 6.55:1, 6.60:1, 6.65:1, 6.70
It may be enriched with CD34 ⁺ cells to white blood cells to a ratio of :1, 6.75:1, 6.80:1, 6.85:1, or 6.90:11. In some embodiments, the peripheral blood of the donor is about 3.5:1 to about 6.8
:1, approximately 3:1 to approximately 6.7:1, approximately 3.8:1 to approximately 6.6:1, approximately 3:1 to approximately 6.9:1, approximately 4:1 to approximately 6.4:1, approximately 4.1:1 to
Approximately 6.1:1, approximately 4.3:1 to approximately 6.1:1, approximately 4.4:1 to approximately 6:1, approximately 4.5:1 to approximately 6:1, approximately 4.6:1 to approximately 5.9:1, approximately 4
Enrich with CD34 ⁺ cells to leukocytes to a ratio of .7:1 to about 5.8:1, or about 4.8:1 to about 5.7:1. In some embodiments, the peripheral blood of the donor is in a ratio of about 4.0:1 to about 6.0:1, such as about 4.00:1, 4.05:1, 4.10:1, 4.15:1, 4.20:1, 4.25 :1, 4.30:1, 4.35:1, 4.40:1, 4.45
:1, 4.50:1, 4.55:1, 4.60:1, 4.65:1, 4.70:1, 4.75:1, 4.80:1, 4.85:1, 4.90:1, 4.95
:1, 5.00:1, 5.05:1, 5.10:1, 5.15:1, 5.20:1, 5.25:1, 5.30:1, 5.35:1, 5.40:1, 5.45
:1, 5.50:1, 5.55:1, 5.60:1, 5.65:1, 5.70:1, 5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95
Enrich with CD34 ⁺ cells to leukocytes to a ratio of :1, or 6.00:1. In some embodiments, the peripheral blood of the donor is in a ratio of about 4.5:1 to about 5.5:1, e.g., about 4.50:1, 4.55:1.
, 4.60:1, 4.65:1, 4.70:1, 4.75:1, 4.80:1, 4.85:1, 4.90:1, 4.95:1, 5.00:1, 5.05:1
, 5.10:1, 5.15:1, 5.20:1, 5.25:1, 5.30:1, 5.35:1, 5.40:1, 5.45:1, or enriched with CD34 ⁺ cells to leukocytes at a ratio of 5.50 let In some embodiments, the donor's peripheral blood is enriched with CD34 ⁺ cells to leukocytes at a ratio of about 5.1:1.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising at least about
A density of CD34 ⁺ cells of 38,000 cells/ml, e.g., about 38,000 cells/ml to about 100,000 cells/ml, about
40,000 cells/ml to about 90,000 cells/ml, about 50,000 cells/ml to about 80,000 cells/ml, or about 60,
000 cells/ml to approximately 70,000 cells/ml (e.g. approximately 38,00 cells/ml, 39,000 cells/ml, 40,000 cells/ml)
ml, 41,000 cells/ml, 42,000 cells/ml, 43,000 cells/ml, 44,000 cells/ml, 45,000 cells/ml, 4
6,000 cells/ml, 47,000 cells/ml, 48,000 cells/ml, 49,000 cells/ml, 50,000 cells/ml, 51,000
cells/ml, 52,000 cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000 cells/
ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000 cells/ml, 61,000 cells/ml, 6
2,000 cells/ml, 63,000 cells/ml, 64,000 cells/ml, 65,000 cells/ml, 66,000 cells/ml, 67,000
cells/ml, 68,000 cells/ml, 69,000 cells/ml, 70,000 cells/ml, 71,000 cells/ml, 72,000 cells/
ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000 cells/ml, 77,000 cells/ml, 7
8,000 cells/ml, 79,000 cells/ml, 80,000 cells/ml, 81,000 cells/ml, 82,000 cells/ml, 83,000
cells/ml, 84,000 cells/ml, 85,000 cells/ml, 86,000 cells/ml, 87,000 cells/ml, 88,000 cells/
ml, 89,000 cells/ml, 90,000 cells/ml, 91,000 cells/ml, 92,000 cells/ml, 93,000 cells/ml, 9
4,000 cells/ml, 95,000 cells/ml, 96,000 cells/ml, 97,000 cells/ml, 98,000 cells/ml, 99,000
cells/ml, 100,000 cells/ml, or more ⁾ , and approximately 5.3 x 10
A white blood cell density of ⁷ cells/ml or less, such as about 2.3 x 10 ⁷ cells/ml to about 5.3 x 10 ⁷ cells/ml, about 2.
5 x 10 ⁷ cells/ml ~ approx. 5.1 x 10 ⁷ cells/ml, 2.9 x 10 ⁷ cells/ml ~ approx. 4.5 x 10 ⁷ cells/ml, approx. 3 x
10 ⁷ cells/ml to about 4 x 10 ⁷ cells/ml (e.g. 5.3 x 10 ⁷ cells/ml, 5.2 x 10 ⁷ cells/ml, 5.1 x
10 ⁷ cells/ml, 5 x 10 ⁷ cells/ml, 4.9 x 10 ⁷ cells/ml, 4.8 x 10 ⁷ cells/ml, 4.7 x 10 ⁷ cells/ml
, 4.6 x 10 ⁷ cells/ml, 4.5 x 10 ⁷ cells/ml, 4.4 x 10 ⁷ cells/ml, 4.3 x 10 ⁷ cells/ml 4.2 x 1
0 ⁷ cells/ml, 4.1 x 10 ⁷ cells/ml 4 x 10 ⁷ cells/ml, 3.9 x 10 ⁷ cells/ml, 3.8 x 10 ⁷ cells/ml,
3.7 x 10 ⁷ cells/ml, 3.6 x 10 ⁷ cells/ml, 3.5 x 10 ⁷ cells/ml, 3.4 x 10 ⁷ cells/ml, 3.3 x 10
⁷ cells/ml, 3.2 x 10 ⁷ cells/ml, 3.1 x 10 ⁷ cells/ml, 3 x 10 ⁷ cells/ml, 2.9 x 10 ⁷ cells/ml,
2.8 x 10 ⁷ cells/ml, 2.7 x 10 ⁷ cells/ml, 2.6 x 10 ⁷ cells/ml, 2.5 x 10 ⁷ cells/ml, 2.4 x 10
administering to said donor a CXCR2 agonist and a CXCR4 antagonist in an amount sufficient to produce a population of cells having a density of white blood cells of ⁷ cells/ml, 2.3 x 10 ⁷ cells/ml, or less. include. In some embodiments, the method provides between about 38,000 cells/ml and about 100,000 cells/ml.
^{CXCR2 agonist and} administering a CXCR4 antagonist to said donor. In some embodiments, the method provides about 40,000
having a density of CD34 ⁺ cells from about 80,000 cells/ml to about 80,000 cells/ml, and from about 2.5 x 10 ⁷ cells/ml to about 5 x 1
administering to said donor a CXCR2 agonist and a CXCR4 antagonist in amounts sufficient to produce a cell population having a density of ^0.07 cells/ml of white blood cells. In some embodiments, the method has a density of CD34 ⁺ cells of about 50,000 cells/ml to about 90,000 cells/ml, and about 3×
administering to said donor a CXCR2 agonist and a CXCR4 antagonist in an amount sufficient to produce a cell population having a density of white blood cells from 10 ⁷ cells/ml to about 4×10 ⁷ cells/ml.

In a further aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising
agonists and CXCR4 antagonists to produce a population of cells in which the ratio of CD34 ⁺ cells to neutrophils in a sample of peripheral blood of the donor is about 0.0018 to about 0.0058 after administration of the CXCR2 agonist and the CXCR4 antagonist; administering to said donor in an amount sufficient to In some embodiments, the ratio of CD34 ⁺ cells to neutrophils in the sample is about 0.00180, 0.00181, 0.00182, 0.00183, 0.00184, 0.00185, 0.00186, 0.00187, 0.
00188, 0.00189, 0.00190, 0.00191, 0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0
.00197, 0.00198, 0.00199, 0.00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205,
0.00206, 0.00207, 0.00208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00213, 0.00214,
0.00215, 0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.00222, 0.00223
, 0.00224, 0.00225, 0.00226, 0.00227, 0.00228, 0.00229, 0.00230, 0.00231, 0.0023
2, 0.00233, 0.00234, 0.00235, 0.00236, 0.00237, 0.00238, 0.00239, 0.00240, 0.002
41, 0.00242, 0.00243, 0.00244, 0.00245, 0.00246, 0.00247, 0.00248, 0.00249, 0.00
250, 0.00251, 0.00252, 0.00253, 0.00254, 0.00255, 0.00256, 0.00257, 0.00258, 0.0
0259, 0.00260, 0.00261, 0.00262, 0.00263, 0.00264, 0.00265, 0.00266, 0.00267, 0.
00268, 0.00269, 0.00270, 0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0
.00277, 0.00278, 0.00279, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285,
0.00286, 0.00287, 0.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294,
0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.00300, 0.00301, 0.00302
, 0.00303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.0031
1, 0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319, 0.003
20, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00
329, 0.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.00337, 0.0
0338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343, 0.00344, 0.00345, 0.00346, 0.
00347, 0.00348, 0.00349, 0.00350, 0.00351, 0.00352, 0.00353, 0.00354, 0.00355, 0
.00356, 0.00357, 0.00358, 0.00359, 0.00360, 0.00361, 0.00362, 0.00363, 0.00364,
0.00365, 0.00366, 0.00367, 0.00368, 0.00369, 0.00370, 0.00371, 0.00372, 0.00373,
0.00374, 0.00375, 0.00376, 0.00377, 0.00378, 0.00379, 0.00380, 0.00381, 0.00382
, 0.00383, 0.00384, 0.00385, 0.00386, 0.00387, 0.00388, 0.00389, 0.00390, 0.0039
1, 0.00392, 0.00393, 0.00394, 0.00395, 0.00396, 0.00397, 0.00398, 0.00399, 0.004
00, 0.00401, 0.00402, 0.00403, 0.00404, 0.00405, 0.00406, 0.00407, 0.00408, 0.00
409, 0.00410, 0.00411, 0.00412, 0.00413, 0.00414, 0.00415, 0.00416, 0.00417, 0.0
0418, 0.00419, 0.00420, 0.00421, 0.00422, 0.00423, 0.00424, 0.00425, 0.00426, 0.
00427, 0.00428, 0.00429, 0.00430, 0.00431, 0.00432, 0.00433, 0.00434, 0.00435, 0
.00436, 0.00437, 0.00438, 0.00439, 0.00440, 0.00441, 0.00442, 0.00443, 0.00444,
0.00445, 0.00446, 0.00447, 0.00448, 0.00449, 0.00450, 0.00451, 0.00452, 0.00453,
0.00454, 0.00455, 0.00456, 0.00457, 0.00458, 0.00459, 0.00460, 0.00461, 0.00462
, 0.00463, 0.00464, 0.00465, 0.00466, 0.00467, 0.00468, 0.00469, 0.00470, 0.0047
1, 0.00472, 0.00473, 0.00474, 0.00475, 0.00476, 0.00477, 0.00478, 0.00479, 0.004
80, 0.00481, 0.00482, 0.00483, 0.00484, 0.00485, 0.00486, 0.00487, 0.00488, 0.00
489, 0.00490, 0.00491, 0.00492, 0.00493, 0.00494, 0.00495, 0.00496, 0.00497, 0.0
0498, 0.00499, 0.00500, 0.00501, 0.00502, 0.00503, 0.00504, 0.00505, 0.00506, 0.
00507, 0.00508, 0.00509, 0.00510, 0.00511, 0.00512, 0.00513, 0.00514, 0.00515, 0
.00516, 0.00517, 0.00518, 0.00519, 0.00520, 0.00521, 0.00522, 0.00523, 0.00524,
0.00525, 0.00526, 0.00527, 0.00528, 0.00529, 0.00530, 0.00531, 0.00532, 0.00533,
0.00534, 0.00535, 0.00536, 0.00537, 0.00538, 0.00539, 0.00540, 0.00541, 0.00542
, 0.00543, 0.00544, 0.00545, 0.00546, 0.00547, 0.00548, 0.00549, 0.00550, 0.0055
1, 0.00552, 0.00553, 0.00554, 0.00555, 0.00556, 0.00557, 0.00558, 0.00559, 0.005
60, 0.00561, 0.00562, 0.00563, 0.00564, 0.00565, 0.00566, 0.00567, 0.00568, 0.00
569, 0.00570, 0.00571, 0.00572, 0.00573, 0.00574, 0.00575, 0.00576, 0.00577, 0.0
May be 0578, 0.00579, or 0.00580. In some embodiments, the ratio of CD34 ⁺ cells to neutrophils in the sample is about 0.002 to about 0.0056, about 0.0022 to about 0.0054, about
0.0024 to about 0.0052, about 0.0026 to about 0.005, about 0.0028 to about 0.0048, or about 0.003 to about 0.0046
It is. In some embodiments, the ratio of CD34 ⁺ cells to neutrophils in the sample is about 0.0026 to about 0.0046, such as about 0.00260, 0.00261, 0.00262, 0.00263, 0.00264, 0.0
0265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.00272, 0.00273, 0.
00274, 0.00275, 0.00276, 0.00277, 0.00278, 0.00279, 0.00280, 0.00281, 0.00282, 0
.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290, 0.00291,
0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300,
0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308
, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0.0031
7, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325, 0.003
26, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00
335, 0.00336, 0.00337, 0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343, 0.0
0344, 0.00345, 0.00346, 0.00347, 0.00348, 0.00349, 0.00350, 0.00351, 0.00352, 0.
00353, 0.00354, 0.00355, 0.00356, 0.00357, 0.00358, 0.00359, 0.00360, 0.00361, 0
.00362, 0.00363, 0.00364, 0.00365, 0.00366, 0.00367, 0.00368, 0.00369, 0.00370,
0.00371, 0.00372, 0.00373, 0.00374, 0.00375, 0.00376, 0.00377, 0.00378, 0.00379,
0.00380, 0.00381, 0.00382, 0.00383, 0.00384, 0.00385, 0.00386, 0.00387, 0.00388
, 0.00389, 0.00390, 0.00391, 0.00392, 0.00393, 0.00394, 0.00395, 0.00396, 0.0039
7, 0.00398, 0.00399, 0.00400, 0.00401, 0.00402, 0.00403, 0.00404, 0.00405, 0.004
06, 0.00407, 0.00408, 0.00409, 0.00410, 0.00411, 0.00412, 0.00413, 0.00414, 0.00
415, 0.00416, 0.00417, 0.00418, 0.00419, 0.00420, 0.00421, 0.00422, 0.00423, 0.0
0424, 0.00425, 0.00426, 0.00427, 0.00428, 0.00429, 0.00430, 0.00431, 0.00432, 0.
00433, 0.00434, 0.00435, 0.00436, 0.00437, 0.00438, 0.00439, 0.00440, 0.00441, 0
.00442, 0.00443, 0.00444, 0.00445, 0.00446, 0.00447, 0.00448, 0.00449, 0.00450,
0.00451, 0.00452, 0.00453, 0.00454, 0.00455, 0.00456, 0.00457, 0.00458, 0.00459,
or the ratio of CD34 ⁺ cells to neutrophils in said sample of 0.00460. In some embodiments, the ratio of CD34 ⁺ cells to neutrophils in the sample is about 0.0036.

In a further aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising
Agonists and CXCR4 antagonists are evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and the CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and the CXCR4 antagonist. administering to said donor in an amount sufficient to enrich said donor's peripheral blood with CD34 ⁺ cells to neutrophils in a ratio of about 2.1:1 to about 8.1:1 when including. In some embodiments, the peripheral blood of the donor is about 2.10:1, 2.15:1, 2.20:1, 2.25:1, 2.30:1, 2.35:1, 2.40
:1, 2.45:1, 2.50:1, 2.55:1, 2.60:1, 2.65:1, 2.70:1, 2.75:1, 2.80:1, 2.85:1, 2.90
:1, 2.95:1, 3.00:1, 3.05:1, 3.10:1, 3.15:1, 3.20:1, 3.25:1, 3.30:1, 3.35:1, 3.40
:1, 3.45:1, 3.50:1, 3.55:1, 3.60:1, 3.65:1, 3.70:1, 3.75:1, 3.80:1, 3.85:1, 3.90
:1, 3.95:1, 4.00:1, 4.05:1, 4.10:1, 4.15:1, 4.20:1, 4.25:1, 4.30:1, 4.35:1, 4.40
:1, 4.45:1, 4.50:1, 4.55:1, 4.60:1, 4.65:1, 4.70:1, 4.75:1, 4.80:1, 4.85:1, 4.90
:1, 4.95:1, 5.00:1, 5.05:1, 5.10:1, 5.15:1, 5.20:1, 5.25:1, 5.30:1, 5.35:1, 5.40
:1, 5.45:1, 5.50:1, 5.55:1, 5.60:1, 5.65:1, 5.70:1, 5.75:1, 5.80:1, 5.85:1, 5.90
:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1, 6.15:1, 6.20:1, 6.25:1, 6.30:1, 6.35:1, 6.40
:1, 6.45:1, 6.50:1, 6.55:1, 6.60:1, 6.65:1, 6.70:1, 6.75:1, 6.80:1, 6.85:1, 6.90
:1, 6.95:1, 7.00:1, 7.05:1, 7.10:1, 7.15:1, 7.20:1, 7.25:1, 7.30:1, 7.35:1, 7.40
:1, 7.45:1, 7.50:1, 7.55:1, 7.60:1, 7.65:1, 7.70:1, 7.75:1, 7.80:1, 7.85:1, 7.90
:1, 7.95:1, or 8.00:1 ratios may be enriched with CD34 ⁺ cells to neutrophils. In some embodiments, the peripheral blood of the donor is about 2.5:1 to about 7:1, about 2.6:1 to about 6.9:1
, about 2.7:1 to about 6.8:1, about 2.8:1 to about 6.7:1, about 2.9:1 to about 6.6:1, about 3:1 to about 6.5:1, about 3.2:1
~about 6.4:1, about 3.3:1 to about 6.3:1, about 3.4:1 to about 6.2:1, or about 3.5:1 to about 6.1:1 ratio of CD34 ⁺ cells to neutrophils , enrich. In some embodiments, the peripheral blood of the donor is in a ratio of about 5.4:1 to about 7.4:1, e.g., about 5.40:1, 5.45:1, 5.50:1, 5.55:1, 5.60:1,
5.65:1, 5.70:1, 5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1,
6.15:1, 6.20:1, 6.25:1, 6.30:1, 6.35:1, 6.40:1, 6.45:1, 6.50:1, 6.55:1, 6.60:1,
6.65:1, 6.70:1, 6.75:1, 6.80:1, 6.85:1, 6.90:1, 6.95:1, 7.00:1, 7.05:1, 7.10:1,
CD34 ⁺ to neutrophils in a ratio of 7.15:1, 7.20:1, 7.25:1, 7.30:1, 7.35:1, or 7.40:1
Enrich with cells. In some embodiments, the donor's peripheral blood is enriched with CD34 ⁺ cells to neutrophils at a ratio of about 6.4:1.

In yet another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising:
2 agonist and a CXCR4 antagonist to produce a population of cells in which the ratio of CD34 ⁺ cells to lymphocytes in the peripheral blood sample of the donor is about 0.0021 to about 0.0094 after administration of the CXCR2 agonist and the CXCR4 antagonist. and administering to said donor in an amount sufficient to In some embodiments, the ratio of CD34 ⁺ cells to lymphocytes in the sample is about 0.00210, 0.00211, 0.00212, 0.00213, 0.00214, 0.00215, 0.00216, 0.002
17, 0.00218, 0.00219, 0.00220, 0.00221, 0.00222, 0.00223, 0.00224, 0.00225, 0.00
226, 0.00227, 0.00228, 0.00229, 0.00230, 0.00231, 0.00232, 0.00233, 0.00234, 0.0
0235, 0.00236, 0.00237, 0.00238, 0.00239, 0.00240, 0.00241, 0.00242, 0.00243, 0.
00244, 0.00245, 0.00246, 0.00247, 0.00248, 0.00249, 0.00250, 0.00251, 0.00252, 0
.00253, 0.00254, 0.00255, 0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.00261,
0.00262, 0.00263, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270,
0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00277, 0.00278, 0.00279
, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.0028
8, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.002
97, 0.00298, 0.00299, 0.00300, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00
305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313, 0.0
0314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0.
00323, 0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331, 0
.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339, 0.00340,
0.00341, 0.00342, 0.00343, 0.00344, 0.00345, 0.00346, 0.00347, 0.00348, 0.00349,
0.00350, 0.00351, 0.00352, 0.00353, 0.00354, 0.00355, 0.00356, 0.00357, 0.00358
, 0.00359, 0.00360, 0.00361, 0.00362, 0.00363, 0.00364, 0.00365, 0.00366, 0.0036
7, 0.00368, 0.00369, 0.00370, 0.00371, 0.00372, 0.00373, 0.00374, 0.00375, 0.003
76, 0.00377, 0.00378, 0.00379, 0.00380, 0.00381, 0.00382, 0.00383, 0.00384, 0.00
385, 0.00386, 0.00387, 0.00388, 0.00389, 0.00390, 0.00391, 0.00392, 0.00393, 0.0
0394, 0.00395, 0.00396, 0.00397, 0.00398, 0.00399, 0.00400, 0.00401, 0.00402, 0.
00403, 0.00404, 0.00405, 0.00406, 0.00407, 0.00408, 0.00409, 0.00410, 0.00411, 0
.00412, 0.00413, 0.00414, 0.00415, 0.00416, 0.00417, 0.00418, 0.00419, 0.00420,
0.00421, 0.00422, 0.00423, 0.00424, 0.00425, 0.00426, 0.00427, 0.00428, 0.00429,
0.00430, 0.00431, 0.00432, 0.00433, 0.00434, 0.00435, 0.00436, 0.00437, 0.00438
, 0.00439, 0.00440, 0.00441, 0.00442, 0.00443, 0.00444, 0.00445, 0.00446, 0.0044
7, 0.00448, 0.00449, 0.00450, 0.00451, 0.00452, 0.00453, 0.00454, 0.00455, 0.004
56, 0.00457, 0.00458, 0.00459, 0.00460, 0.00461, 0.00462, 0.00463, 0.00464, 0.00
465, 0.00466, 0.00467, 0.00468, 0.00469, 0.00470, 0.00471, 0.00472, 0.00473, 0.0
0474, 0.00475, 0.00476, 0.00477, 0.00478, 0.00479, 0.00480, 0.00481, 0.00482, 0.
00483, 0.00484, 0.00485, 0.00486, 0.00487, 0.00488, 0.00489, 0.00490, 0.00491, 0
.00492, 0.00493, 0.00494, 0.00495, 0.00496, 0.00497, 0.00498, 0.00499, 0.00500,
0.00501, 0.00502, 0.00503, 0.00504, 0.00505, 0.00506, 0.00507, 0.00508, 0.00509,
0.00510, 0.00511, 0.00512, 0.00513, 0.00514, 0.00515, 0.00516, 0.00517, 0.00518
, 0.00519, 0.00520, 0.00521, 0.00522, 0.00523, 0.00524, 0.00525, 0.00526, 0.0052
7, 0.00528, 0.00529, 0.00530, 0.00531, 0.00532, 0.00533, 0.00534, 0.00535, 0.005
36, 0.00537, 0.00538, 0.00539, 0.00540, 0.00541, 0.00542, 0.00543, 0.00544, 0.00
545, 0.00546, 0.00547, 0.00548, 0.00549, 0.00550, 0.00551, 0.00552, 0.00553, 0.0
0554, 0.00555, 0.00556, 0.00557, 0.00558, 0.00559, 0.00560, 0.00561, 0.00562, 0.
00563, 0.00564, 0.00565, 0.00566, 0.00567, 0.00568, 0.00569, 0.00570, 0.00571, 0
.00572, 0.00573, 0.00574, 0.00575, 0.00576, 0.00577, 0.00578, 0.00579, 0.00580,
0.00581, 0.00582, 0.00583, 0.00584, 0.00585, 0.00586, 0.00587, 0.00588, 0.00589,
0.00590, 0.00591, 0.00592, 0.00593, 0.00594, 0.00595, 0.00596, 0.00597, 0.00598
, 0.00599, 0.00600, 0.00601, 0.00602, 0.00603, 0.00604, 0.00605, 0.00606, 0.0060
7, 0.00608, 0.00609, 0.00610, 0.00611, 0.00612, 0.00613, 0.00614, 0.00615, 0.006
16, 0.00617, 0.00618, 0.00619, 0.00620, 0.00621, 0.00622, 0.00623, 0.00624, 0.00
625, 0.00626, 0.00627, 0.00628, 0.00629, 0.00630, 0.00631, 0.00632, 0.00633, 0.0
0634, 0.00635, 0.00636, 0.00637, 0.00638, 0.00639, 0.00640, 0.00641, 0.00642, 0.
00643, 0.00644, 0.00645, 0.00646, 0.00647, 0.00648, 0.00649, 0.00650, 0.00651, 0
.00652, 0.00653, 0.00654, 0.00655, 0.00656, 0.00657, 0.00658, 0.00659, 0.00660,
0.00661, 0.00662, 0.00663, 0.00664, 0.00665, 0.00666, 0.00667, 0.00668, 0.00669,
0.00670, 0.00671, 0.00672, 0.00673, 0.00674, 0.00675, 0.00676, 0.00677, 0.00678
, 0.00679, 0.00680, 0.00681, 0.00682, 0.00683, 0.00684, 0.00685, 0.00686, 0.0068
7, 0.00688, 0.00689, 0.00690, 0.00691, 0.00692, 0.00693, 0.00694, 0.00695, 0.006
96, 0.00697, 0.00698, 0.00699, 0.00700, 0.00701, 0.00702, 0.00703, 0.00704, 0.00
705, 0.00706, 0.00707, 0.00708, 0.00709, 0.00710, 0.00711, 0.00712, 0.00713, 0.0
0714, 0.00715, 0.00716, 0.00717, 0.00718, 0.00719, 0.00720, 0.00721, 0.00722, 0.
00723, 0.00724, 0.00725, 0.00726, 0.00727, 0.00728, 0.00729, 0.00730, 0.00731, 0
.00732, 0.00733, 0.00734, 0.00735, 0.00736, 0.00737, 0.00738, 0.00739, 0.00740,
0.00741, 0.00742, 0.00743, 0.00744, 0.00745, 0.00746, 0.00747, 0.00748, 0.00749,
0.00750, 0.00751, 0.00752, 0.00753, 0.00754, 0.00755, 0.00756, 0.00757, 0.00758
, 0.00759, 0.00760, 0.00761, 0.00762, 0.00763, 0.00764, 0.00765, 0.00766, 0.0076
7, 0.00768, 0.00769, 0.00770, 0.00771, 0.00772, 0.00773, 0.00774, 0.00775, 0.007
76, 0.00777, 0.00778, 0.00779, 0.00780, 0.00781, 0.00782, 0.00783, 0.00784, 0.00
785, 0.00786, 0.00787, 0.00788, 0.00789, 0.00790, 0.00791, 0.00792, 0.00793, 0.0
0794, 0.00795, 0.00796, 0.00797, 0.00798, 0.00799, 0.00800, 0.00801, 0.00802, 0.
00803, 0.00804, 0.00805, 0.00806, 0.00807, 0.00808, 0.00809, 0.00810, 0.00811, 0
.00812, 0.00813, 0.00814, 0.00815, 0.00816, 0.00817, 0.00818, 0.00819, 0.00820,
0.00821, 0.00822, 0.00823, 0.00824, 0.00825, 0.00826, 0.00827, 0.00828, 0.00829,
0.00830, 0.00831, 0.00832, 0.00833, 0.00834, 0.00835, 0.00836, 0.00837, 0.00838
, 0.00839, 0.00840, 0.00841, 0.00842, 0.00843, 0.00844, 0.00845, 0.00846, 0.0084
7, 0.00848, 0.00849, 0.00850, 0.00851, 0.00852, 0.00853, 0.00854, 0.00855, 0.008
56, 0.00857, 0.00858, 0.00859, 0.00860, 0.00861, 0.00862, 0.00863, 0.00864, 0.00
865, 0.00866, 0.00867, 0.00868, 0.00869, 0.00870, 0.00871, 0.00872, 0.00873, 0.0
0874, 0.00875, 0.00876, 0.00877, 0.00878, 0.00879, 0.00880, 0.00881, 0.00882, 0.
00883, 0.00884, 0.00885, 0.00886, 0.00887, 0.00888, 0.00889, 0.00890, 0.00891, 0
.00892, 0.00893, 0.00894, 0.00895, 0.00896, 0.00897, 0.00898, 0.00899, 0.00900,
0.00901, 0.00902, 0.00903, 0.00904, 0.00905, 0.00906, 0.00907, 0.00908, 0.00909,
0.00910, 0.00911, 0.00912, 0.00913, 0.00914, 0.00915, 0.00916, 0.00917, 0.00918
, 0.00919, 0.00920, 0.00921, 0.00922, 0.00923, 0.00924, 0.00925, 0.00926, 0.0092
7, 0.00928, 0.00929, 0.00930, 0.00931, 0.00932, 0.00933, 0.00934, 0.00935, 0.009
36, 0.00937, 0.00938, 0.00939, or 0.00940. In some embodiments, the ratio of CD34 ⁺ cells to lymphocytes in the sample is about 0.0022 to about 0.0093,
Approx. 0.0023 to approx. 0.0092, approx. 0.0024 to approx. 0.0091, approx. 0.003 to approx. 0.0085, approx. 0.0035 to approx. 0.0075,
or about 0.0045 to about 0.0065. In some embodiments, the ratio of CD34 ⁺ cells to lymphocytes in the sample is about 0.0025 to about 0.0035, such as about 0.00250, 0.00251, 0.00252.
, 0.00253, 0.00254, 0.00255, 0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.0026
1, 0.00262, 0.00263, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.002
70, 0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00277, 0.00278, 0.00
279, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.0
0288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.
00297, 0.00298, 0.00299, 0.00300, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0
.00305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313,
0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322,
0.00323, 0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331
, 0.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339, 0.0034
0, 0.00341, 0.00342, 0.00343, 0.00344, 0.00345, 0.00346, 0.00347, 0.00348, 0.003
49, or the ratio of CD34 ⁺ cells to lymphocytes in the sample of 0.00350. In some embodiments, the ratio of CD34 ⁺ cells to lymphocytes in the sample is about 0.0031.

In a further aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising
Agonists and CXCR4 antagonists are evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and the CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and the CXCR4 antagonist. administering to said donor in an amount sufficient to enrich said donor's peripheral blood with CD34 ⁺ cells to lymphocytes in a ratio of about 4.8:1 to about 8.4:1 when include. In some embodiments, the peripheral blood of the donor is about 4.80:1, 4.85:1, 4.90:1, 4.95:1, 5.00:1, 5.05:1, 5.
10:1, 5.15:1, 5.20:1, 5.25:1, 5.30:1, 5.35:1, 5.40:1, 5.45:1, 5.50:1, 5.55:1, 5.
60:1, 5.65:1, 5.70:1, 5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.
10:1, 6.15:1, 6.20:1, 6.25:1, 6.30:1, 6.35:1, 6.40:1, 6.45:1, 6.50:1, 6.55:1, 6.
60:1, 6.65:1, 6.70:1, 6.75:1, 6.80:1, 6.85:1, 6.90:1, 6.95:1, 7.00:1, 7.05:1, 7.
10:1, 7.15:1, 7.20:1, 7.25:1, 7.30:1, 7.35:1, 7.40:1, 7.45:1, 7.50:1, 7.55:1, 7.
60:1, 7.65:1, 7.70:1, 7.75:1, 7.80:1, 7.85:1, 7.90:1, 7.95:1, 8.00:1, 8.05:1, 8.
It may be enriched with CD34 ⁺ cells to lymphocytes to a ratio of 10:1, 8.15:1, 8.20:1, 8.25:1, 8.30:1, 8.35:1, or 8.40:1. In some embodiments, the peripheral blood of the donor is infused with lymphocytes in a ratio of about 5:1 to about 7:1, about 5.5:1 to about 6.5:1, or about 5.2:1 to about 5.7:1. Enrich against CD34 ⁺ cells. In some embodiments, the peripheral blood of said donor is about 5
.0:1 to about 6.5:1 ratios, such as about 5.00:1, 5.05:1, 5.10:1, 5.15:1, 5.20:1, 5.25:1,
5.30:1, 5.35:1, 5.40:1, 5.45:1, 5.50:1, 5.55:1, 5.60:1, 5.65:1, 5.70:1, 5.75:1,
5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1, 6.15:1, 6.20:1, 6.25:1,
Enrich with CD34 ⁺ cells to lymphocytes to a ratio of 6.30:1, 6.35:1, 6.40:1, 6.45:1, or 6.50:1. In some embodiments, the peripheral blood of the donor is about
Enrich with CD34 ⁺ cells to a 5.7:1 ratio.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising at least about
A density of CD34 ⁺ cells of 38,000 cells/ml, e.g., about 38,000 cells/ml to about 100,000 cells/ml, about
40,000 cells/ml to about 90,000 cells/ml, about 50,000 cells/ml to about 80,000 cells/ml, or about 60,
000 cells/ml to approximately 70,000 cells/ml (e.g. approximately 38,00 cells/ml, 39,000 cells/ml, 40,000 cells/ml)
ml, 41,000 cells/ml, 42,000 cells/ml, 43,000 cells/ml, 44,000 cells/ml, 45,000 cells/ml, 4
6,000 cells/ml, 47,000 cells/ml, 48,000 cells/ml, 49,000 cells/ml, 50,000 cells/ml, 51,000
cells/ml, 52,000 cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000 cells/
ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000 cells/ml, 61,000 cells/ml, 6
2,000 cells/ml, 63,000 cells/ml, 64,000 cells/ml, 65,000 cells/ml, 66,000 cells/ml, 67,000
cells/ml, 68,000 cells/ml, 69,000 cells/ml, 70,000 cells/ml, 71,000 cells/ml, 72,000 cells/
ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000 cells/ml, 77,000 cells/ml, 7
8,000 cells/ml, 79,000 cells/ml, 80,000 cells/ml, 81,000 cells/ml, 82,000 cells/ml, 83,000
cells/ml, 84,000 cells/ml, 85,000 cells/ml, 86,000 cells/ml, 87,000 cells/ml, 88,000 cells/
ml, 89,000 cells/ml, 90,000 cells/ml, 91,000 cells/ml, 92,000 cells/ml, 93,000 cells/ml, 9
4,000 cells/ml, 95,000 cells/ml, 96,000 cells/ml, 97,000 cells/ml, 98,000 cells/ml, 99,000
cells/ml, 100,000 cells/ml, or more ⁾ , and approximately 2.4 x 10
A density of lymphocytes below ⁷ cells/ml, such as from about 1 x 10 ⁷ cells/ml to about 2.3 x 10 ⁷ cells/ml, about 1.
3 x 10 ⁷ cells/ml to about 2.1 x 10 ⁷ cells/ml, or about 1.5 x 10 ⁷ cells/ml to about 1.9 x 10 ⁷ cells/ml
(For example, about 2.4 x 10 ⁷ cells/ml, 2.3 x 10 ⁷ cells/ml, 2.2 x 0 ⁷ cells/ml, 2.1 x 10 ⁷ cells/m
l, 2 x 10 ⁷ cells/ml, 1.9 x 10 ⁷ cells/ml, 1.8 x 10 ⁷ cells/ml, 1.7 x 10 ⁷ cells/ml, 1.6 x 1
0 ⁷ cells/ml, 1.5 x 10 ⁷ cells/ml 1.4 x 10 ⁷ cells/ml, 1.3 x 10 ⁷ cells/ml, 1.2 x 10 ⁷ cells/ml
, 1.1 x 10 ⁷ cells/ml, 1 x 10 ⁷ cells/ml, or less, 0.9 x 10 ⁷ cells/ml, 0.8 x
administering to said donor a CXCR2 agonist and a CXCR4 antagonist in amounts sufficient to produce a population of cells having a density of lymphocytes of 10 ⁷ cells/ml, or less. In some embodiments, the method has a density of CD34 ⁺ cells of about 38,000 cells/ml to about 100,000 cells/ml, and of about 1×10 ⁷ cells/ml to about 2.3×10 ⁷ cells/ml. administering to said donor a CXCR2 agonist and a CXCR4 antagonist in an amount sufficient to produce a population of cells having a density of lymphocytes. In some embodiments, the method provides between about 40,000 cells/ml and about 80,
000 cells/ml and a density of lymphocytes from about 1.3×10 ⁷ cells/ ^ml to about 2.3×10 ⁷ cells/ml, CXCR2 agonists and CXCR4
administering an antagonist to said donor. In some embodiments, the method has a density of CD34 ⁺ cells of about 50,000 cells/ml to about 90,000 cells/ml, and about 1.5×10 ⁷ cells/ml.
CXC in an amount sufficient to produce a cell population with a density of lymphocytes from ml to approximately 2 x 10 ⁷ cells/ml.
administering to said donor an R2 agonist and a CXCR4 antagonist.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising a CXCR2 agonist and a CXCR4 antagonist. , after administration of said CXCR2 agonist and said CXCR4 antagonist, the ratio of CD34 ⁺ cells to monocytes in a sample of peripheral blood of said donor is about 0.0.
0.071 to about 0.0174 to about 0.0174. In some embodiments, the ratio of CD34 ⁺ cells to monocytes in the sample is:
Approximately 0.00710, 0.00711, 0.00712, 0.00713, 0.00714, 0.00715, 0.00716, 0.00717, 0.0071
8, 0.00719, 0.00720, 0.00721, 0.00722, 0.00723, 0.00724, 0.00725, 0.00726, 0.007
27, 0.00728, 0.00729, 0.00730, 0.00731, 0.00732, 0.00733, 0.00734, 0.00735, 0.00
736, 0.00737, 0.00738, 0.00739, 0.00740, 0.00741, 0.00742, 0.00743, 0.00744, 0.0
0745, 0.00746, 0.00747, 0.00748, 0.00749, 0.00750, 0.00751, 0.00752, 0.00753, 0.
00754, 0.00755, 0.00756, 0.00757, 0.00758, 0.00759, 0.00760, 0.00761, 0.00762, 0
.00763, 0.00764, 0.00765, 0.00766, 0.00767, 0.00768, 0.00769, 0.00770, 0.00771,
0.00772, 0.00773, 0.00774, 0.00775, 0.00776, 0.00777, 0.00778, 0.00779, 0.00780,
0.00781, 0.00782, 0.00783, 0.00784, 0.00785, 0.00786, 0.00787, 0.00788, 0.00789
, 0.00790, 0.00791, 0.00792, 0.00793, 0.00794, 0.00795, 0.00796, 0.00797, 0.0079
8, 0.00799, 0.00800, 0.00801, 0.00802, 0.00803, 0.00804, 0.00805, 0.00806, 0.008
07, 0.00808, 0.00809, 0.00810, 0.00811, 0.00812, 0.00813, 0.00814, 0.00815, 0.00
816, 0.00817, 0.00818, 0.00819, 0.00820, 0.00821, 0.00822, 0.00823, 0.00824, 0.0
0825, 0.00826, 0.00827, 0.00828, 0.00829, 0.00830, 0.00831, 0.00832, 0.00833, 0.
00834, 0.00835, 0.00836, 0.00837, 0.00838, 0.00839, 0.00840, 0.00841, 0.00842, 0
.00843, 0.00844, 0.00845, 0.00846, 0.00847, 0.00848, 0.00849, 0.00850, 0.00851,
0.00852, 0.00853, 0.00854, 0.00855, 0.00856, 0.00857, 0.00858, 0.00859, 0.00860,
0.00861, 0.00862, 0.00863, 0.00864, 0.00865, 0.00866, 0.00867, 0.00868, 0.00869
, 0.00870, 0.00871, 0.00872, 0.00873, 0.00874, 0.00875, 0.00876, 0.00877, 0.0087
8, 0.00879, 0.00880, 0.00881, 0.00882, 0.00883, 0.00884, 0.00885, 0.00886, 0.008
87, 0.00888, 0.00889, 0.00890, 0.00891, 0.00892, 0.00893, 0.00894, 0.00895, 0.00
896, 0.00897, 0.00898, 0.00899, 0.00900, 0.00901, 0.00902, 0.00903, 0.00904, 0.0
0905, 0.00906, 0.00907, 0.00908, 0.00909, 0.00910, 0.00911, 0.00912, 0.00913, 0.
00914, 0.00915, 0.00916, 0.00917, 0.00918, 0.00919, 0.00920, 0.00921, 0.00922, 0
.00923, 0.00924, 0.00925, 0.00926, 0.00927, 0.00928, 0.00929, 0.00930, 0.00931,
0.00932, 0.00933, 0.00934, 0.00935, 0.00936, 0.00937, 0.00938, 0.00939, 0.00940,
0.00941, 0.00942, 0.00943, 0.00944, 0.00945, 0.00946, 0.00947, 0.00948, 0.00949
, 0.00950, 0.00951, 0.00952, 0.00953, 0.00954, 0.00955, 0.00956, 0.00957, 0.0095
8, 0.00959, 0.00960, 0.00961, 0.00962, 0.00963, 0.00964, 0.00965, 0.00966, 0.009
67, 0.00968, 0.00969, 0.00970, 0.00971, 0.00972, 0.00973, 0.00974, 0.00975, 0.00
976, 0.00977, 0.00978, 0.00979, 0.00980, 0.00981, 0.00982, 0.00983, 0.00984, 0.0
0985, 0.00986, 0.00987, 0.00988, 0.00989, 0.00990, 0.00991, 0.00992, 0.00993, 0.
00994, 0.00995, 0.00996, 0.00997, 0.00998, 0.00999, 0.0100, 0.0101, 0.0103, 0.01
04, 0.0105, 0.0106, 0.0107, 0.0108, 0.0109, 0.0110, 0.0111, 0.0112, 0.0113, 0.01
14, 0.0115, 0.0116, 0.0117, 0.0118, 0.0119, 0.0120, 0.0121, 0.0122, 0.0123, 0.01
24, 0.0125, 0.0126, 0.0127, 0.0128, 0.0129, 0.0130, 0.0131, 0.0132, 0.0133, 0.01
34, 0.0135, 0.0136, 0.0137, 0.0138, 0.0139, 0.0140, 0.0141, 0.0142, 0.0143, 0.01
44, 0.0145, 0.0146, 0.0147, 0.0148, 0.0149, 0.0150, 0.0151, 0.0152, 0.0153, 0.01
54, 0.0155, 0.0156, 0.0157, 0.0158, 0.0159, 0.0160, 0.0161, 0.0162, 0.0163, 0.01
64, 0.0165, 0.0166, 0.0167, 0.0168, 0.0169, 0.0170, 0.0171, 0.0172, 0.0173, or
0.0174, sometimes. In some embodiments, C for monocytes in said sample
The proportion of D34 ⁺ cells is 0.008 to about 0.016, about 0.009 to about 0.015, about 0.01 to about 0.014, or about 0.011
~about 0.013. In some embodiments, the ratio of CD34 ⁺ cells to monocytes in the sample is about 0.0100 to about 0.0140, such as about 0.0100, 0.0101, 0.0103, 0.0104, 0.0105, 0.
0106, 0.0107, 0.0108, 0.0109, 0.0110, 0.0111, 0.0112, 0.0113, 0.0114, 0.0115, 0.
0116, 0.0117, 0.0118, 0.0119, 0.0120, 0.0121, 0.0122, 0.0123, 0.0124, 0.0125, 0.
0126, 0.0127, 0.0128, 0.0129, 0.0130, 0.0131, 0.0132, 0.0133, 0.0134, 0.0135, 0.
The ratio of CD34 ⁺ cells to monocytes in the sample is 0.0136, 0.0137, 0.0138, 0.0139, or 0.0140. In some embodiments, the ratio of CD34 ⁺ cells to monocytes in said sample is about 0.0118.

In a further aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising: CD34 ⁺ cell density, e.g., from about 38,000 cells/ml to about 100,000 cells/ml,
About 40,000 cells/ml to about 90,000 cells/ml, about 50,000 cells/ml to about 80,000 cells/ml, or about
60,000 cells/ml to approximately 70,000 cells/ml (e.g. approximately 38,00 cells/ml, 39,000 cells/ml, 40,000 cells/ml, 41,000 cells/ml, 42,000 cells/ml, 43,000 cells/ml, 44,000 cells/ml , 45,000 cells/ml
, 46,000 cells/ml, 47,000 cells/ml, 48,000 cells/ml, 49,000 cells/ml, 50,000 cells/ml, 51,
000 cells/ml, 52,000 cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000 cells/ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000 cells/ml, 61,000 cells/ml
, 62,000 cells/ml, 63,000 cells/ml, 64,000 cells/ml, 65,000 cells/ml, 66,000 cells/ml, 67,
000 cells/ml, 68,000 cells/ml, 69,000 cells/ml, 70,000 cells/ml, 71,000 cells/ml, 72,000 cells/ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000 cells/ml, 77,000 cells/ml
, 78,000 cells/ml, 79,000 cells/ml, 80,000 cells/ml, 81,000 cells/ml, 82,000 cells/ml, 83,
000 cells/ml, 84,000 cells/ml, 85,000 cells/ml, 86,000 cells/ml, 87,000 cells/ml, 88,000 cells/ml, 89,000 cells/ml, 90,000 cells/ml, 91,000 cells/ml, 92,000 cells/ml, 93,000 cells/ml
, 94,000 cells/ml, 95,000 cells/ml, 96,000 cells/ml, 97,000 cells/ml, 98,000 cells/ml, 99,
000 cells/ml, 100,000 cells/ml, or more), ^and approximately 6 x 1
Density of monocytes below 0 ⁶ cells/ml, e.g. 3.4 x 10 ⁶ cells/ml to approx. 5.9 x 10 ⁶ cells/ml, approx. 3.5 x
10 ⁶ cells/ml to about 5.7 x 10 ⁶ cells/ml, or about 4 x 10 ⁶ cells/ml to about 5 x 10 ⁶ cells/ml (e.g., 5.9 x 10 ⁶ cells/ml, 5.8 x 10 ⁶ cells/ml) ml, 5.7 x 10 ⁶ cells/ml, 5.6 x 10 ⁶ cells/ml, 5.5 x
10 ⁶ cells/ml, 5.4 x 10 ⁶ cells/ml, 5.3 x 10 ⁶ cells/ml, 5.2 x 10 ⁶ cells/ml, 5.1 x 10 ⁶ cells/ml
ml, 5 x 10 ⁶ cells/ml, 4.9 x 10 ⁶ cells/ml, 4.8 x 10 ⁶ cells/ml, 4.7 x 10 ⁶ cells/ml, 4.6 x
10 ⁶ cells/ml, 4.5 x 10 ⁶ cells/ml, 4.4 x 10 ⁶ cells/ml, 4.3 x 10 ⁶ cells/ml, 4.2 x 10 ⁶ cells/ml
ml, 4.1 x 10 ⁶ cells/ml, 4 x 10 ⁶ cells/ml, 3.9 x 10 ⁶ cells/ml, 3.8 x 10 ⁶ cells/ml, 3.7 x
10 ⁶ cells/ml, 3.6 x 10 ⁶ cells/ml, 3.5 x 10 ⁶ cells/ml, 3.4 x 10 ⁶ cells/ml, or less
CXCR2 agonist and CX in an amount sufficient to produce a population of cells with a density of monocytes of
administering a CR4 antagonist to said donor. In some embodiments, the method has a density of CD34 ⁺ cells of about 38,000 cells/ml to about 100,000 cells/ml, and about 3.4×10 ⁶
CX in an amount sufficient to produce a cell population with a density of monocytes from about 6 x 10 cells/ml to about ⁶ x 10 cells/ml.
administering to said donor a CR2 agonist and a CXCR4 antagonist. In some embodiments, the method has a density of CD34 ⁺ cells of about 40,000 cells/ml to about 80,000 cells/ml, and of about 4×10 ⁶ cells/ml to about 5.5×10 ⁶ cells/ml. administering to said donor a CXCR2 agonist and a CXCR4 antagonist in amounts sufficient to produce a population of cells having a density of monocytes. In some embodiments, the method provides between about 50,000 cells/ml and about 90,000 cells/ml of CD34
CXCR2 agonists and CXCR4 antagonists in amounts sufficient to produce a population of cells having a density of ⁺ cells and a density of monocytes of about 4 x 10 ⁶ cells/ml to about 5 x 10 ⁶ cells/ml. and administering to said donor.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising a CXCR2 agonist and a CXCR4 antagonist. , as assessed by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and the CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and the CXCR4 antagonist, about comprising administering to said donor an amount sufficient to enrich said donor's peripheral blood with CD34 ⁺ cells to monocytes at a ratio of 1.1:1 to about 2.3:1. In some embodiments,
The peripheral blood of the donor is approximately 1.10:1, 1.15:1, 1.20:1, 1.25:1, 1.30:1, 1.35:1, 1.40:1,
1.45:1, 1.50:1, 1.55:1, 1.60:1, 1.65:1, 1.70:1, 1.75:1, 1.80:1, 1.85:1, 1.90:1,
Enrich with CD34 ⁺ cells to monocytes to a ratio of 1.95:1, 2.00:1, 2.05:1, 2.10:1, 2.15:1, 2.20:1, 2.25:1, or 2.30:1. There is. In some embodiments, the peripheral blood of the donor is in a ratio of about 1.3:1 to about 1.9:1, e.g., about 1.30:1, 1.35:1, 1.40:1, 1.45:1,
CD34 ⁺ cells to monocytes at a ratio of 1.50:1, 1.55:1, 1.60:1, 1.65:1, 1.70:1, 1.75:1, 1.80:1, 1.85:1, or 1.90:1; enrich.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising a CXCR2 agonist and a CXCR4 antagonist. , after administration of the CXCR2 agonist and the CXCR4 antagonist, the frequency of CD34 ⁺ cells in a sample of peripheral blood of the donor is about 0.051% to about 0.140.
% of the population of cells. In some embodiments, the cell population has a frequency of CD34 ⁺ cells of about 0.051%, 0.052%, 0.053%, 0
.054%, 0.055%, 0.056%, 0.057%, 0.058%, 0.059%, 0.060%, 0.061%, 0.062%, 0.063%, 0
.064%, 0.065%, 0.066%, 0.067%, 0.068%, 0.069%, 0.070%, 0.071%, 0.072%, 0.073%, 0
.074%, 0.075%, 0.076%, 0.077%, 0.078%, 0.079%, 0.080%, 0.081%, 0.082%, 0.083%, 0
.084%, 0.085%, 0.086%, 0.087%, 0.088%, 0.089%, 0.090%, 0.091%, 0.092%, 0.093%, 0
.094%, 0.095%, 0.096%, 0.097%, 0.098%, 0.099%, 0.100%, 0.101%, 0.102%, 0.103%, 0
.104%, 0.105%, 0.106%, 0.107%, 0.108%, 0.109%, 0.110%, 0.111%, 0.112%, 0.113%, 0
.114%, 0.115%, 0.116%, 0.117%, 0.118%, 0.119%, 0.120%, 0.121%, 0.122%, 0.123%, 0
.124%, 0.125%, 0.126%, 0.127%, 0.128%, 0.129%, 0.130%, 0.131%, 0.132%, 0.133%, 0
May be .134%, 0.135%, 0.136%, 0.137%, 0.138%, 0.139%, or 0.140%. In some embodiments, the population of cells has a frequency of CD34 ⁺ cells of about 0.050% to about 0.120%, about 0.0
60% to about 0.110%, or about 0.080% to about 0.100%. In some embodiments, the population of cells has a frequency of CD34 ⁺ cells of about 0.080% to about 0.120%, e.g., about 0.080%, 0.081%, 0.082%, 0
.083%, 0.084%, 0.085%, 0.086%, 0.087%, 0.088%, 0.089%, 0.090%, 0.091%, 0.092%, 0
.093%, 0.094%, 0.095%, 0.096%, 0.097%, 0.098%, 0.099%, 0.100%, 0.101%, 0.102%, 0
.103%, 0.104%, 0.105%, 0.106%, 0.107%, 0.108%, 0.109%, 0.110%, 0.111%, 0.112%, 0
Hematopoietic stem cell frequency of .113%, 0.114%, 0.115%, 0.116%, 0.117%, 0.118%, 0.119%, or 0.120%. In some embodiments, the population of cells has a frequency of CD34 ⁺ cells of about 0.097%.
It is.

In a further aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising
Agonists and CXCR4 antagonists are evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and the CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and the CXCR4 antagonist. the frequency of CD34 ⁺ cells in the donor's peripheral blood is at least 3 times greater (e.g., about 3.4 times to about 7.1 times, such as about 3.4 times, 3.5 times, 3.6 times, 3.7 times, 3.8 times, 3.9 times) times, 4.0 times, 4.1 times, 4.2
times, 4.3 times, 4.4 times, 4.5 times, 4.6 times, 4.7 times, 4.8 times, 4.9 times, 5.0 times, 5.1 times, 5.2 times, 5.3 times,
5.4x, 5.5x, 5.6x, 5.7x, 5.8x, 5.9x, 6.0x, 6.1x, 6.2x, 6.3x, 6.4x, 6.
5-fold, 6.6-fold, 6.7-fold, 6.8-fold, 6.9-fold, 7.0-fold, or 7.1-fold) to said donor. In some embodiments, after administration of the CXCR2 agonist and CXCR4 antagonist, the frequency of CD34 ⁺ cells in the donor's peripheral blood increases by about 4-fold to about 7-fold, about 4
.5 times to about 6.5 times, or about 5 times to about 6 times. In some embodiments, after administration of the CXCR2 agonist and CXCR4 antagonist, the frequency of CD34 ⁺ cells in the donor's peripheral blood is about
4.0 times to approximately 6.0 times, for example, approximately 4.0 times, 4.1 times, 4.2 times, 4.3 times, 4.4 times, 4.5 times, 4.6 times, 4.7 times, 4.8 times, 4.9 times, 5.0 times, 5.1 times, 5.2 times, 5.3 times, 5.4 times, 5.5 times, 5.6 times, 5.7 times, 5.8 times, 5
.9 times or 6.0 times. In some embodiments, the frequency of CD34 ⁺ cells in peripheral blood of said donor increases about 4.8-fold after administration of a CXCR2 agonist and a CXCR4 antagonist.

In a further embodiment, the invention provides methods for obtaining bone marrow from a mammalian donor (e.g., a human donor).
The method relates to a method of mobilizing a population of hematopoietic stem cells in peripheral blood, the method comprising CXCR2
agonist and CXCR4 antagonist, said CXCR2 agonist and said CXCR4 antagonist.
CD34 on leukocytes in a sample of the donor's peripheral blood after administration of⁺CD90⁺ CD45RA^-
said dosing in an amount sufficient to produce a population of cells having a ratio of about 0.0003 to about 0.0016 cells.
This includes administering to the patient. In some embodiments, for leukocytes in the sample
CD34⁺CD90⁺ CD45RA^-The cell ratio is approximately 0.00030, 0.00031, 0.00032, 0.00033, 0.00034,
0.00035, 0.00036, 0.00037, 0.00038, 0.00039, 0.00040, 0.00041, 0.00042, 0.00043,
0.00044, 0.00045, 0.00046, 0.00047, 0.00048, 0.00049, 0.00050, 0.00051, 0.00052
, 0.00053, 0.00054, 0.00055, 0.00056, 0.00057, 0.00058, 0.00059, 0.00060, 0.0006
1, 0.00062, 0.00063, 0.00064, 0.00065, 0.00066, 0.00067, 0.00068, 0.00069, 0.000
70, 0.00071, 0.00072, 0.00073, 0.00074, 0.00075, 0.00076, 0.00077, 0.00078, 0.00
079, 0.00080, 0.00081, 0.00082, 0.00083, 0.00084, 0.00085, 0.00086, 0.00087, 0.0
0088, 0.00089, 0.00090, 0.00091, 0.00092, 0.00093, 0.00094, 0.00095, 0.00096, 0.
00097, 0.00098, 0.00099, 0.00100, 0.00101, 0.00102, 0.00103, 0.00104, 0.00105, 0
.00106, 0.00107, 0.00108, 0.00109, 0.00110, 0.00111, 0.00112, 0.00113, 0.00114,
0.00115, 0.00116, 0.00117, 0.00118, 0.00119, 0.00120, 0.00121, 0.00122, 0.00123,
0.00124, 0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00132
, 0.00133, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.0014
1, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.001
50, 0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158, 0.00
159, or 0.00160. In some embodiments, leukemia in said sample
CD34 against the ball⁺CD90⁺ CD45RA^-The cell ratio is about 0.0008 to about 0.0010. Several
In embodiments, CD34 on leukocytes in said sample⁺CD90⁺ CD45RA^-The proportion of cells is approximately 0
.0006 to about 0.0012, for example, about 0.00060, 0.00061, 0.00062, 0.00063, 0.00064, 0.00065,
0.00066, 0.00067, 0.00068, 0.00069, 0.00070, 0.00071, 0.00072, 0.00073, 0.00074
, 0.00075, 0.00076, 0.00077, 0.00078, 0.00079, 0.00080, 0.00081, 0.00082, 0.0008
3, 0.00084, 0.00085, 0.00086, 0.00087, 0.00088, 0.00089, 0.00090, 0.00091, 0.000
92, 0.00093, 0.00094, 0.00095, 0.00096, 0.00097, 0.00098, 0.00099, 0.00100, 0.00
101, 0.00102, 0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108, 0.00109, 0.0
0110, 0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117, 0.00118, 0.
00119, or 0.00120, of CD34 on leukocytes in said sample⁺ CD90⁺CD45RA^-cell ratio
rate. In some embodiments, CD34 on leukocytes in said sample⁺CD90⁺ CD4
5RA^-The cell ratio is approximately 0.0009.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising a CXCR2 agonist and a CXCR4 antagonist. , when evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and the CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and the CXCR4 antagonist, about administering to said donor an amount sufficient to enrich said donor's peripheral blood with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to white blood cells at a ratio of 5.5:1 to about 26.9:1. In some embodiments, the peripheral blood of the donor is about 5.50:1, 5.55:1, 5.60:1, 5.65:1, 5.70:1,
5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1, 6.15:1, 6.20:1,
6.25:1, 6.30:1, 6.35:1, 6.40:1, 6.45:1, 6.50:1, 6.55:1, 6.60:1, 6.65:1, 6.70:1,
6.75:1, 6.80:1, 6.85:1, 6.90:1, 6.95:1, 7.00:1, 7.05:1, 7.10:1, 7.15:1, 7.20:1,
7.25:1, 7.30:1, 7.35:1, 7.40:1, 7.45:1, 7.50:1, 7.55:1, 7.60:1, 7.65:1, 7.70:1,
7.75:1, 7.80:1, 7.85:1, 7.90:1, 7.95:1, 8.00:1, 8.05:1, 8.10:1, 8.15:1, 8.20:1,
8.25:1, 8.30:1, 8.35:1, 8.40:1, 8.45:1, 8.50:1, 8.55:1, 8.60:1, 8.65:1, 8.70:1,
8.75:1, 8.80:1, 8.85:1, 8.90:1, 8.95:1, 9.00:1, 9.05:1, 9.10:1, 9.15:1, 9.20:1,
9.25:1, 9.30:1, 9.35:1, 9.40:1, 9.45:1, 9.50:1, 9.55:1, 9.60:1, 9.65:1, 9.70:1,
9.75:1, 9.80:1, 9.85:1, 9.90:1, 9.95:1, 10.00:1, 10.05:1, 10.10:1, 10.15:1, 10.2
0:1, 10.25:1, 10.30:1, 10.35:1, 10.40:1, 10.45:1, 10.50:1, 10.55:1, 10.60:1, 10.
65:1, 10.70:1, 10.75:1, 10.80:1, 10.85:1, 10.90:1, 10.95:1, 11.00:1, 11.05:1, 11
.10:1, 11.15:1, 11.20:1, 11.25:1, 11.30:1, 11.35:1, 11.40:1, 11.45:1, 11.50:1, 1
1.55:1, 11.60:1, 11.65:1, 11.70:1, 11.75:1, 11.80:1, 11.85:1, 11.90:1, 11.95:1,
12.00:1, 12.05:1, 12.10:1, 12.15:1, 12.20:1, 12.25:1, 12.30:1, 12.35:1, 12.40:1,
12.45:1, 12.50:1, 12.55:1, 12.60:1, 12.65:1, 12.70:1, 12.75:1, 12.80:1, 12.85:1
, 12.90:1, 12.95:1, 13.00:1, 13.05:1, 13.10:1, 13.15:1, 13.20:1, 13.25:1, 13.30:
1, 13.35:1, 13.40:1, 13.45:1, 13.50:1, 13.55:1, 13.60:1, 13.65:1, 13.70:1, 13.75
:1, 13.80:1, 13.85:1, 13.90:1, 13.95:1, 14.00:1, 14.05:1, 14.10:1, 14.15:1, 14.2
0:1, 14.25:1, 14.30:1, 14.35:1, 14.40:1, 14.45:1, 14.50:1, 14.55:1, 14.60:1, 14.
65:1, 14.70:1, 14.75:1, 14.80:1, 14.85:1, 14.90:1, 14.95:1, 15.00:1, 15.05:1, 15
.10:1, 15.15:1, 15.20:1, 15.25:1, 15.30:1, 15.35:1, 15.40:1, 15.45:1, 15.50:1, 1
5.55:1, 15.60:1, 15.65:1, 15.70:1, 15.75:1, 15.80:1, 15.85:1, 15.90:1, 15.95:1,
16.00:1, 16.05:1, 16.10:1, 16.15:1, 16.20:1, 16.25:1, 16.30:1, 16.35:1, 16.40:1,
16.45:1, 16.50:1, 16.55:1, 16.60:1, 16.65:1, 16.70:1, 16.75:1, 16.80:1, 16.85:1
, 16.90:1, 16.95:1, 17.00:1, 17.05:1, 17.10:1, 17.15:1, 17.20:1, 17.25:1, 17.30:
1, 17.35:1, 17.40:1, 17.45:1, 17.50:1, 17.55:1, 17.60:1, 17.65:1, 17.70:1, 17.75
:1, 17.80:1, 17.85:1, 17.90:1, 17.95:1, 18.00:1, 18.05:1, 18.10:1, 18.15:1, 18.2
0:1, 18.25:1, 18.30:1, 18.35:1, 18.40:1, 18.45:1, 18.50:1, 18.55:1, 18.60:1, 18.
65:1, 18.70:1, 18.75:1, 18.80:1, 18.85:1, 18.90:1, 18.95:1, 19.00:1, 19.05:1, 19
.10:1, 19.15:1, 19.20:1, 19.25:1, 19.30:1, 19.35:1, 19.40:1, 19.45:1, 19.50:1, 1
9.55:1, 19.60:1, 19.65:1, 19.70:1, 19.75:1, 19.80:1, 19.85:1, 19.90:1, 19.95:1,
20.00:1, 20.05:1, 20.10:1, 20.15:1, 20.20:1, 20.25:1, 20.30:1, 20.35:1, 20.40:1,
20.45:1, 20.50:1, 20.55:1, 20.60:1, 20.65:1, 20.70:1, 20.75:1, 20.80:1, 20.85:1
, 20.90:1, 20.95:1, 21.00:1, 21.05:1, 21.10:1, 21.15:1, 21.20:1, 21.25:1, 21.30:
1, 21.35:1, 21.40:1, 21.45:1, 21.50:1, 21.55:1, 21.60:1, 21.65:1, 21.70:1, 21.75
:1, 21.80:1, 21.85:1, 21.90:1, 21.95:1, 22.00:1, 22.05:1, 22.10:1, 22.15:1, 22.2
0:1, 22.25:1, 22.30:1, 22.35:1, 22.40:1, 22.45:1, 22.50:1, 22.55:1, 22.60:1, 22.
65:1, 22.70:1, 22.75:1, 22.80:1, 22.85:1, 22.90:1, 22.95:1, 23.00, 23.05:1, 23.1
0:1, 23.15:1, 23.20:1, 23.25:1, 23.30:1, 23.35:1, 23.40:1, 23.45:1, 23.50:1, 23.
55:1, 23.60:1, 23.65:1, 23.70:1, 23.75:1, 23.80:1, 23.85:1, 23.90:1, 23.95:1, 24
.00:1, 24.05:1, 24.10:1, 24.15:1, 24.20:1, 24.25:1, 24.30:1, 24.35:1, 24.40:1, 2
4.45:1, 24.50:1, 24.55:1, 24.60:1, 24.65:1, 24.70:1, 24.75:1, 24.80:1, 24.85:1,
24.90:1, 24.95:1, 25.05:1, 25.10:1, 25.15:1, 25.20:1, 25.25:1, 25.30:1, 25.35:1,
25.40:1, 25.45:1, 25.50:1, 25.55:1, 25.60:1, 25.65:1, 25.70:1, 25.75:1, 25.80:1
, 25.85:1, 25.90:1, 25.95:1, 26.00:1, 26.05:1, 26.10:1, 26.15:1, 26.20:1, 26.25:
1, 26.30:1, 26.35:1, 26.40:1, 26.45:1, 26.50:1, 26.55:1, 26.60:1, 26.65:1, 26.70
:1, 26.75:1, 26.80:1, 26.85:1, 26.90:1, or 26.95:1 ratio of CD34 to white blood cells.
⁺ CD90 ⁺ CD45RA ^- cells may be enriched. In some embodiments, the peripheral blood of the donor is in a ratio of about 5.5:1 to about 6.5:1, e.g., about 5.50:1, 5.55:1, 5.60:1, 5.65:1,
5.70:1, 5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1, 6.15:1,
Enrich with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to leukocytes to a ratio of 6.20:1, 6.25:1, 6.30:1, 6.35:1, 6.40:1, 6.45:1, or 6.50:1. In some embodiments, the donor's peripheral blood is enriched with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to leukocytes at a ratio of about 6.0:1.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising at least about
Density of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells of 16,000 cells/ml, e.g., about 20,000 cells/ml to about 75,000
cells/ml, about 25,000 cells/ml to about 70,000 cells/ml, about 30,000 cells/ml to about 65,000 cells/ml, about 3
5,000 cells/ml to about 60,000 cells/ml, about 40,000 cells/ml to about 55,000 cells/ml, or about 45,000 cells/ml to about 50,000 cells/ml (e.g., about 16,000 cells/ml, 17,000 cells/ml, 18,000 cells/ml cells/ml, 1
9,000 cells/ml, 20,000 cells/ml, 21,000 cells/ml, 22,000 cells/ml, 23,000 cells/ml, 24,000
cells/ml, 25,000 cells/ml, 26,000 cells/ml, 27,000 cells/ml, 28,000 cells/ml, 29,000 cells/
ml, 30,000 cells/ml, 31,000 cells/ml, 32,000 cells/ml, 33,000 cells/ml, 34,000 cells/ml, 3
5,000 cells/ml, 36,000 cells/ml, 37,000 cells/ml, 38,000 cells/ml, 39,000 cells/ml, 40,000
cells/ml, 41,000 cells/ml, 42,000 cells/ml, 43,000 cells/ml, 44,000 cells/ml, 45,000 cells/
ml, 46,000 cells/ml, 47,000 cells/ml, 48,000 cells/ml, 49,000 cells/ml, 50,000 cells/ml, 5
1,000 cells/ml, 52,000 cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000
cells/ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000 cells/ml, 61,000 cells/
ml, 62,000 cells/ml, 63,000 cells/ml, 64,000 cells/ml, 65,000 cells/ml, 66,000 cells/ml, 6
7,000 cells/ml, 68,000 cells/ml, 69,000 cells/ml, 70,000 cells/ml, 71,000 cells/ml, 72,000
cells/ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000 cells/ml, 77,000 cells/
ml, or more) with a density of CD34 ⁺ CD90 ⁺ CD45RA ^- cells, and approximately 5.3 x 10 ⁷ cells/m
leukocyte density of less than l, e.g. about 2.3 x 10 ⁷ cells/ml to about 5.3 x 10 ⁷ cells/ml, about 2.5 x 10 ⁷
cells/ml ~ approx. 5.1 x 10 ⁷ cells/ml, 2.9 x 10 ⁷ cells/ml ~ approx. 4.5 x 10 ⁷ cells/ml, approx. 3 x 10 ⁷ cells
/ml to about 4 x 10 ⁷ cells/ml (e.g. 5.3 x 10 ⁷ cells/ml, 5.2 x 10 ⁷ cells/ml, 5.1 x 10 ⁷ cells
/ml, 5 x 10 ⁷ cells/ml, 4.9 x 10 ⁷ cells/ml, 4.8 x 10 ⁷ cells/ml, 4.7 x 10 ⁷ cells/ml, 4.6 x
10 ⁷ cells/ml, 4.5 x 10 ⁷ cells/ml, 4.4 x 10 ⁷ cells/ml, 4.3 x 10 ⁷ cells/ml 4.2 x 10 ⁷ cells/ml
ml, 4.1 x 10 ⁷ cells/ml 4 x 10 ⁷ cells/ml, 3.9 x 10 ⁷ cells/ml, 3.8 x 10 ⁷ cells/ml, 3.7 x 1
0 ⁷ cells/ml, 3.6 x 10 ⁷ cells/ml, 3.5 x 10 ⁷ cells/ml, 3.4 x 10 ⁷ cells/ml, 3.3 x 10 ⁷ cells/m
l, 3.2 x 10 ⁷ cells/ml, 3.1 x 10 ⁷ cells/ml, 3 x 10 ⁷ cells/ml, 2.9 x 10 ⁷ cells/ml, 2.8 x 1
0 ⁷ cells/ml, 2.7 x 10 ⁷ cells/ml, 2.6 x 10 ⁷ cells/ml, 2.5 x 10 ⁷ cells/ml, 2.4 x 10 ⁷ cells/m
administering to said donor a CXCR2 agonist and a CXCR4 antagonist in an amount sufficient to produce a population of cells having a density of white blood cells of 1, 2.3 x 10 ⁷ cells/ml, or less). In some embodiments, the method provides between about 20,000 cells/ml and about 75,000 cells/ml of CD34
^{CXCR2 agonist and _ _} administering a CXCR4 antagonist to said donor. In some embodiments, the method comprises about
having a density of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells of 30,000 cells/ml to about 60,000 cells/ml, and about 2.5
administering to said donor a CXCR2 agonist and a CXCR4 antagonist in an amount sufficient to produce a cell population having a density of white blood cells from x10 ⁷ cells/ml to about 5 x 10 ⁷ cells/ml. In some embodiments, the method provides between about 40,000 cells/ml and about 50,000 cells/ml of CD34 ⁺ CD90 ⁺ CD4
5RA ^- a CXCR2 agonist and a CXCR4 antagonist in an amount sufficient to produce a cell population having a density of 5RA - cells and a density of white blood cells of about 3 x 10 ⁷ cells/ml to about 4 x 10 ⁷ cells/ml. and administering to said donor.

In a further embodiment, the invention provides methods for obtaining bone marrow from a mammalian donor (e.g., a human donor).
The method relates to a method for mobilizing a population of hematopoietic stem cells in peripheral blood, the method comprising CXCR2
agonist and CXCR4 antagonist, said CXCR2 agonist and said CXCR4 antagonist.
CD34 on neutrophils in a sample of the donor's peripheral blood after administration of⁺CD90⁺ CD45RA^-
said dosing in an amount sufficient to produce a population of cells having a ratio of about 0.0007 to about 0.0043 cells.
This includes administering to the patient. In some embodiments, for neutrophils in the sample
CD34⁺CD90⁺ CD45RA^-The cell ratio is approximately 0.00070, 0.00071, 0.00072, 0.00073, 0.00074,
0.00075, 0.00076, 0.00077, 0.00078, 0.00079, 0.00080, 0.00081, 0.00082, 0.00083,
0.00084, 0.00085, 0.00086, 0.00087, 0.00088, 0.00089, 0.00090, 0.00091, 0.00092
, 0.00093, 0.00094, 0.00095, 0.00096, 0.00097, 0.00098, 0.00099, 0.00100, 0.0010
1, 0.00102, 0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108, 0.00109, 0.001
10, 0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117, 0.00118, 0.00
119, 0.00120, 0.00121, 0.00122, 0.00123, 0.00124, 0.00125, 0.00126, 0.00127, 0.0
0128, 0.00129, 0.00130, 0.00131, 0.00132, 0.00133, 0.00134, 0.00135, 0.00136, 0.
00137, 0.00138, 0.00139, 0.00140, 0.00141, 0.00142, 0.00143, 0.00144, 0.00145, 0
.00146, 0.00147, 0.00148, 0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154,
0.00155, 0.00156, 0.00157, 0.00158, 0.00159, 0.00160, 0.00161, 0.00162, 0.00163,
0.00164, 0.00165, 0.00166, 0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172
, 0.00173, 0.00174, 0.00175, 0.00176, 0.00177, 0.00178, 0.00179, 0.00180, 0.0018
1, 0.00182, 0.00183, 0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189, 0.001
90, 0.00191, 0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197, 0.00198, 0.00
199, 0.00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205, 0.00206, 0.00207, 0.0
0208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00213, 0.00214, 0.00215, 0.00216, 0.
00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.00222, 0.00223, 0.00224, 0.00225, 0
.00226, 0.00227, 0.00228, 0.00229, 0.00230, 0.00231, 0.00232, 0.00233, 0.00234,
0.00235, 0.00236, 0.00237, 0.00238, 0.00239, 0.00240, 0.00241, 0.00242, 0.00243,
0.00244, 0.00245, 0.00246, 0.00247, 0.00248, 0.00249, 0.00250, 0.00251, 0.00252
, 0.00253, 0.00254, 0.00255, 0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.0026
1, 0.00262, 0.00263, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.002
70, 0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00277, 0.00278, 0.00
279, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.0
0288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.
00297, 0.00298, 0.00299, 0.00300, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0
.00305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313,
0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322,
0.00323, 0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331
, 0.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339, 0.0034
0, 0.00341, 0.00342, 0.00343, 0.00344, 0.00345, 0.00346, 0.00347, 0.00348, 0.003
49, 0.00350, 0.00351, 0.00352, 0.00353, 0.00354, 0.00355, 0.00356, 0.00357, 0.00
358, 0.00359, 0.00360, 0.00361, 0.00362, 0.00363, 0.00364, 0.00365, 0.00366, 0.0
0367, 0.00368, 0.00369, 0.00370, 0.00371, 0.00372, 0.00373, 0.00374, 0.00375, 0.
00376, 0.00377, 0.00378, 0.00379, 0.00380, 0.00381, 0.00382, 0.00383, 0.00384, 0
.00385, 0.00386, 0.00387, 0.00388, 0.00389, 0.00390, 0.00391, 0.00392, 0.00393,
0.00394, 0.00395, 0.00396, 0.00397, 0.00398, 0.00399, 0.00400, 0.00401, 0.00402,
0.00403, 0.00404, 0.00405, 0.00406, 0.00407, 0.00408, 0.00409, 0.00410, 0.00411
, 0.00412, 0.00413, 0.00414, 0.00415, 0.00416, 0.00417, 0.00418, 0.00419, 0.0042
0, 0.00421, 0.00422, 0.00423, 0.00424, 0.00425, 0.00426, 0.00427, 0.00428, 0.004
29, or 0.00430. In some embodiments, neutrophils in said sample
CD34 against⁺CD90⁺ CD45RA^-The cell ratio is about 0.002 to about 0.003. some implementation
In terms of morphology, CD34 on neutrophils in the sample⁺CD90⁺ CD45RA^-The cell ratio is approximately 0.001
4 to about 0.0034, e.g., about 0.00140, 0.00141, 0.00142, 0.00143, 0.00144, 0.00145, 0.0
0146, 0.00147, 0.00148, 0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154, 0.
00155, 0.00156, 0.00157, 0.00158, 0.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0
.00164, 0.00165, 0.00166, 0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172,
0.00173, 0.00174, 0.00175, 0.00176, 0.00177, 0.00178, 0.00179, 0.00180, 0.00181,
0.00182, 0.00183, 0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189, 0.00190
, 0.00191, 0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197, 0.00198, 0.0019
9, 0.00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205, 0.00206, 0.00207, 0.002
08, 0.00209, 0.00210, 0.00211, 0.00212, 0.00213, 0.00214, 0.00215, 0.00216, 0.00
217, 0.00218, 0.00219, 0.00220, 0.00221, 0.00222, 0.00223, 0.00224, 0.00225, 0.0
0226, 0.00227, 0.00228, 0.00229, 0.00230, 0.00231, 0.00232, 0.00233, 0.00234, 0.
00235, 0.00236, 0.00237, 0.00238, 0.00239, 0.00240, 0.00241, 0.00242, 0.00243, 0
.00244, 0.00245, 0.00246, 0.00247, 0.00248, 0.00249, 0.00250, 0.00251, 0.00252,
0.00253, 0.00254, 0.00255, 0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.00261,
0.00262, 0.00263, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270
, 0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00277, 0.00278, 0.0027
9, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.002
88, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.00
297, 0.00298, 0.00299, 0.00300, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.0
0305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313, 0.
00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0
.00323, 0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331,
0.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339, or 0.0
CD34 on neutrophils in said sample of 0340,⁺CD90⁺ CD45RA^-The ratio of cells is, stomach
In some embodiments, CD34 on neutrophils in said sample⁺CD90⁺ CD45RA^-cell ratio
The rate is approximately 0.0024.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising a CXCR2 agonist and a CXCR4 antagonist. , as assessed by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and the CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and the CXCR4 antagonist, about administering to said donor an amount sufficient to enrich said donor's peripheral blood with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to neutrophils in a ratio of 3.5:1 to about 22.0:1; include. In some embodiments, the peripheral blood of the donor is about 3.50:1, 3.55:1, 3.60:1, 3.65:1, 3.70:1,
3.75:1, 3.80:1, 3.85:1, 3.90:1, 3.95:1, 4.00:1, 4.05:1, 4.10:1, 4.15:1, 4.20:1,
4.25:1, 4.30:1, 4.35:1, 4.40:1, 4.45:1, 4.50:1, 4.55:1, 4.60:1, 4.65:1, 4.70:1,
4.75:1, 4.80:1, 4.85:1, 4.90:1, 4.95:1, 5.00:1, 5.05:1, 5.10:1, 5.15:1, 5.20:1,
5.25:1, 5.30:1, 5.35:1, 5.40:1, 5.45:1, 5.50:1, 5.55:1, 5.60:1, 5.65:1, 5.70:1,
5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1, 6.15:1, 6.20:1,
6.25:1, 6.30:1, 6.35:1, 6.40:1, 6.45:1, 6.50:1, 6.55:1, 6.60:1, 6.65:1, 6.70:1,
6.75:1, 6.80:1, 6.85:1, 6.90:1, 6.95:1, 7.00:1, 7.05:1, 7.10:1, 7.15:1, 7.20:1,
7.25:1, 7.30:1, 7.35:1, 7.40:1, 7.45:1, 7.50:1, 7.55:1, 7.60:1, 7.65:1, 7.70:1,
7.75:1, 7.80:1, 7.85:1, 7.90:1, 7.95:1, 8.00:1, 8.05:1, 8.10:1, 8.15:1, 8.20:1,
8.25:1, 8.30:1, 8.35:1, 8.40:1, 8.45:1, 8.50:1, 8.55:1, 8.60:1, 8.65:1, 8.70:1,
8.75:1, 8.80:1, 8.85:1, 8.90:1, 8.95:1, 9.00:1, 9.05:1, 9.10:1, 9.15:1, 9.20:1,
9.25:1, 9.30:1, 9.35:1, 9.40:1, 9.45:1, 9.50:1, 9.55:1, 9.60:1, 9.65:1, 9.70:1,
9.75:1, 9.80:1, 9.85:1, 9.90:1, 9.95:1, 10.00:1, 10.05:1, 10.10:1, 10.15:1, 10.2
0:1, 10.25:1, 10.30:1, 10.35:1, 10.40:1, 10.45:1, 10.50:1, 10.55:1, 10.60:1, 10.
65:1, 10.70:1, 10.75:1, 10.80:1, 10.85:1, 10.90:1, 10.95:1, 11.00:1, 11.05:1, 11
.10:1, 11.15:1, 11.20:1, 11.25:1, 11.30:1, 11.35:1, 11.40:1, 11.45:1, 11.50:1, 1
1.55:1, 11.60:1, 11.65:1, 11.70:1, 11.75:1, 11.80:1, 11.85:1, 11.90:1, 11.95:1,
12.00:1, 12.05:1, 12.10:1, 12.15:1, 12.20:1, 12.25:1, 12.30:1, 12.35:1, 12.40:1,
12.45:1, 12.50:1, 12.55:1, 12.60:1, 12.65:1, 12.70:1, 12.75:1, 12.80:1, 12.85:1
, 12.90:1, 12.95:1, 13.00:1, 13.05:1, 13.10:1, 13.15:1, 13.20:1, 13.25:1, 13.30:
1, 13.35:1, 13.40:1, 13.45:1, 13.50:1, 13.55:1, 13.60:1, 13.65:1, 13.70:1, 13.75
:1, 13.80:1, 13.85:1, 13.90:1, 13.95:1, 14.00:1, 14.05:1, 14.10:1, 14.15:1, 14.2
0:1, 14.25:1, 14.30:1, 14.35:1, 14.40:1, 14.45:1, 14.50:1, 14.55:1, 14.60:1, 14.
65:1, 14.70:1, 14.75:1, 14.80:1, 14.85:1, 14.90:1, 14.95:1, 15.00:1, 15.05:1, 15
.10:1, 15.15:1, 15.20:1, 15.25:1, 15.30:1, 15.35:1, 15.40:1, 15.45:1, 15.50:1, 1
5.55:1, 15.60:1, 15.65:1, 15.70:1, 15.75:1, 15.80:1, 15.85:1, 15.90:1, 15.95:1,
16.00:1, 16.05:1, 16.10:1, 16.15:1, 16.20:1, 16.25:1, 16.30:1, 16.35:1, 16.40:1,
16.45:1, 16.50:1, 16.55:1, 16.60:1, 16.65:1, 16.70:1, 16.75:1, 16.80:1, 16.85:1
, 16.90:1, 16.95:1, 17.00:1, 17.05:1, 17.10:1, 17.15:1, 17.20:1, 17.25:1, 17.30:
1, 17.35:1, 17.40:1, 17.45:1, 17.50:1, 17.55:1, 17.60:1, 17.65:1, 17.70:1, 17.75
:1, 17.80:1, 17.85:1, 17.90:1, 17.95:1, 18.00:1, 18.05:1, 18.10:1, 18.15:1, 18.2
0:1, 18.25:1, 18.30:1, 18.35:1, 18.40:1, 18.45:1, 18.50:1, 18.55:1, 18.60:1, 18.
65:1, 18.70:1, 18.75:1, 18.80:1, 18.85:1, 18.90:1, 18.95:1, 19.00:1, 19.05:1, 19
.10:1, 19.15:1, 19.20:1, 19.25:1, 19.30:1, 19.35:1, 19.40:1, 19.45:1, 19.50:1, 1
9.55:1, 19.60:1, 19.65:1, 19.70:1, 19.75:1, 19.80:1, 19.85:1, 19.90:1, 19.95:1,
20.00:1, 20.05:1, 20.10:1, 20.15:1, 20.20:1, 20.25:1, 20.30:1, 20.35:1, 20.40:1,
20.45:1, 20.50:1, 20.55:1, 20.60:1, 20.65:1, 20.70:1, 20.75:1, 20.80:1, 20.85:1
, 20.90:1, 20.95:1, 21.00:1, 21.05:1, 21.10:1, 21.15:1, 21.20:1, 21.25:1, 21.30:
1, 21.35:1, 21.40:1, 21.45:1, 21.50:1, 21.55:1, 21.60:1, 21.65:1, 21.70:1, 21.75
:1, 21.80:1, 21.85:1, 21.90:1, 21.95:1, or 22.00:1 ratio of CD34 to neutrophils.
⁺ CD90 ⁺ CD45RA ^- cells may be enriched. In some embodiments, the donor's peripheral blood is enriched with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to neutrophils at a ratio of about 7.0:1 to about 10:1. In some embodiments, the peripheral blood of the donor is in a ratio of about 7.00:1 to about 9.00:1, e.g., about 7.00:1, 7.05:1, 7.10:1, 7.15:1, 7.20:1, 7.25:1, 7.30:1, 7.35:1, 7.
40:1, 7.45:1, 7.50:1, 7.55:1, 7.60:1, 7.65:1, 7.70:1, 7.75:1, 7.80:1, 7.85:1, 7.
90:1, 7.95:1, 8.00:1, 8.05:1, 8.10:1, 8.15:1, 8.20:1, 8.25:1, 8.30:1, 8.35:1, 8.
40:1, 8.45:1, 8.50:1, 8.55:1, 8.60:1, 8.65:1, 8.70:1, 8.75:1, 8.80:1, 8.85:1, 8.
Enrich with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to neutrophils to a ratio of 90:1, 8.95:1, or 9.00:1. In some embodiments, the donor's peripheral blood is enriched with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to neutrophils at a ratio of about 8.2:1.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising at least about
Density of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells of 16,000 cells/ml, e.g., about 20,000 cells/ml to about 75,000
cells/ml, about 25,000 cells/ml to about 70,000 cells/ml, about 30,000 cells/ml to about 65,000 cells/ml, about 3
5,000 cells/ml to about 60,000 cells/ml, about 40,000 cells/ml to about 55,000 cells/ml, or about 45,000 cells/ml to about 50,000 cells/ml (e.g., about 16,000 cells/ml, 17,000 cells/ml, 18,000 cells/ml cells/ml, 1
9,000 cells/ml, 20,000 cells/ml, 21,000 cells/ml, 22,000 cells/ml, 23,000 cells/ml, 24,000
cells/ml, 25,000 cells/ml, 26,000 cells/ml, 27,000 cells/ml, 28,000 cells/ml, 29,000 cells/
ml, 30,000 cells/ml, 31,000 cells/ml, 32,000 cells/ml, 33,000 cells/ml, 34,000 cells/ml, 3
5,000 cells/ml, 36,000 cells/ml, 37,000 cells/ml, 38,000 cells/ml, 39,000 cells/ml, 40,000
cells/ml, 41,000 cells/ml, 42,000 cells/ml, 43,000 cells/ml, 44,000 cells/ml, 45,000 cells/
ml, 46,000 cells/ml, 47,000 cells/ml, 48,000 cells/ml, 49,000 cells/ml, 50,000 cells/ml, 5
1,000 cells/ml, 52,000 cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000
cells/ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000 cells/ml, 61,000 cells/
ml, 62,000 cells/ml, 63,000 cells/ml, 64,000 cells/ml, 65,000 cells/ml, 66,000 cells/ml, 6
7,000 cells/ml, 68,000 cells/ml, 69,000 cells/ml, 70,000 cells/ml, 71,000 cells/ml, 72,000
cells/ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000 cells/ml, 77,000 cells/
ml, or more) and have a density of CD34 ⁺ CD90 ⁺ CD45RA ^- cells of approximately 2.5 x 10 ⁷ cells/m
a density of neutrophils of less than l, e.g., about 1 x 10 ⁷ cells/ml to about 2.5 x 10 ⁷ cells/ml, about 1.3 x 10 ⁷ cells/ml to about 2 x 10 ⁷ cells/ml, or about 1.5 x 10 ⁷ cells/ml to about 1.9 x 10 ⁷ cells/ml (e.g. about 2
.5 x 10 ⁷ cells/ml, 2.4 x 10 ⁷ cells/ml, 2.3 x 10 ⁷ cells/ml, 2.2 x 0 ⁷ cells/ml, 2.1 x 10 ⁷
cells/ml, 2 x 10 ⁷ cells/ml, 1.9 x 10 ⁷ cells/ml, 1.8 x 10 ⁷ cells/ml, 1.7 x 10 ⁷ cells/ml, 1
.6 x 10 ⁷ cells/ml, 1.5 x 10 ⁷ cells/ml 1.4 x 10 ⁷ cells/ml, 1.3 x 10 ⁷ cells/ml, 1.2 x 10 ⁷
CXCR2 agonist and CXCR4 in an amount sufficient to produce a population of cells having a density of neutrophils of 1.1 x 10 ⁷ cells/ml, 1 x 10 ⁷ cells/ml, or less). administering an antagonist to said donor. In some embodiments, the method uses about 20,000 cells/ml.
~ with a density of CD34 ⁺ CD90 ⁺ CD45RA ^- cells of ~75,000 cells/ml, and ~1 x 10 ⁷ cells/ml ~
administering to said donor a CXCR2 agonist and a CXCR4 antagonist in an amount sufficient to produce a cell population having a density of neutrophils of about 2.5×10 ⁷ cells/ml. In some embodiments, the method has a density of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells from about 30,000 cells/ml to about 60,000 cells/ml, and from about 1.3×10 ⁷ cells/ml to about 2.3×10 ⁷ administering to said donor a CXCR2 agonist and a CXCR4 antagonist in amounts sufficient to produce a population of cells having a density of neutrophils/ml. In some embodiments, the method provides between about 40,000 cells/ml and about 50,00
with a density of CD34 ⁺ CD90 ⁺ CD45RA ^- cells of 0 cells/ml, and from about 1.5 x 10 ⁷ cells/ml to about 2 x 10 ⁷
administering to said donor a CXCR2 agonist and a CXCR4 antagonist in amounts sufficient to produce a population of cells having a density of neutrophils/ml.

In a further aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising
agonist and CXCR4 antagonist to CD34 ⁺ CD90 ⁺ CD45R lymphocytes in a sample of peripheral blood of said donor after administration of said CXCR2 agonist and said CXCR4 antagonist.
administering to said donor an amount sufficient to produce a population of cells in which the proportion of A ^- cells is from about 0.0008 to about 0.0069. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to lymphocytes in the sample is about 0.00080, 0.00081, 0.00082, 0.00083, 0.000.
84, 0.00085, 0.00086, 0.00087, 0.00088, 0.00089, 0.00090, 0.00091, 0.00092, 0.00
093, 0.00094, 0.00095, 0.00096, 0.00097, 0.00098, 0.00099, 0.00100, 0.00101, 0.0
0102, 0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108, 0.00109, 0.00110, 0.
00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117, 0.00118, 0.00119, 0
.00120, 0.00121, 0.00122, 0.00123, 0.00124, 0.00125, 0.00126, 0.00127, 0.00128,
0.00129, 0.00130, 0.00131, 0.00132, 0.00133, 0.00134, 0.00135, 0.00136, 0.00137,
0.00138, 0.00139, 0.00140, 0.00141, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146
, 0.00147, 0.00148, 0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154, 0.0015
5, 0.00156, 0.00157, 0.00158, 0.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.001
64, 0.00165, 0.00166, 0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172, 0.00
173, 0.00174, 0.00175, 0.00176, 0.00178, 0.00179, 0.00180, 0.00181, 0.00182, 0.0
0183, 0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189, 0.00190, 0.00191, 0.
00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197, 0.00198, 0.00199, 0.00200, 0
.00201, 0.00202, 0.00203, 0.00204, 0.00205, 0.00206, 0.00207, 0.00208, 0.00209,
0.00210, 0.00211, 0.00212, 0.00213, 0.00214, 0.00215, 0.00216, 0.00217, 0.00218,
0.00219, 0.00220, 0.00221, 0.00222, 0.00223, 0.00224, 0.00225, 0.00226, 0.00227
, 0.00228, 0.00229, 0.00230, 0.00231, 0.00232, 0.00233, 0.00234, 0.00235, 0.0023
6, 0.00237, 0.00238, 0.00239, 0.00240, 0.00241, 0.00242, 0.00243, 0.00244, 0.002
45, 0.00246, 0.00247, 0.00248, 0.00249, 0.00250, 0.00251, 0.00252, 0.00253, 0.00
254, 0.00255, 0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.00261, 0.00262, 0.0
0263, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.
00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00278, 0.00279, 0.00280, 0.00281, 0
.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290,
0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.00297, 0.00298, 0.00299,
0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308
, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0.0031
7, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325, 0.003
26, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00
335, 0.00336, 0.00337, 0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343, 0.0
0344, 0.00345, 0.00346, 0.00347, 0.00348, 0.00349, 0.00350, 0.00351, 0.00352, 0.
00353, 0.00354, 0.00355, 0.00356, 0.00357, 0.00358, 0.00359, 0.00360, 0.00361, 0
.00362, 0.00363, 0.00364, 0.00365, 0.00366, 0.00367, 0.00368, 0.00369, 0.00370,
0.00371, 0.00372, 0.00373, 0.00374, 0.00375, 0.00376, 0.00378, 0.00379, 0.00380,
0.00381, 0.00382, 0.00383, 0.00384, 0.00385, 0.00386, 0.00387, 0.00388, 0.00389
, 0.00390, 0.00391, 0.00392, 0.00393, 0.00394, 0.00395, 0.00396, 0.00397, 0.0039
8, 0.00399, 0.00401, 0.00402, 0.00403, 0.00404, 0.00405, 0.00406, 0.00407, 0.004
08, 0.00409, 0.00410, 0.00411, 0.00412, 0.00413, 0.00414, 0.00415, 0.00416, 0.00
417, 0.00418, 0.00419, 0.00420, 0.00421, 0.00422, 0.00423, 0.00424, 0.00425, 0.0
0426, 0.00427, 0.00428, 0.00429, 0.00430, 0.00431, 0.00432, 0.00433, 0.00434, 0.
00435, 0.00436, 0.00437, 0.00438, 0.00439, 0.00440, 0.00441, 0.00442, 0.00443, 0
.00444, 0.00445, 0.00446, 0.00447, 0.00448, 0.00449, 0.00450, 0.00451, 0.00452,
0.00453, 0.00454, 0.00455, 0.00456, 0.00457, 0.00458, 0.00459, 0.00460, 0.00461,
0.00462, 0.00463, 0.00464, 0.00465, 0.00466, 0.00467, 0.00468, 0.00469, 0.00470
, 0.00471, 0.00472, 0.00473, 0.00474, 0.00475, 0.00476, 0.00478, 0.00479, 0.0048
0, 0.00481, 0.00482, 0.00483, 0.00484, 0.00485, 0.00486, 0.00487, 0.00488, 0.004
89, 0.00490, 0.00491, 0.00492, 0.00493, 0.00494, 0.00495, 0.00496, 0.00497, 0.00
498, 0.00499, 0.00500, 0.00501, 0.00502, 0.00503, 0.00504, 0.00505, 0.00506, 0.0
0507, 0.00508, 0.00509, 0.00510, 0.00511, 0.00512, 0.00513, 0.00514, 0.00515, 0.
00516, 0.00517, 0.00518, 0.00519, 0.00520, 0.00521, 0.00522, 0.00523, 0.00524, 0
.00525, 0.00526, 0.00527, 0.00528, 0.00529, 0.00530, 0.00531, 0.00532, 0.00533,
0.00534, 0.00535, 0.00536, 0.00537, 0.00538, 0.00539, 0.00540, 0.00541, 0.00542,
0.00543, 0.00544, 0.00545, 0.00546, 0.00547, 0.00548, 0.00549, 0.00550, 0.00551
, 0.00552, 0.00553, 0.00554, 0.00555, 0.00556, 0.00557, 0.00558, 0.00559, 0.0056
0, 0.00561, 0.00562, 0.00563, 0.00564, 0.00565, 0.00566, 0.00567, 0.00568, 0.005
69, 0.00570, 0.00571, 0.00572, 0.00573, 0.00574, 0.00575, 0.00576, 0.00578, 0.00
579, 0.00580, 0.00581, 0.00582, 0.00583, 0.00584, 0.00585, 0.00586, 0.00587, 0.0
0588, 0.00589, 0.00590, 0.00591, 0.00592, 0.00593, 0.00594, 0.00595, 0.00596, 0.
00597, 0.00598, 0.00599, 0.00600, 0.00601, 0.00602, 0.00603, 0.00604, 0.00605, 0
.00606, 0.00607, 0.00608, 0.00609, 0.00610, 0.00611, 0.00612, 0.00613, 0.00614,
0.00615, 0.00616, 0.00617, 0.00618, 0.00619, 0.00620, 0.00621, 0.00622, 0.00623,
0.00624, 0.00625, 0.00626, 0.00627, 0.00628, 0.00629, 0.00630, 0.00631, 0.00632
, 0.00633, 0.00634, 0.00635, 0.00636, 0.00637, 0.00638, 0.00639, 0.00640, 0.0064
1, 0.00642, 0.00643, 0.00644, 0.00645, 0.00646, 0.00647, 0.00648, 0.00649, 0.006
50, 0.00651, 0.00652, 0.00653, 0.00654, 0.00655, 0.00656, 0.00657, 0.00658, 0.00
659, 0.00660, 0.00661, 0.00662, 0.00663, 0.00664, 0.00665, 0.00666, 0.00667, 0.0
0668, 0.00669, 0.00670, 0.00671, 0.00672, 0.00673, 0.00674, 0.00675, 0.00676, 0.
00678, 0.00679, 0.00680, 0.00681, 0.00682, 0.00683, 0.00684, 0.00685, 0.00686, 0
May be .00687, 0.00688, 0.00689, or 0.00690. In some embodiments,
The ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to lymphocytes in the sample is about 0.0011 to about 0.0.
031, for example, approximately 0.00110, 0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.
00117, 0.00118, 0.00119, 0.00120, 0.00121, 0.00122, 0.00123, 0.00124, 0.00125, 0
.00126, 0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00132, 0.00133, 0.00134,
0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.00141, 0.00142, 0.00143,
0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.00150, 0.00151, 0.00152
, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158, 0.00159, 0.00160, 0.0016
1, 0.00162, 0.00163, 0.00164, 0.00165, 0.00166, 0.00167, 0.00168, 0.00169, 0.001
70, 0.00171, 0.00172, 0.00173, 0.00174, 0.00175, 0.00176, 0.00178, 0.00179, 0.00
180, 0.00181, 0.00182, 0.00183, 0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.0
0189, 0.00190, 0.00191, 0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197, 0.
00198, 0.00199, 0.00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205, 0.00206, 0
.00207, 0.00208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00213, 0.00214, 0.00215,
0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.00222, 0.00223, 0.00224,
0.00225, 0.00226, 0.00227, 0.00228, 0.00229, 0.00230, 0.00231, 0.00232, 0.00233
, 0.00234, 0.00235, 0.00236, 0.00237, 0.00238, 0.00239, 0.00240, 0.00241, 0.0024
2, 0.00243, 0.00244, 0.00245, 0.00246, 0.00247, 0.00248, 0.00249, 0.00250, 0.002
51, 0.00252, 0.00253, 0.00254, 0.00255, 0.00256, 0.00257, 0.00258, 0.00259, 0.00
260, 0.00261, 0.00262, 0.00263, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.0
0269, 0.00270, 0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00278, 0.
00279, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0
.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296,
0.00297, 0.00298, 0.00299, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305,
0.00306, 0.00307, 0.00308, 0.00309, or 0.00310. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to lymphocytes in the sample is about 0.0021.

In a further aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising
Agonists and CXCR4 antagonists are evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and the CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and the CXCR4 antagonist. said donor in an amount sufficient to enrich said donor's peripheral blood with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to lymphocytes at a ratio of about 5.6:1 to about 37.0:1. including administering. In some embodiments, the peripheral blood of the donor is about 5.60:1, 5.65:1, 5.70:1, 5.75:1, 5.80
:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1, 6.15:1, 6.20:1, 6.25:1, 6.30
:1, 6.35:1, 6.40:1, 6.45:1, 6.50:1, 6.55:1, 6.60:1, 6.65:1, 6.70:1, 6.75:1, 6.80
:1, 6.85:1, 6.90:1, 6.95:1, 7.00:1, 7.05:1, 7.10:1, 7.15:1, 7.20:1, 7.25:1, 7.30
:1, 7.35:1, 7.40:1, 7.45:1, 7.50:1, 7.55:1, 7.60:1, 7.65:1, 7.70:1, 7.75:1, 7.80
:1, 7.85:1, 7.90:1, 7.95:1, 8.00:1, 8.05:1, 8.10:1, 8.15:1, 8.20:1, 8.25:1, 8.30
:1, 8.35:1, 8.40:1, 8.45:1, 8.50:1, 8.55:1, 8.60:1, 8.65:1, 8.70:1, 8.75:1, 8.80
:1, 8.85:1, 8.90:1, 8.95:1, 9.00:1, 9.05:1, 9.10:1, 9.15:1, 9.20:1, 9.25:1, 9.30
:1, 9.35:1, 9.40:1, 9.45:1, 9.50:1, 9.55:1, 9.60:1, 9.65:1, 9.70:1, 9.75:1, 9.80
:1, 9.85:1, 9.90:1, 9.95:1, 10.00:1, 10.05:1, 10.10:1, 10.15:1, 10.20:1, 10.25:1
, 10.30:1, 10.35:1, 10.40:1, 10.45:1, 10.50:1, 10.55:1, 10.60:1, 10.65:1, 10.70:
1, 10.75:1, 10.80:1, 10.85:1, 10.90:1, 10.95:1, 11.00:1, 11.05:1, 11.10:1, 11.15
:1, 11.20:1, 11.25:1, 11.30:1, 11.35:1, 11.40:1, 11.45:1, 11.50:1, 11.55:1, 11.6
0:1, 11.65:1, 11.70:1, 11.75:1, 11.80:1, 11.85:1, 11.90:1, 11.95:1, 12.00:1, 12.
05:1, 12.10:1, 12.15:1, 12.20:1, 12.25:1, 12.30:1, 12.35:1, 12.40:1, 12.45:1, 12
.50:1, 12.55:1, 12.60:1, 12.65:1, 12.70:1, 12.75:1, 12.80:1, 12.85:1, 12.90:1, 1
2.95:1, 13.00:1, 13.05:1, 13.10:1, 13.15:1, 13.20:1, 13.25:1, 13.30:1, 13.35:1,
13.40:1, 13.45:1, 13.50:1, 13.55:1, 13.60:1, 13.65:1, 13.70:1, 13.75:1, 13.80:1,
13.85:1, 13.90:1, 13.95:1, 14.00:1, 14.05:1, 14.10:1, 14.15:1, 14.20:1, 14.25:1
, 14.30:1, 14.35:1, 14.40:1, 14.45:1, 14.50:1, 14.55:1, 14.60:1, 14.65:1, 14.70:
1, 14.75:1, 14.80:1, 14.85:1, 14.90:1, 14.95:1, 15.00:1, 15.05:1, 15.10:1, 15.15
:1, 15.20:1, 15.25:1, 15.30:1, 15.35:1, 15.40:1, 15.45:1, 15.50:1, 15.55:1, 15.6
0:1, 15.65:1, 15.70:1, 15.75:1, 15.80:1, 15.85:1, 15.90:1, 15.95:1, 16.00:1, 16.
05:1, 16.10:1, 16.15:1, 16.20:1, 16.25:1, 16.30:1, 16.35:1, 16.40:1, 16.45:1, 16
.50:1, 16.55:1, 16.60:1, 16.65:1, 16.70:1, 16.75:1, 16.80:1, 16.85:1, 16.90:1, 1
6.95:1, 17.00:1, 17.05:1, 17.10:1, 17.15:1, 17.20:1, 17.25:1, 17.30:1, 17.35:1,
17.40:1, 17.45:1, 17.50:1, 17.55:1, 17.60:1, 17.65:1, 17.70:1, 17.75:1, 17.80:1,
17.85:1, 17.90:1, 17.95:1, 18.00:1, 18.05:1, 18.10:1, 18.15:1, 18.20:1, 18.25:1
, 18.30:1, 18.35:1, 18.40:1, 18.45:1, 18.50:1, 18.55:1, 18.60:1, 18.65:1, 18.70:
1, 18.75:1, 18.80:1, 18.85:1, 18.90:1, 18.95:1, 19.00:1, 19.05:1, 19.10:1, 19.15
:1, 19.20:1, 19.25:1, 19.30:1, 19.35:1, 19.40:1, 19.45:1, 19.50:1, 19.55:1, 19.6
0:1, 19.65:1, 19.70:1, 19.75:1, 19.80:1, 19.85:1, 19.90:1, 19.95:1, 20.00:1, 20.
05:1, 20.10:1, 20.15:1, 20.20:1, 20.25:1, 20.30:1, 20.35:1, 20.40:1, 20.45:1, 20
.50:1, 20.55:1, 20.60:1, 20.65:1, 20.70:1, 20.75:1, 20.80:1, 20.85:1, 20.90:1, 2
0.95:1, 21.00:1, 21.05:1, 21.10:1, 21.15:1, 21.20:1, 21.25:1, 21.30:1, 21.35:1,
21.40:1, 21.45:1, 21.50:1, 21.55:1, 21.60:1, 21.65:1, 21.70:1, 21.75:1, 21.80:1,
21.85:1, 21.90:1, 21.95:1, 22.00:1, 22.05:1, 22.10:1, 22.15:1, 22.20:1, 22.25:1
, 22.30:1, 22.35:1, 22.40:1, 22.45:1, 22.50:1, 22.55:1, 22.60:1, 22.65:1, 22.70:
1, 22.75:1, 22.80:1, 22.85:1, 22.90:1, 22.95:1, 23.00, 23.05:1, 23.10:1, 23.15:1
, 23.20:1, 23.25:1, 23.30:1, 23.35:1, 23.40:1, 23.45:1, 23.50:1, 23.55:1, 23.60:
1, 23.65:1, 23.70:1, 23.75:1, 23.80:1, 23.85:1, 23.90:1, 23.95:1, 24.00:1, 24.05
:1, 24.10:1, 24.15:1, 24.20:1, 24.25:1, 24.30:1, 24.35:1, 24.40:1, 24.45:1, 24.5
0:1, 24.55:1, 24.60:1, 24.65:1, 24.70:1, 24.75:1, 24.80:1, 24.85:1, 24.90:1, 24.
95:1, 25.05:1, 25.10:1, 25.15:1, 25.20:1, 25.25:1, 25.30:1, 25.35:1, 25.40:1, 25
.45:1, 25.50:1, 25.55:1, 25.60:1, 25.65:1, 25.70:1, 25.75:1, 25.80:1, 25.85:1, 2
5.90:1, 25.95:1, 26.00:1, 26.05:1, 26.10:1, 26.15:1, 26.20:1, 26.25:1, 26.30:1,
26.35:1, 26.40:1, 26.45:1, 26.50:1, 26.55:1, 26.60:1, 26.65:1, 26.70:1, 26.75:1,
26.80:1, 26.85:1, 26.90:1, 26.95:1, 27.00:1, 27.05:1, 27.10:1, 27.15:1, 27.20:1
, 27.25:1, 27.30:1, 27.35:1, 27.40:1, 27.45:1, 27.50:1, 27.55:1, 27.60:1, 27.65:
1, 27.70:1, 27.75:1, 27.80:1, 27.85:1, 27.90:1, 27.95:1, 28.00:1, 28.05:1, 28.10
:1, 28.15:1, 28.20:1, 28.25:1, 28.30:1, 28.35:1, 28.40:1, 28.45:1, 28.50:1, 28.5
5:1, 28.60:1, 28.65:1, 28.70:1, 28.75:1, 28.80:1, 28.85:1, 28.90:1, 28.95:1, 29.
00:1, 29.05:1, 29.10:1, 29.15:1, 29.20:1, 29.25:1, 29.30:1, 29.35:1, 29.40:1, 29
.45:1, 29.50:1, 29.55:1, 29.60:1, 29.65:1, 29.70:1, 29.75:1, 29.80:1, 29.85:1, 2
9.90:1, 29.95:1, 30.00:1, 30.05:1, 30.10:1, 30.15:1, 30.20:1, 30.25:1, 30.30:1,
30.35:1, 30.40:1, 30.45:1, 30.50:1, 30.55:1, 30.60:1, 30.65:1, 30.70:1, 30.75:1,
30.80:1, 30.85:1, 30.90:1, 30.95:1, 31.00:1, 31.05:1, 31.10:1, 31.15:1, 31.20:1
, 31.25:1, 31.30:1, 31.35:1, 31.40:1, 31.45:1, 31.50:1, 31.55:1, 31.60:1, 31.65:
1, 31.70:1, 31.75:1, 31.80:1, 31.85:1, 31.90:1, 31.95:1, 32.00:1, 32.05:1, 32.10
:1, 32.15:1, 32.20:1, 32.25:1, 32.30:1, 32.35:1, 32.40:1, 32.45:1, 32.50:1, 32.5
5:1, 32.60:1, 32.65:1, 32.70:1, 32.75:1, 32.80:1, 32.85:1, 32.90:1, 32.95:1, 33.
00:1, 33.05:1, 33.10:1, 33.15:1, 33.20:1, 33.25:1, 33.30:1, 33.35:1, 33.40:1, 33
.45:1, 33.50:1, 33.55:1, 33.60:1, 33.65:1, 33.70:1, 33.75:1, 33.80:1, 33.85:1, 3
3.90:1, 33.95:1, 34.00:1, 34.05:1, 34.10:1, 34.15:1, 34.20:1, 34.25:1, 34.30:1,
34.35:1, 34.40:1, 34.45:1, 34.50:1, 34.55:1, 34.60:1, 34.65:1, 34.70:1, 34.75:1,
34.80:1, 34.85:1, 34.90:1, 34.95:1, 35.00:1, 35.05:1, 35.10:1, 35.15:1, 35.20:1
, 35.25:1, 35.30:1, 35.35:1, 35.40:1, 35.45:1, 35.50:1, 35.55:1, 35.60:1, 35.65:
1, 35.70:1, 35.75:1, 35.80:1, 35.85:1, 35.90:1, 35.95:1, 36.00:1, 36.05:1, 36.10
:1, 36.15:1, 36.20:1, 36.25:1, 36.30:1, 36.35:1, 36.40:1, 36.45:1, 36.50:1, 36.5
CD34 ⁺ CD90 to lymphocytes in a ratio of 5:1, 36.60:1, 36.65:1, 36.70:1, 36.75:1, 36.80:1, 36.85:1, 36.90:1, 36.95:1, or 37.00 ⁺ May be enriched with CD45RA ^- cells. In some embodiments, the peripheral blood of the donor is in a ratio of about 8.0:1 to about 10.0:1, e.g., about 8.00:1, 8.05:1, 8.10:1, 8.15:1, 8.20:1, 8.25:1, 8.30:1, 8.35:1, 8.40:1,
8.45:1, 8.50:1, 8.55:1, 8.60:1, 8.65:1, 8.70:1, 8.75:1, 8.80:1, 8.85:1, 8.90:1,
8.95:1, 9.00:1, 9.05:1, 9.10:1, 9.15:1, 9.20:1, 9.25:1, 9.30:1, 9.35:1, 9.40:1,
9.45:1, 9.50:1, 9.55:1, 9.60:1, 9.65:1, 9.70:1, 9.75:1, 9.80:1, 9.85:1, 9.90:1,
Enrich with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to lymphocytes to a ratio of 9.95:1, or 10.00:1. In some embodiments, the donor's peripheral blood is enriched with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to lymphocytes at a ratio of about 9.3:1.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising at least about
Density of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells of 16,000 cells/ml, e.g., about 20,000 cells/ml to about 75,000
cells/ml, about 25,000 cells/ml to about 70,000 cells/ml, about 30,000 cells/ml to about 65,000 cells/ml, about 3
5,000 cells/ml to about 60,000 cells/ml, about 40,000 cells/ml to about 55,000 cells/ml, or about 45,000 cells/ml to about 50,000 cells/ml (e.g., about 16,000 cells/ml, 17,000 cells/ml, 18,000 cells/ml cells/ml, 1
9,000 cells/ml, 20,000 cells/ml, 21,000 cells/ml, 22,000 cells/ml, 23,000 cells/ml, 24,000
cells/ml, 25,000 cells/ml, 26,000 cells/ml, 27,000 cells/ml, 28,000 cells/ml, 29,000 cells/
ml, 30,000 cells/ml, 31,000 cells/ml, 32,000 cells/ml, 33,000 cells/ml, 34,000 cells/ml, 3
5,000 cells/ml, 36,000 cells/ml, 37,000 cells/ml, 38,000 cells/ml, 39,000 cells/ml, 40,000
cells/ml, 41,000 cells/ml, 42,000 cells/ml, 43,000 cells/ml, 44,000 cells/ml, 45,000 cells/
ml, 46,000 cells/ml, 47,000 cells/ml, 48,000 cells/ml, 49,000 cells/ml, 50,000 cells/ml, 5
1,000 cells/ml, 52,000 cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000
cells/ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000 cells/ml, 61,000 cells/
ml, 62,000 cells/ml, 63,000 cells/ml, 64,000 cells/ml, 65,000 cells/ml, 66,000 cells/ml, 6
7,000 cells/ml, 68,000 cells/ml, 69,000 cells/ml, 70,000 cells/ml, 71,000 cells/ml, 72,000
cells/ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000 cells/ml, 77,000 cells/
ml, or more) with a density of CD34 ⁺ CD90 ⁺ CD45RA ^- cells, and approximately 2.4 x 10 ⁷ cells/m
Lymphocyte density of less than l, e.g. about 1 x 10 ⁷ cells/ml to about 2.3 x 10 ⁷ cells/ml, about 1.3 x 10 ⁷
cells/ml to about 2.1 x 10 ⁷ cells/ml, or about 1.5 x 10 ⁷ cells/ml to about 1.9 x 10 ⁷ cells/ml (e.g., about 2.4 x 10 ⁷ cells/ml, 2.3 x 10 ⁷ cells/ml) , 2.2 x 0 ⁷ cells/ml, 2.1 x 10 ⁷ cells/ml, 2 x
10 ⁷ cells/ml, 1.9 x 10 ⁷ cells/ml, 1.8 x 10 ⁷ cells/ml, 1.7 x 10 ⁷ cells/ml, 1.6 x 10 ⁷ cells
/ml, 1.5 x 10 ⁷ cells/ml 1.4 x 10 ⁷ cells/ml, 1.3 x 10 ⁷ cells/ml, 1.2 x 10 ⁷ cells/ml, 1.1
x 10 ⁷ cells/ml, 1 x 10 ⁷ cells/ml, or less, 0.9 x 10 ⁷ cells/ml, 0.8 x 10 ⁷ cells/ml, less) in sufficient quantities to produce
administering to said donor a CXCR2 agonist and a CXCR4 antagonist. In some embodiments, the method provides between about 20,000 cells/ml and about 75,000 cells/ml of CD34 ⁺ CD90 ⁺ CD45RA ^-
the CXCR2 agonist and CXCR4 antagonist in an amount sufficient to produce a cell population having a density of lymphocytes and a density of lymphocytes of about 1 x 10 ⁷ cells/ml to about 2.3 x 10 ⁷ cells/ml. including administering to the donor. In some embodiments, the method provides a method for producing cells from about 30,000 cells/ml to about 30,000 cells/ml.
with a density of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells of approximately 60,000 cells/ml, and from approximately 1.3 × 10 ⁷ cells/ml
CXCR2 in sufficient amounts to produce a cell population with a density of lymphocytes of approximately 2.3 x 10 ⁷ cells/ml.
agonist and CXCR4 antagonist to said donor. In some embodiments, the method has a density of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells of about 40,000 cells/ml to about 50,000 cells/ml, and about 1.5×10 ⁷ cells/ml to about 2×10 ⁷ administering to said donor a CXCR2 agonist and a CXCR4 antagonist in an amount sufficient to produce a cell population having a density of lymphocytes of 500 cells/ml.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising a CXCR2 agonist and a CXCR4 antagonist. , sufficient to produce a population of cells having a ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to monocytes in a sample of peripheral blood of the donor from about 0.0028 to about 0.0130 after administration of the CXCR2 agonist and the CXCR4 antagonist. and administering to said donor in an amount that is suitable for said donor. In some embodiments, CD34 ⁺ C on monocytes in said sample
The ratio of D90 ⁺ CD45RA ⁻ cells is approximately 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.0028
5, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.002
94, 0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.00301, 0.00302, 0.00
303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311, 0.0
0312, 0.00313, 0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.
00321, 0.00322, 0.00323, 0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00329, 0
.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.00337, 0.00338,
0.00339, 0.00340, 0.00341, 0.00342, 0.00343, 0.00344, 0.00345, 0.00346, 0.00347,
0.00348, 0.00349, 0.00350, 0.00351, 0.00352, 0.00353, 0.00354, 0.00355, 0.00356
, 0.00357, 0.00358, 0.00359, 0.00360, 0.00361, 0.00362, 0.00363, 0.00364, 0.0036
5, 0.00366, 0.00367, 0.00368, 0.00369, 0.00370, 0.00371, 0.00372, 0.00373, 0.003
74, 0.00375, 0.00376, 0.00378, 0.00379, 0.00380, 0.00381, 0.00382, 0.00383, 0.00
384, 0.00385, 0.00386, 0.00387, 0.00388, 0.00389, 0.00390, 0.00391, 0.00392, 0.0
0393, 0.00394, 0.00395, 0.00396, 0.00397, 0.00398, 0.00399, 0.00401, 0.00402, 0.
00403, 0.00404, 0.00405, 0.00406, 0.00407, 0.00408, 0.00409, 0.00410, 0.00411, 0
.00412, 0.00413, 0.00414, 0.00415, 0.00416, 0.00417, 0.00418, 0.00419, 0.00420,
0.00421, 0.00422, 0.00423, 0.00424, 0.00425, 0.00426, 0.00427, 0.00428, 0.00429,
0.00430, 0.00431, 0.00432, 0.00433, 0.00434, 0.00435, 0.00436, 0.00437, 0.00438
, 0.00439, 0.00440, 0.00441, 0.00442, 0.00443, 0.00444, 0.00445, 0.00446, 0.0044
7, 0.00448, 0.00449, 0.00450, 0.00451, 0.00452, 0.00453, 0.00454, 0.00455, 0.004
56, 0.00457, 0.00458, 0.00459, 0.00460, 0.00461, 0.00462, 0.00463, 0.00464, 0.00
465, 0.00466, 0.00467, 0.00468, 0.00469, 0.00470, 0.00471, 0.00472, 0.00473, 0.0
0474, 0.00475, 0.00476, 0.00478, 0.00479, 0.00480, 0.00481, 0.00482, 0.00483, 0.
00484, 0.00485, 0.00486, 0.00487, 0.00488, 0.00489, 0.00490, 0.00491, 0.00492, 0
.00493, 0.00494, 0.00495, 0.00496, 0.00497, 0.00498, 0.00499, 0.00500, 0.00501,
0.00502, 0.00503, 0.00504, 0.00505, 0.00506, 0.00507, 0.00508, 0.00509, 0.00510,
0.00511, 0.00512, 0.00513, 0.00514, 0.00515, 0.00516, 0.00517, 0.00518, 0.00519
, 0.00520, 0.00521, 0.00522, 0.00523, 0.00524, 0.00525, 0.00526, 0.00527, 0.0052
8, 0.00529, 0.00530, 0.00531, 0.00532, 0.00533, 0.00534, 0.00535, 0.00536, 0.005
37, 0.00538, 0.00539, 0.00540, 0.00541, 0.00542, 0.00543, 0.00544, 0.00545, 0.00
546, 0.00547, 0.00548, 0.00549, 0.00550, 0.00551, 0.00552, 0.00553, 0.00554, 0.0
0555, 0.00556, 0.00557, 0.00558, 0.00559, 0.00560, 0.00561, 0.00562, 0.00563, 0.
00564, 0.00565, 0.00566, 0.00567, 0.00568, 0.00569, 0.00570, 0.00571, 0.00572, 0
.00573, 0.00574, 0.00575, 0.00576, 0.00578, 0.00579, 0.00580, 0.00581, 0.00582,
0.00583, 0.00584, 0.00585, 0.00586, 0.00587, 0.00588, 0.00589, 0.00590, 0.00591,
0.00592, 0.00593, 0.00594, 0.00595, 0.00596, 0.00597, 0.00598, 0.00599, 0.00600
, 0.00601, 0.00602, 0.00603, 0.00604, 0.00605, 0.00606, 0.00607, 0.00608, 0.0060
9, 0.00610, 0.00611, 0.00612, 0.00613, 0.00614, 0.00615, 0.00616, 0.00617, 0.006
18, 0.00619, 0.00620, 0.00621, 0.00622, 0.00623, 0.00624, 0.00625, 0.00626, 0.00
627, 0.00628, 0.00629, 0.00630, 0.00631, 0.00632, 0.00633, 0.00634, 0.00635, 0.0
0636, 0.00637, 0.00638, 0.00639, 0.00640, 0.00641, 0.00642, 0.00643, 0.00644, 0.
00645, 0.00646, 0.00647, 0.00648, 0.00649, 0.00650, 0.00651, 0.00652, 0.00653, 0
.00654, 0.00655, 0.00656, 0.00657, 0.00658, 0.00659, 0.00660, 0.00661, 0.00662,
0.00663, 0.00664, 0.00665, 0.00666, 0.00667, 0.00668, 0.00669, 0.00670, 0.00671,
0.00672, 0.00673, 0.00674, 0.00675, 0.00676, 0.00678, 0.00679, 0.00680, 0.00681
, 0.00682, 0.00683, 0.00684, 0.00685, 0.00686, 0.00687, 0.00688, 0.00689, 0.0069
0, 0.00691, 0.00692, 0.00693, 0.00694, 0.00695, 0.00696, 0.00697, 0.00698, 0.006
99, 0.00700, 0.00701, 0.00702, 0.00703, 0.00704, 0.00705, 0.00706, 0.00707, 0.00
708, 0.00709, 0.00710, 0.00711, 0.00712, 0.00713, 0.00714, 0.00715, 0.00716, 0.0
0717, 0.00718, 0.00719, 0.00720, 0.00721, 0.00722, 0.00723, 0.00724, 0.00725, 0.
00726, 0.00727, 0.00728, 0.00729, 0.00730, 0.00731, 0.00732, 0.00733, 0.00734, 0
.00735, 0.00736, 0.00737, 0.00738, 0.00739, 0.00740, 0.00741, 0.00742, 0.00743,
0.00744, 0.00745, 0.00746, 0.00747, 0.00748, 0.00749, 0.00750, 0.00751, 0.00752,
0.00753, 0.00754, 0.00755, 0.00756, 0.00757, 0.00758, 0.00759, 0.00760, 0.00761
, 0.00762, 0.00763, 0.00764, 0.00765, 0.00766, 0.00767, 0.00768, 0.00769, 0.0077
0, 0.00771, 0.00772, 0.00773, 0.00774, 0.00775, 0.00776, 0.00777, 0.00778, 0.007
79, 0.00780, 0.00781, 0.00782, 0.00783, 0.00784, 0.00785, 0.00786, 0.00787, 0.00
788, 0.00789, 0.00790, 0.00791, 0.00792, 0.00793, 0.00794, 0.00795, 0.00796, 0.0
0797, 0.00798, 0.00799, 0.00800, 0.00801, 0.00802, 0.00803, 0.00804, 0.00805, 0.
00806, 0.00807, 0.00808, 0.00809, 0.00810, 0.00811, 0.00812, 0.00813, 0.00814, 0
.00815, 0.00816, 0.00817, 0.00818, 0.00819, 0.00820, 0.00821, 0.00822, 0.00823,
0.00824, 0.00825, 0.00826, 0.00827, 0.00828, 0.00829, 0.00830, 0.00831, 0.00832,
0.00833, 0.00834, 0.00835, 0.00836, 0.00837, 0.00838, 0.00839, 0.00840, 0.00841
, 0.00842, 0.00843, 0.00844, 0.00845, 0.00846, 0.00847, 0.00848, 0.00849, 0.0085
0, 0.00851, 0.00852, 0.00853, 0.00854, 0.00855, 0.00856, 0.00857, 0.00858, 0.008
59, 0.00860, 0.00861, 0.00862, 0.00863, 0.00864, 0.00865, 0.00866, 0.00867, 0.00
868, 0.00869, 0.00870, 0.00871, 0.00872, 0.00873, 0.00874, 0.00875, 0.00876, 0.0
0877, 0.00878, 0.00879, 0.00880, 0.00881, 0.00882, 0.00883, 0.00884, 0.00885, 0.
00886, 0.00887, 0.00888, 0.00889, 0.00890, 0.00891, 0.00892, 0.00893, 0.00894, 0
.00895, 0.00896, 0.00897, 0.00898, 0.00899, 0.00900, 0.00901, 0.00902, 0.00903,
0.00904, 0.00905, 0.00906, 0.00907, 0.00908, 0.00909, 0.00910, 0.00911, 0.00912,
0.00913, 0.00914, 0.00915, 0.00916, 0.00917, 0.00918, 0.00919, 0.00920, 0.00921
, 0.00922, 0.00923, 0.00924, 0.00925, 0.00926, 0.00927, 0.00928, 0.00929, 0.0093
0, 0.00931, 0.00932, 0.00933, 0.00934, 0.00935, 0.00936, 0.00937, 0.00938, 0.009
39, 0.00940, 0.00941, 0.00942, 0.00943, 0.00944, 0.00945, 0.00946, 0.00947, 0.00
948, 0.00949, 0.00950, 0.00951, 0.00952, 0.00953, 0.00954, 0.00955, 0.00956, 0.0
0957, 0.00958, 0.00959, 0.00960, 0.00961, 0.00962, 0.00963, 0.00964, 0.00965, 0.
00966, 0.00967, 0.00968, 0.00969, 0.00970, 0.00971, 0.00972, 0.00973, 0.00974, 0
.00975, 0.00976, 0.00977, 0.00978, 0.00979, 0.00980, 0.00981, 0.00982, 0.00983,
0.00984, 0.00985, 0.00986, 0.00987, 0.00988, 0.00989, 0.00990, 0.00991, 0.00992,
0.00993, 0.00994, 0.00995, 0.00996, 0.00997, 0.00998, 0.00999, 0.0100, 0.0101,
0.0103, 0.0104, 0.0105, 0.0106, 0.0107, 0.0108, 0.0109, 0.0110, 0.0111, 0.0112,
0.0113, 0.0114, 0.0115, 0.0116, 0.0117, 0.0118, 0.0119, 0.0120, 0.0121, 0.0122,
May be 0.0123, 0.0124, 0.0125, 0.0126, 0.0127, 0.0128, 0.0129, or 0.0130. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to monocytes in the sample is about 0.0063 to about 0.0083, e.g., about 0.00630, 0.00631, 0.00632, 0.00633, 0.
00634, 0.00635, 0.00636, 0.00637, 0.00638, 0.00639, 0.00640, 0.00641, 0.00642, 0
.00643, 0.00644, 0.00645, 0.00646, 0.00647, 0.00648, 0.00649, 0.00650, 0.00651,
0.00652, 0.00653, 0.00654, 0.00655, 0.00656, 0.00657, 0.00658, 0.00659, 0.00660,
0.00661, 0.00662, 0.00663, 0.00664, 0.00665, 0.00666, 0.00667, 0.00668, 0.00669
, 0.00670, 0.00671, 0.00672, 0.00673, 0.00674, 0.00675, 0.00676, 0.00678, 0.0067
9, 0.00680, 0.00681, 0.00682, 0.00683, 0.00684, 0.00685, 0.00686, 0.00687, 0.006
88, 0.00689, 0.00690, 0.00691, 0.00692, 0.00693, 0.00694, 0.00695, 0.00696, 0.00
697, 0.00698, 0.00699, 0.00700, 0.00701, 0.00702, 0.00703, 0.00704, 0.00705, 0.0
0706, 0.00707, 0.00708, 0.00709, 0.00710, 0.00711, 0.00712, 0.00713, 0.00714, 0.
00715, 0.00716, 0.00717, 0.00718, 0.00719, 0.00720, 0.00721, 0.00722, 0.00723, 0
.00724, 0.00725, 0.00726, 0.00727, 0.00728, 0.00729, 0.00730, 0.00731, 0.00732,
0.00733, 0.00734, 0.00735, 0.00736, 0.00737, 0.00738, 0.00739, 0.00740, 0.00741,
0.00742, 0.00743, 0.00744, 0.00745, 0.00746, 0.00747, 0.00748, 0.00749, 0.00750
, 0.00751, 0.00752, 0.00753, 0.00754, 0.00755, 0.00756, 0.00757, 0.00758, 0.0075
9, 0.00760, 0.00761, 0.00762, 0.00763, 0.00764, 0.00765, 0.00766, 0.00767, 0.007
68, 0.00769, 0.00770, 0.00771, 0.00772, 0.00773, 0.00774, 0.00775, 0.00776, 0.00
777, 0.00778, 0.00779, 0.00780, 0.00781, 0.00782, 0.00783, 0.00784, 0.00785, 0.0
0786, 0.00787, 0.00788, 0.00789, 0.00790, 0.00791, 0.00792, 0.00793, 0.00794, 0.
00795, 0.00796, 0.00797, 0.00798, 0.00799, 0.00800, 0.00801, 0.00802, 0.00803, 0
.00804, 0.00805, 0.00806, 0.00807, 0.00808, 0.00809, 0.00810, 0.00811, 0.00812,
0.00813, 0.00814, 0.00815, 0.00816, 0.00817, 0.00818, 0.00819, 0.00820, 0.00821,
0.00822, 0.00823, 0.00824, 0.00825, 0.00826, 0.00827, 0.00828, 0.00829, 0.00830
, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to monocytes in the sample. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to monocytes in said sample is about 0.
It is .0073.

In a further aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising
Agonists and CXCR4 antagonists are evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and the CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and the CXCR4 antagonist. said donor in an amount sufficient to enrich said donor's peripheral blood with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to monocytes at a ratio of about 1.5:1 to about 8.5:1. including administering. In some embodiments, the peripheral blood of the donor is about 1.50:1, 1.55:1, 1.60:1, 1.65:1, 1.70:1,
1.75:1, 1.80:1, 1.85:1, 1.90:1, 1.95:1, 2.00:1, 2.05:1, 2.10:1, 2.15:1, 2.20:1,
2.25:1, 2.30:1, 2.35:1, 2.40:1, 2.45:1, 2.50:1, 2.55:1, 2.60:1, 2.65:1, 2.70:1,
2.75:1, 2.80:1, 2.85:1, 2.90:1, 2.95:1, 3.00:1, 3.05:1, 3.10:1, 3.15:1, 3.20:1,
3.25:1, 3.30:1, 3.35:1, 3.40:1, 3.45:1, 3.50:1, 3.55:1, 3.60:1, 3.65:1, 3.70:1,
3.75:1, 3.80:1, 3.85:1, 3.90:1, 3.95:1, 4.00:1, 4.05:1, 4.10:1, 4.15:1, 4.20:1,
4.25:1, 4.30:1, 4.35:1, 4.40:1, 4.45:1, 4.50:1, 4.55:1, 4.60:1, 4.65:1, 4.70:1,
4.75:1, 4.80:1, 4.85:1, 4.90:1, 4.95:1, 5.00:1, 5.05:1, 5.10:1, 5.15:1, 5.20:1,
5.25:1, 5.30:1, 5.35:1, 5.40:1, 5.45:1, 5.50:1, 5.55:1, 5.60:1, 5.65:1, 5.70:1,
5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1, 6.15:1, 6.20:1,
6.25:1, 6.30:1, 6.35:1, 6.40:1, 6.45:1, 6.50:1, 6.55:1, 6.60:1, 6.65:1, 6.70:1,
6.75:1, 6.80:1, 6.85:1, 6.90:1, 6.95:1, 7.00:1, 7.05:1, 7.10:1, 7.15:1, 7.20:1,
7.25:1, 7.30:1, 7.35:1, 7.40:1, 7.45:1, 7.50:1, 7.55:1, 7.60:1, 7.65:1, 7.70:1,
7.75:1, 7.80:1, 7.85:1, 7.90:1, 7.95:1, 8.00:1, 8.05:1, 8.10:1, 8.15:1, 8.20:1,
CD34 ⁺ CD to monocytes in a ratio of 8.25:1, 8.30:1, 8.35:1, 8.40:1, 8.45:1, or 8.50:1
The donor's peripheral blood may be enriched with 90 ⁺ CD45RA ⁻ cells. In some embodiments, the donor's peripheral blood is enriched with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to monocytes to a ratio of about 1.9:1.

In another aspect, the invention provides methods for obtaining peripheral blood from the bone marrow of a mammalian donor (e.g., a human donor).
The present invention relates to a method for mobilizing a population of hematopoietic stem cells into a blood cell, the method comprising at least about
16,000 cells/ml CD34⁺CD90⁺ CD45RA^-Cell density, e.g. about 20,000 cells/ml to about 75,000
cells/ml, about 25,000 cells/ml to about 70,000 cells/ml, about 30,000 cells/ml to about 65,000 cells/ml, about 3
5,000 cells/ml to approximately 60,000 cells/ml, approximately 40,000 cells/ml to approximately 55,000 cells/ml, or approximately 45,000 cells/ml
cells/ml to approximately 50,000 cells/ml (e.g. approximately 16,000 cells/ml, 17,000 cells/ml, 18,000 cells/ml, 1
9,000 cells/ml, 20,000 cells/ml, 21,000 cells/ml, 22,000 cells/ml, 23,000 cells/ml, 24,000
cells/ml, 25,000 cells/ml, 26,000 cells/ml, 27,000 cells/ml, 28,000 cells/ml, 29,000 cells/
ml, 30,000 cells/ml, 31,000 cells/ml, 32,000 cells/ml, 33,000 cells/ml, 34,000 cells/ml, 3
5,000 cells/ml, 36,000 cells/ml, 37,000 cells/ml, 38,000 cells/ml, 39,000 cells/ml, 40,000
cells/ml, 41,000 cells/ml, 42,000 cells/ml, 43,000 cells/ml, 44,000 cells/ml, 45,000 cells/
ml, 46,000 cells/ml, 47,000 cells/ml, 48,000 cells/ml, 49,000 cells/ml, 50,000 cells/ml, 5
1,000 cells/ml, 52,000 cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000
cells/ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000 cells/ml, 61,000 cells/
ml, 62,000 cells/ml, 63,000 cells/ml, 64,000 cells/ml, 65,000 cells/ml, 66,000 cells/ml, 6
7,000 cells/ml, 68,000 cells/ml, 69,000 cells/ml, 70,000 cells/ml, 71,000 cells/ml, 72,000
cells/ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000 cells/ml, 77,000 cells/
ml, or more) of CD34⁺ CD90⁺ CD45RA^-with a density of cells, and approximately 6 x 10⁶cells/ml
Density of monocytes below, e.g. 3.4 x 10⁶cells/ml ~ approx. 5.9 x 10⁶cells/ml, approximately 3.5 x 10⁶cell/
ml ~ approx. 5.7 x 10⁶cells/ml, or about 4 x 10⁶cells/ml ~ approx. 5 x 10⁶cells/ml (e.g., 5.9 x 10⁶
cells/ml, 5.8 x 10⁶cells/ml, 5.7 x 10⁶cells/ml, 5.6 x 10⁶cells/ml, 5.5 x 10⁶cells/ml,
5.4 x 10⁶cells/ml, 5.3 x 10⁶cells/ml, 5.2 x 10⁶cells/ml, 5.1 x 10⁶cells/ml, 5 x 10⁶
cells/ml, 4.9 x 10⁶cells/ml, 4.8 x 10⁶cells/ml, 4.7 x 10⁶cells/ml, 4.6 x 10⁶cells/ml,
4.5 x 10⁶cells/ml, 4.4 x 10⁶cells/ml, 4.3 x 10⁶cells/ml, 4.2 x 10⁶cells/ml, 4.1 x 1
0⁶cells/ml, 4 x 10⁶cells/ml, 3.9 x 10⁶cells/ml, 3.8 x 10⁶cells/ml, 3.7 x 10⁶cells/ml,
3.6x10⁶cells/ml, 3.5 x 10⁶cells/ml, 3.4 x 10⁶cells/ml, or less) of monocytes
CXCR2 agonists and CXCR4 antagonists in amounts sufficient to produce a dense population of cells.
agonist to said donor. In some embodiments, the method comprises about 20
,000 cells/ml to approximately 75,000 cells/ml of CD34⁺CD90⁺ CD45RA^-with cell density, and approximately 3.4x
Ten⁶cells/ml ~ approx. 6×10⁶in amounts sufficient to produce a cell population with a density of monocytes cells/ml.
, administering to said donor a CXCR2 agonist and a CXCR4 antagonist. how many
In such embodiments, the method provides between about 30,000 cells/ml and about 60,000 cells/ml of CD34 cells/ml.⁺CD90⁺ CD45RA^-
with a density of cells, and approximately 4 x 10⁶cells/ml ~ approx. 5.5×10⁶with a density of monocytes in cells/ml
CXCR2 agonist and CXCR4 antagonist in sufficient amounts to produce a population of cells.
This includes administering to the patient. In some embodiments, the method provides between about 40,000 cells/ml and about 5
0,000 cells/ml CD34⁺CD90⁺ CD45RA^-with a density of cells, and approximately 4 x 10⁶cells/ml ~ approx. 5×1
0⁶CXCR2 agonist and
and a CXCR4 antagonist to said donor.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising a CXCR2 agonist and a CXCR4 antagonist. , CD34 ⁺ CD90 ⁺ CD45RA on CD34 ⁺ cells in a sample of peripheral blood of said donor after administration of said CXCR2 agonist and said CXCR4 antagonist.
^- administering to said donor an amount sufficient to produce a population of cells having a cell ratio of about 0.393 to about 0.745. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to CD34 ⁺ cells in the sample is about 0.393, 0.394, 0.395, 0.396, 0.397, 0.398, 0
.399, 0.401, 0.402, 0.403, 0.404, 0.405, 0.406, 0.407, 0.408, 0.409, 0.410, 0.41
1, 0.412, 0.413, 0.414, 0.415, 0.416, 0.417, 0.418, 0.419, 0.420, 0.421, 0.422,
0.423, 0.424, 0.425, 0.426, 0.427, 0.428, 0.429, 0.430, 0.431, 0.432, 0.433, 0.4
34, 0.435, 0.436, 0.437, 0.438, 0.439, 0.440, 0.441, 0.442, 0.443, 0.444, 0.445,
0.446, 0.447, 0.448, 0.449, 0.450, 0.451, 0.452, 0.453, 0.454, 0.455, 0.456, 0.
457, 0.458, 0.459, 0.460, 0.461, 0.462, 0.463, 0.464, 0.465, 0.466, 0.467, 0.468
, 0.469, 0.470, 0.471, 0.472, 0.473, 0.474, 0.475, 0.476, 0.478, 0.479, 0.480, 0
.481, 0.482, 0.483, 0.484, 0.485, 0.486, 0.487, 0.488, 0.489, 0.490, 0.491, 0.49
2, 0.493, 0.494, 0.495, 0.496, 0.497, 0.498, 0.499, 0.500, 0.501, 0.502, 0.503,
0.504, 0.505, 0.506, 0.507, 0.508, 0.509, 0.510, 0.511, 0.512, 0.513, 0.514, 0.5
15, 0.516, 0.517, 0.518, 0.519, 0.520, 0.521, 0.522, 0.523, 0.524, 0.525, 0.526,
0.527, 0.528, 0.529, 0.530, 0.531, 0.532, 0.533, 0.534, 0.535, 0.536, 0.537, 0.
538, 0.539, 0.540, 0.541, 0.542, 0.543, 0.544, 0.545, 0.546, 0.547, 0.548, 0.549
, 0.550, 0.551, 0.552, 0.553, 0.554, 0.555, 0.556, 0.557, 0.558, 0.559, 0.560, 0
.561, 0.562, 0.563, 0.564, 0.565, 0.566, 0.567, 0.568, 0.569, 0.570, 0.571, 0.57
2, 0.573, 0.574, 0.575, 0.576, 0.578, 0.579, 0.580, 0.581, 0.582, 0.583, 0.584,
0.585, 0.586, 0.587, 0.588, 0.589, 0.590, 0.591, 0.592, 0.593, 0.594, 0.595, 0.5
96, 0.597, 0.598, 0.599, 0.600, 0.601, 0.602, 0.603, 0.604, 0.605, 0.606, 0.607,
0.608, 0.609, 0.610, 0.611, 0.612, 0.613, 0.614, 0.615, 0.616, 0.617, 0.618, 0.
619, 0.620, 0.621, 0.622, 0.623, 0.624, 0.625, 0.626, 0.627, 0.628, 0.629, 0.630
, 0.631, 0.632, 0.633, 0.634, 0.635, 0.636, 0.637, 0.638, 0.639, 0.640, 0.641, 0
.642, 0.643, 0.644, 0.645, 0.646, 0.647, 0.648, 0.649, 0.650, 0.651, 0.652, 0.65
3, 0.654, 0.655, 0.656, 0.657, 0.658, 0.659, 0.660, 0.661, 0.662, 0.663, 0.664,
0.665, 0.666, 0.667, 0.668, 0.669, 0.670, 0.671, 0.672, 0.673, 0.674, 0.675, 0.6
76, 0.678, 0.679, 0.680, 0.681, 0.682, 0.683, 0.684, 0.685, 0.686, 0.687, 0.688,
0.689, 0.690, 0.691, 0.692, 0.693, 0.694, 0.695, 0.696, 0.697, 0.698, 0.699, 0.
700, 0.701, 0.702, 0.703, 0.704, 0.705, 0.706, 0.707, 0.708, 0.709, 0.710, 0.711
, 0.712, 0.713, 0.714, 0.715, 0.716, 0.717, 0.718, 0.719, 0.720, 0.721, 0.722, 0
.723, 0.724, 0.725, 0.726, 0.727, 0.728, 0.729, 0.730, 0.731, 0.732, 0.733, 0.73
4, 0.735, 0.736, 0.737, 0.738, 0.739, 0.740, 0.741, 0.742, 0.743, 0.744, or 0.
It may be 745. In some embodiments, the CD for CD34 ⁺ cells in said sample
The ratio of 34 ⁺ CD90 ⁺ CD45RA ⁻ cells is about 0.625 to about 0.725, e.g., about 0.625, 0.626, 0.627,
0.628, 0.629, 0.630, 0.631, 0.632, 0.633, 0.634, 0.635, 0.636, 0.637, 0.638, 0.6
39, 0.640, 0.641, 0.642, 0.643, 0.644, 0.645, 0.646, 0.647, 0.648, 0.649, 0.650,
0.651, 0.652, 0.653, 0.654, 0.655, 0.656, 0.657, 0.658, 0.659, 0.660, 0.661, 0.
662, 0.663, 0.664, 0.665, 0.666, 0.667, 0.668, 0.669, 0.670, 0.671, 0.672, 0.673
, 0.674, 0.675, 0.676, 0.678, 0.679, 0.680, 0.681, 0.682, 0.683, 0.684, 0.685, 0
.686, 0.687, 0.688, 0.689, 0.690, 0.691, 0.692, 0.693, 0.694, 0.695, 0.696, 0.69
7, 0.698, 0.699, 0.700, 0.701, 0.702, 0.703, 0.704, 0.705, 0.706, 0.707, 0.708,
0.709, 0.710, 0.711, 0.712, 0.713, 0.714, 0.715, 0.716, 0.717, 0.718, 0.719, 0.7
C for CD34 ⁺ cells in said sample of 20, 0.721, 0.722, 0.723, 0.724, or 0.725
The ratio of D34 ⁺ CD90 ⁺ CD45RA ⁻ cells is, In some embodiments, the CD in said sample
The ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to 34 ⁺ cells is approximately 0.676.

In a further aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising
Agonists and CXCR4 antagonists are evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and the CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and the CXCR4 antagonist. said donor in an amount sufficient to enrich said donor's peripheral blood with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to CD34 ⁺ cells in a ratio of about 1.1:1 to about 4.8:1 when including administering to In some embodiments, the peripheral blood of the donor is about 1.10:1, 1.15:1, 1.20:1, 1.25:1, 1.30
:1, 1.35:1, 1.40:1, 1.45:1, 1.50:1, 1.55:1, 1.60:1, 1.65:1, 1.70:1, 1.75:1, 1.80
:1, 1.85:1, 1.90:1, 1.95:1, 2.00:1, 2.05:1, 2.10:1, 2.15:1, 2.20:1, 2.25:1, 2.30
:1, 2.35:1, 2.40:1, 2.45:1, 2.50:1, 2.55:1, 2.60:1, 2.65:1, 2.70:1, 2.75:1, 2.80
:1, 2.85:1, 2.90:1, 2.95:1, 3.00:1, 3.05:1, 3.10:1, 3.15:1, 3.20:1, 3.25:1, 3.30
:1, 3.35:1, 3.40:1, 3.45:1, 3.50:1, 3.55:1, 3.60:1, 3.65:1, 3.70:1, 3.75:1, 3.80
:1, 3.85:1, 3.90:1, 3.95:1, 4.00:1, 4.05:1, 4.10:1, 4.15:1, 4.20:1, 4.25:1, 4.30
:1, 4.35:1, 4.40:1, 4.45:1, 4.50:1, 4.55:1, 4.60:1, 4.65:1, 4.70:1, 4.75:1, or
The donor's peripheral blood may be enriched with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to CD34 ⁺ cells to a ratio of 4.80:1. In some embodiments, the donor's peripheral blood is enriched with CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to CD34 ⁺ cells to a ratio of about 1.2:1.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising a CXCR2 agonist and a CXCR4 antagonist. , after administration of said CXCR2 agonist and said CXCR4 antagonist, the frequency of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells in a sample of said donor's peripheral blood is approximately 0.
administering to said donor an amount sufficient to produce a population of cells that is .020% to about .110%. In some embodiments, the cell population has a frequency of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells of about 0.020%, 0.021%, 0.022%, 0.023%, 0.024%, 0.025%, 0.026%, 0.027%, 0.028%, 0.02
9%, 0.030%, 0.031%, 0.032%, 0.033%, 0.034%, 0.035%, 0.036%, 0.037%, 0.038%, 0.03
9%, 0.040%, 0.041%, 0.042%, 0.043%, 0.044%, 0.045%, 0.046%, 0.047%, 0.048%, 0.04
9%, 0.050%, 0.051%, 0.052%, 0.053%, 0.054%, 0.055%, 0.056%, 0.057%, 0.058%, 0.05
9%, 0.060%, 0.061%, 0.062%, 0.063%, 0.064%, 0.065%, 0.066%, 0.067%, 0.068%, 0.06
9%, 0.070%, 0.071%, 0.072%, 0.073%, 0.074%, 0.075%, 0.076%, 0.077%, 0.078%, 0.07
9%, 0.080%, 0.081%, 0.082%, 0.083%, 0.084%, 0.085%, 0.086%, 0.087%, 0.088%, 0.08
9%, 0.090%, 0.091%, 0.092%, 0.093%, 0.094%, 0.095%, 0.096%, 0.097%, 0.098%, 0.09
9%, 0.100%, 0.101%, 0.102%, 0.103%, 0.104%, 0.105%, 0.106%, 0.107%, 0.108%, 0.10
9%, or 0.110%. In some embodiments, the cell population is CD34 ⁺
The frequency of CD90 ⁺ CD45RA ⁻ cells is about 0.046% to about 0.086%, e.g., about 0.046%, 0.047%, 0.048%,
0.049%, 0.050%, 0.051%, 0.052%, 0.053%, 0.054%, 0.055%, 0.056%, 0.057%, 0.058%,
0.059%, 0.060%, 0.061%, 0.062%, 0.063%, 0.064%, 0.065%, 0.066%, 0.067%, 0.068%,
0.069%, 0.070%, 0.071%, 0.072%, 0.073%, 0.074%, 0.075%, 0.076%, 0.077%, 0.078%,
The frequency of hematopoietic stem cells is 0.079%, 0.080%, 0.081%, 0.082%, 0.083%, 0.084%, 0.085%, or 0.086%. In some embodiments, the population of cells is CD34 ⁺ CD90 ⁺ CD45RA ^-
The cell frequency is about 0.066%.

In a further aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising
administering an agonist and a CXCR4 antagonist to the donor, wherein a sample of peripheral blood of the donor after administration of the CXCR2 agonist and the CXCR4 antagonist is
The frequency of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells in peripheral blood of said donor is at least 3-fold (e.g., Approx. 5.1x to approx. 25.7x, e.g. approx. 5.1x, 5.2x, 5.3x
, 5.4x, 5.5x, 5.6x, 5.7x, 5.8x, 5.9x, 6.0x, 6.1x, 6.2x, 6.3x, 6.4x, 6
.5x, 6.6x, 6.7x, 6.8x, 6.9x, 7.0x, 7.1x, 7.2x, 7.3x, 7.4x, 7.5x, 7.6
times, 7.7 times, 7.8 times, 7.9 times, 8.0 times, 8.1 times, 8.2 times, 8.3 times, 8.4 times, 8.5 times, 8.6 times, 8.7 times,
8.8x, 8.9x, 9.0x, 9.1x, 9.2x, 9.3x, 9.4x, 9.5x, 9.6x, 9.7x, 9.8x, 9.
9x, 10.0x, 10.1x, 10.2x, 10.3x, 10.4x, 10.5x, 10.6x, 10.7x, 10.8x, 10.
9x, 11.0x, 11.1x, 11.2x, 11.3x, 11.4x, 11.5x, 11.6x, 11.7x, 11.8x, 11.
9x, 12.0x, 12.1x, 12.2x, 12.3x, 12.4x, 12.5x, 12.6x, 12.7x, 12.8x, 12.
9x, 13.0x, 13.1x, 13.2x, 13.3x, 13.4x, 13.5x, 13.6x, 13.7x, 13.8x, 13.
9x, 14.0x, 14.1x, 14.2x, 14.3x, 14.4x, 14.5x, 14.6x, 14.7x, 14.8x, 14.
9x, 15.0x, 15.1x, 15.2x, 15.3x, 15.4x, 15.5x, 15.6x, 15.7x, 15.8x, 15.
9x, 16.0x, 16.1x, 16.2x, 16.3x, 16.4x, 16.5x, 16.6x, 16.7x, 16.8x, 16.
9x, 17.0x, 17.1x, 17.2x, 17.3x, 17.4x, 17.5x, 17.6x, 17.7x, 17.8x, 17.
9x, 18.0x, 18.1x, 18.2x, 18.3x, 18.4x, 18.5x, 18.6x, 18.7x, 18.8x, 18.
9x, 19.0x, 19.1x, 19.2x, 19.3x, 19.4x, 19.5x, 19.6x, 19.7x, 19.8x, 19.
9x, 20.0x, 20.1x, 20.2x, 20.3x, 20.4x, 20.5x, 20.6x, 20.7x, 20.8x, 20.
9x, 21.0x, 21.1x, 21.2x, 21.3x, 21.4x, 21.5x, 21.6x, 21.7x, 21.8x, 21.
9x, 22.0x, 22.1x, 22.2x, 22.3x, 22.4x, 22.5x, 22.6x, 22.7x, 22.8x, 22.
9x, 23.0x, 23.1x, 23.2x, 23.3x, 23.4x, 23.5x, 23.6x, 23.7x, 23.8x, 23.
9x, 24.0x, 24.1x, 24.2x, 24.3x, 24.4x, 24.5x, 24.6x, 24.7x, 24.8x, 24.
administering the CXCR2 agonist and the CXCR4 antagonist to the donor in an amount sufficient to increase including doing. In some embodiments, after administration of said CXCR2 agonist and said CXCR4 antagonist, the frequency of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells in peripheral blood of said donor is about 5.1.
times to approximately 7.1 times, for example, approximately 5.1 times, 5.2 times, 5.3 times, 5.4 times, 5.5 times, 5.6 times, 5.7 times, 5.8 times, 5
.9x, 6.0x, 6.1x, 6.2x, 6.3x, 6.4x, 6.5x, 6.6x, 6.7x, 6.8x, 6.9x, 7.0
or 7.1 times. In some embodiments, the frequency of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells in peripheral blood of said donor increases about 5.8-fold after administration of a CXCR2 agonist and a CXCR4 antagonist.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into the peripheral blood, the method comprising administering to the donor a mobilizing amount of a CXCR2 agonist and a CXCR4 antagonist. administering; obtaining an input value for each of the one or more parameters in Table 2 characterizing a sample of peripheral blood of the donor; and determining that the input value for each of the one or more parameters is To expand hematopoietic stem cells ex vivo, if the corresponding reference standards for each of the above parameters are met,
or providing said sample for use in treating one or more stem cell disorders in a mammalian patient. In some embodiments, the one or more reference parameters are a combination of parameters listed in any one of Tables 3-6 herein.

In some embodiments of any of the above aspects of the invention, the sample is administered for about 3 hours to about 5 hours after administration of the CXCR2 agonist and CXCR4 antagonist (e.g., about 3 hours after administration of the CXCR2 agonist and CXCR4 antagonist). Time, 3.1 hours, 3.2 hours, 3.3 hours, 3.4 hours, 3.5
Time, 3.6 hours, 3.7 hours, 3.8 hours, 3.9 hours, 4.0 hours, 4.1 hours, 4.2 hours, 4.3 hours, 4.
4 hours, 4.5 hours, 4.6 hours, 4.7 hours, 4.8 hours, 4.9 hours, or 5.0 hours) from said donor. In some embodiments, the sample is isolated from the donor about 4 hours after administration of the CXCR2 agonist and CXCR4 antagonist.

In some embodiments of any of the above aspects of the invention, the CXCR2 agonist is Gro-
β T or a variant thereof. In some embodiments, the CXCR2 agonist is at least about 85% (e.g., at least about 85%) of the amino acid sequence of SEQ ID NO:2.
%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity. In some embodiments, the CXCR2 agonist is SEQ ID NO:
A peptide having about 85% to 100% sequence identity to the amino acid sequence of SEQ ID NO.
(SEQ ID NO): About 86% to about 100%, about 87% to about 99%, about 88% to about 98%,
Peptides having sequence identity of about 89% to about 97%, about 90% to about 96%, or about 91% to about 95%.
In some embodiments, the CXCR2 agonist comprises only one or more conservative amino acid substitutions (e.g., only 1-10 conservative amino acid substitutions, 1-5 conservative amino acid substitutions, or only by 1 to 3 conservative amino acid substitutions, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 conservative amino acid substitutions), SEQ ID NO. ID NO): A peptide having an amino acid sequence different from that of ID NO. 2. In some embodiments, said CXCR2
The agonist is Gro-βT. In some embodiments, the Gro-β T is not covalently modified. In some embodiments, the Gro-β T is not covalently modified with a polyalkylene glycol moiety, such as a polyethylene glycol moiety.

In some embodiments of any of the above aspects of the invention, the CXCR2 agonist is Gro-
β or a variant thereof. In some embodiments, the CXCR2 agonist is at least about 85% (e.g., about 85%, 90%, 95%, 9
The peptides may have sequence identity of 6%, 97%, 98%, 99%, or more. In some embodiments, the CXCR2 agonist is a peptide having about 85% to 100% sequence identity to the amino acid sequence of SEQ ID NO: 1, e.g. :
About 86% to about 100%, about 87% to about 99%, about 88% to about 98%, about 89% to about 97%,
Peptides having about 90% to about 96%, or about 91% to about 95% sequence identity. In some embodiments, the CXCR2 agonist comprises only one or more conservative amino acid substitutions (e.g., only 1-10 conservative amino acid substitutions, 1-5 conservative amino acid substitutions, or 1-3
SEQ ID NO: (SEQ ID NO): It is a peptide that has an amino acid sequence different from that of 1. In some embodiments, the CXCR2 agonist is Gro-β. In some embodiments, the Gro-β is not covalently modified. In some embodiments, the Gro-β is not covalently modified with a polyalkylene glycol moiety, such as a polyethylene glycol moiety.

In some embodiments, the CXCR2 agonist (e.g., Gro-β or Gro-β T, e.g., unmodified Gro-β or Gro-β T) is administered at about 50 μg/kg to about 1 mg/kg. at a dose of, for example, about 50 μg/kg, 55 μg/kg, 60 μg/kg, 65 μg/kg, 70 μg/kg, 75 μg/kg, 80 μg/kg,
85 μg/kg, 90 μg/kg, 95 μg/kg, 100 μg/kg, 105 μg/kg, 110 μg/kg, 115 μg/kg
, 120 μg/kg, 125 μg/kg, 130 μg/kg, 135 μg/kg, 140 μg/kg, 145 μg/kg, 150 μg
g/kg, 155 μg/kg, 160 μg/kg, 165 μg/kg, 170 μg/kg, 175 μg/kg, 180 μg/kg, 18
5 μg/kg, 190 μg/kg, 195 μg/kg, 200 μg/kg, 205 μg/kg, 210 μg/kg, 215 μg/kg
, 220 μg/kg, 225 μg/kg, 230 μg/kg, 235 μg/kg, 240 μg/kg, 245 μg/kg, 250 μg
g/kg, 255 μg/kg, 260 μg/kg, 265 μg/kg, 270 μg/kg, 275 μg/kg, 280 μg/kg, 28
5 μg/kg, 290 μg/kg, 295 μg/kg, 300 μg/kg, 305 μg/kg, 310 μg/kg, 315 μg/kg
, 320 μg/kg, 325 μg/kg, 330 μg/kg, 335 μg/kg, 340 μg/kg, 345 μg/kg, 350 μg
g/kg, 355 μg/kg, 360 μg/kg, 365 μg/kg, 370 μg/kg, 375 μg/kg, 380 μg/kg, 40
0 μg/kg, 405 μg/kg, 410 μg/kg, 415 μg/kg, 425 μg/kg, 430 μg/kg, 435 μg/kg
, 440 μg/kg, 445 μg/kg, 450 μg/kg, 210 μg/kg, 300 μg/kg, 400 μg/kg, 405 μ
g/kg, 410 μg/kg, 415 μg/kg, 420 μg/kg, 425 μg/kg, 430 μg/kg, 435 μg/kg, 44
0 μg/kg, 445 μg/kg, 450 μg/kg, 455 μg/kg, 460 μg/kg, 465 μg/kg, 470 μg/kg
, 475 μg/kg, 480 μg/kg, 485 μg/kg, 490 μg/kg, 495 μg/kg, 500 μg/kg, 505 μ
g/kg, 510 μg/kg, 505 μg/kg, 515 μg/kg, 520 μg/kg, 525 μg/kg, 530 μg/kg, 54
5 μg/kg, 550 μg/kg, 555 μg/kg, 560 μg/kg, 565 μg/kg, 570 μg/kg, 575 μg/kg
, 580 μg/kg, 585 μg/kg, 590 μg/kg, 595 μg/kg, 600 μg/kg, 605 μg/kg, 610 μg
g/kg, 615 μg/kg, 620 μg/kg, 625 μg/kg, 630 μg/kg, 635 μg/kg, 640 μg/kg, 64
5 μg/kg, 650 μg/kg, 655 μg/kg, 660 μg/kg, 665 μg/kg, 670 μg/kg, 675 μg/kg
, 680 μg/kg, 685 μg/kg, 690 μg/kg, 695 μg/kg, 700 μg/kg, 705 μg/kg, 710 μg
g/kg, 715 μg/kg, 720 μg/kg, 725 μg/kg, 730 μg/kg, 735 μg/kg, 740 μg/kg, 74
5 μg/kg, 750 μg/kg, 755 μg/kg, 760 μg/kg, 765 μg/kg, 770 μg/kg, 775 μg/kg
, 780 μg/kg, 785 μg/kg, 790 μg/kg, 795 μg/kg, 800 μg/kg, 805 μg/kg, 810 μg
g/kg, 815 μg/kg, 820 μg/kg, 825 μg/kg, 830 μg/kg, 835 μg/kg, 840 μg/kg, 84
5 μg/kg, 850 μg/kg, 855 μg/kg, 860 μg/kg, 865 μg/kg, 870 μg/Kg, 875 μg/kg
, 880 μg/kg, 885 μg/kg, 890 μg/kg, 895 μg/kg, 900 μg/kg, 905 μg/kg, 910 μg
g/kg, 915 μg/kg, 920 μg/kg, 925 μg/kg, 930 μg/kg, 935 μg/kg, 940 μg/kg, 94
5 μg/kg, 950 μg/kg, 955 μg/kg, 960 μg/kg, 965 μg/kg, 970 μg/kg, 975 μg/kg
, 980 μg/kg, 985 μg/kg, 990 μg/kg, 995 μg/kg, or 1,000 μg/kg to said donor. In some embodiments, the CXCR2 agonist (e.g., Gro-β or Gro-β T, e.g., unmodified Gro-β or Gro-β T) is administered at about 50 μg/kg to about 300 μg/kg.
g/kg, such as from about 100 μg/kg to about 250 μg/kg, or from about 125 μg/kg to about 225 μg/kg.
to said donor. In some embodiments, about 150 μg of the CXCR2 agonist (e.g., Gro-β or Gro-β T, e.g., unmodified Gro-β or Gro-β T)
/kg to said donor.

In another aspect, the invention relates to a method of mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor (e.g., a human donor) into the peripheral blood, the method comprising
, Gro-β T and variants thereof.
Doses from μg/kg to about 1 mg/kg (e.g., 50 μg/kg, 55 μg/kg, 60 μg/kg, 65 μg/kg, 70
μg/kg, 75 μg/kg, 80 μg/kg, 85 μg/kg, 90 μg/kg, 95 μg/kg, 100 μg/kg, 105
μg/kg, 110 μg/kg, 115 μg/kg, 120 μg/kg, 125 μg/kg, 130 μg/kg, 135 μg/kg,
140 μg/kg, 145 μg/kg, 150 μg/kg, 155 μg/kg, 160 μg/kg, 165 μg/kg, 170 μg/
kg, 175 μg/kg, 180 μg/kg, 185 μg/kg, 190 μg/kg, 195 μg/kg, 200 μg/kg, 205
μg/kg, 210 μg/kg, 215 μg/kg, 220 μg/kg, 225 μg/kg, 230 μg/kg, 235 μg/kg,
240 μg/kg, 245 μg/kg, 250 μg/kg, 255 μg/kg, 260 μg/kg, 265 μg/kg, 270 μg/
kg, 275 μg/kg, 280 μg/kg, 285 μg/kg, 290 μg/kg, 295 μg/kg, 300 μg/kg, 305
μg/kg, 310 μg/kg, 315 μg/kg, 320 μg/kg, 325 μg/kg, 330 μg/kg, 335 μg/kg,
340 μg/kg, 345 μg/kg, 350 μg/kg, 355 μg/kg, 360 μg/kg, 365 μg/kg, 370 μg/
kg, 375 μg/kg, 380 μg/kg, 400 μg/kg, 405 μg/kg, 410 μg/kg, 415 μg/kg, 425
μg/kg, 430 μg/kg, 435 μg/kg, 440 μg/kg, 445 μg/kg, 450 μg/kg, 210 μg/kg,
300 μg/kg, 400 μg/kg, 405 μg/kg, 410 μg/kg, 415 μg/kg, 420 μg/kg, 425 μg/
kg, 430 μg/kg, 435 μg/kg, 440 μg/kg, 445 μg/kg, 450 μg/kg, 455 μg/kg, 460
μg/kg, 465 μg/kg, 470 μg/kg, 475 μg/kg, 480 μg/kg, 485 μg/kg, 490 μg/kg,
495 μg/kg, 500 μg/kg, 505 μg/kg, 510 μg/kg, 505 μg/kg, 515 μg/kg, 520 μg/
kg, 525 μg/kg, 530 μg/kg, 545 μg/kg, 550 μg/kg, 555 μg/kg, 560 μg/kg, 565
μg/kg, 570 μg/kg, 575 μg/kg, 580 μg/kg, 585 μg/kg, 590 μg/kg, 595 μg/kg,
600 μg/kg, 605 μg/kg, 610 μg/kg, 615 μg/kg, 620 μg/kg, 625 μg/kg, 630 μg/
kg, 635 μg/kg, 640 μg/kg, 645 μg/kg, 650 μg/kg, 655 μg/kg, 660 μg/kg, 665
μg/kg, 670 μg/kg, 675 μg/kg, 680 μg/kg, 685 μg/kg, 690 μg/kg, 695 μg/kg,
700 μg/kg, 705 μg/kg, 710 μg/kg, 715 μg/kg, 720 μg/kg, 725 μg/kg, 730 μg/
kg, 735 μg/kg, 740 μg/kg, 745 μg/kg, 750 μg/kg, 755 μg/kg, 760 μg/kg, 765
μg/kg, 770 μg/kg, 775 μg/kg, 780 μg/kg, 785 μg/kg, 790 μg/kg, 795 μg/kg,
800 μg/kg, 805 μg/kg, 810 μg/kg, 815 μg/kg, 820 μg/kg, 825 μg/kg, 830 μg/
kg, 835 μg/kg, 840 μg/kg, 845 μg/kg, 850 μg/kg, 855 μg/kg, 860 μg/kg, 865
μg/kg, 870 μg/Kg, 875 μg/kg, 880 μg/kg, 885 μg/kg, 890 μg/kg, 895 μg/kg,
900 μg/kg, 905 μg/kg, 910 μg/kg, 915 μg/kg, 920 μg/kg, 925 μg/kg, 930 μg/
kg, 935 μg/kg, 940 μg/kg, 945 μg/kg, 950 μg/kg, 955 μg/kg, 960 μg/kg, 965
μg/kg, 970 μg/kg, 975 μg/kg, 980 μg/kg, 985 μg/kg, 990 μg/kg, 995 μg/kg,
or 1,000 μg/kg) to said donor. In some embodiments, the method further comprises administering a CXCR4 antagonist to the donor.

In some embodiments of any of the above aspects of the invention, the CXCR2 agonist (e.g.
Gro-β or Gro-β T, e.g., unmodified Gro-β or Gro-β T) from about 50 μg/kg to
at a dose of about 300 μg/kg, e.g. about 50 μg/kg, 55 μg/kg, 60 μg/kg, 65 μg/kg, 70
μg/kg, 75 μg/kg, 80 μg/kg, 85 μg/kg, 90 μg/kg, 95 μg/kg, 100 μg/kg, 105
μg/kg, 110 μg/kg, 115 μg/kg, 120 μg/kg, 125 μg/kg, 130 μg/kg, 135 μg/kg,
140 μg/kg, 145 μg/kg, 150 μg/kg, 155 μg/kg, 160 μg/kg, 165 μg/kg, 170 μg/
kg, 175 μg/kg, 180 μg/kg, 185 μg/kg, 190 μg/kg, 195 μg/kg, 200 μg/kg, 205
μg/kg, 210 μg/kg, 215 μg/kg, 220 μg/kg, 225 μg/kg, 230 μg/kg, 235 μg/kg,
240 μg/kg, 245 μg/kg, 250 μg/kg, 255 μg/kg, 260 μg/kg, 265 μg/kg, 270 μg/
kg, 275 μg/kg, 280 μg/kg, 285 μg/kg, 290 μg/kg, 295 μg/kg, or 300 μg/kg
to said donor.

In some embodiments of any of the above aspects of the invention, the CXCR2 agonist (e.g.
Gro-β or Gro-β T, e.g., unmodified Gro-β or Gro-β T) at about 100 μg/kg
at a dose of ~250 μg/kg, e.g., about 100 μg/kg, 105 μg/kg, 110 μg/kg, 115 μg/kg
, 120 μg/kg, 125 μg/kg, 130 μg/kg, 135 μg/kg, 140 μg/kg, 145 μg/kg, 150 μg
g/kg, 155 μg/kg, 160 μg/kg, 165 μg/kg, 170 μg/kg, 175 μg/kg, 180 μg/kg, 18
5 μg/kg, 190 μg/kg, 195 μg/kg, 200 μg/kg, 205 μg/kg, 210 μg/kg, 215 μg/kg
, 220 μg/kg, 225 μg/kg, 230 μg/kg, 235 μg/kg, 240 μg/kg, 245 μg/kg, or 2
A dose of 50 μg/kg is administered to the donor.

In some embodiments of any of the above aspects of the invention, the CXCR2 agonist (e.g.
Gro-β or Gro-β T, e.g., unmodified Gro-β or Gro-β T) at about 150 μg/kg
to said donor. For example, in some embodiments, the CXCR2 agonist (e.g., Gro-β or Gro-β T, e.g., unmodified Gro-β or Gro-β T) is
at a dose of about 50 μg/kg/day to about 1 mg/kg/day, e.g., about 50 μg/kg/day, 55 μg/kg/day, 60
μg/kg/day, 65 μg/kg/day, 70 μg/kg/day, 75 μg/kg/day, 80 μg/kg/day, 85 μg/kg/day,
90 μg/kg/day, 95 μg/kg/day, 100 μg/kg/day, 105 μg/kg/day, 110 μg/kg/day, 115 μg
g/kg/day, 120 μg/kg/day, 125 μg/kg/day, 130 μg/kg/day, 135 μg/kg/day, 140 μg/kg/
day, 145 μg/kg/day, 150 μg/kg/day, 155 μg/kg/day, 160 μg/kg/day, 165 μg/kg/day, 1
70 μg/kg/day, 175 μg/kg/day, 180 μg/kg/day, 185 μg/kg/day, 190 μg/kg/day, 195 μ
g/kg/day, 200 μg/kg/day, 205 μg/kg/day, 210 μg/kg/day, 215 μg/kg/day, 220 μg/kg/
day, 225 μg/kg/day, 230 μg/kg/day, 235 μg/kg/day, 240 μg/kg/day, 245 μg/kg/day, 2
50 μg/kg/day, 255 μg/kg/day, 260 μg/kg/day, 265 μg/kg/day, 270 μg/kg/day, 275 μg
g/kg/day, 280 μg/kg/day, 285 μg/kg/day, 290 μg/kg/day, 295 μg/kg/day, 300 μg/kg/
day, 305 μg/kg/day, 310 μg/kg/day, 315 μg/kg/day, 320 μg/kg/day, 325 μg/kg/day, 3
30 μg/kg/day, 335 μg/kg/day, 340 μg/kg/day, 345 μg/kg/day, 350 μg/kg/day, 355 μg/kg/day
g/kg/day, 360 μg/kg/day, 365 μg/kg/day, 370 μg/kg/day, 375 μg/kg/day, 380 μg/kg/
day, 400 μg/kg/day, 405 μg/kg/day, 410 μg/kg/day, 415 μg/kg/day, 425 μg/kg/day, 4
30 μg/kg/day, 435 μg/kg/day, 440 μg/kg/day, 445 μg/kg/day, 450 μg/kg/day, 210 μg
g/kg/day, 300 μg/kg/day, 400 μg/kg/day, 405 μg/kg/day, 410 μg/kg/day, 415 μg/kg/
day, 420 μg/kg/day, 425 μg/kg/day, 430 μg/kg/day, 435 μg/kg/day, 440 μg/kg/day, 4
45 μg/kg/day, 450 μg/kg/day, 455 μg/kg/day, 460 μg/kg/day, 465 μg/kg/day, 470 μg
g/kg/day, 475 μg/kg/day, 480 μg/kg/day, 485 μg/kg/day, 490 μg/kg/day, 495 μg/kg/
day, 500 μg/kg/day, 505 μg/kg/day, 510 μg/kg/day, 505 μg/kg/day, 515 μg/kg/day, 5
20 μg/kg/day, 525 μg/kg/day, 530 μg/kg/day, 545 μg/kg/day, 550 μg/kg/day, 555 μg
g/kg/day, 560 μg/kg/day, 565 μg/kg/day, 570 μg/kg/day, 575 μg/kg/day, 580 μg/kg/
day, 585 μg/kg/day, 590 μg/kg/day, 595 μg/kg/day, 600 μg/kg/day, 605 μg/kg/day, 6
10 μg/kg/day, 615 μg/kg/day, 620 μg/kg/day, 625 μg/kg/day, 630 μg/kg/day, 635 μg
g/kg/day, 640 μg/kg/day, 645 μg/kg/day, 650 μg/kg/day, 655 μg/kg/day, 660 μg/kg/
day, 665 μg/kg/day, 670 μg/kg/day, 675 μg/kg/day, 680 μg/kg/day, 685 μg/kg/day, 6
90 μg/kg/day, 695 μg/kg/day, 700 μg/kg/day, 705 μg/kg/day, 710 μg/kg/day, 715 μg
g/kg/day, 720 μg/kg/day, 725 μg/kg/day, 730 μg/kg/day, 735 μg/kg/day, 740 μg/kg/
day, 745 μg/kg/day, 750 μg/kg/day, 755 μg/kg/day, 760 μg/kg/day, 765 μg/kg/day, 7
70 μg/kg/day, 775 μg/kg/day, 780 μg/kg/day, 785 μg/kg/day, 790 μg/kg/day, 795 μg
g/kg/day, 800 μg/kg/day, 805 μg/kg/day, 810 μg/kg/day, 815 μg/kg/day, 820 μg/kg/
day, 825 μg/kg/day, 830 μg/kg/day, 835 μg/kg/day, 840 μg/kg/day, 845 μg/kg/day, 8
50 μg/kg/day, 855 μg/kg/day, 860 μg/kg/day, 865 μg/kg/day, 870 μg/kg/day, 875 μg/kg/day
g/kg/day, 880 μg/kg/day, 885 μg/kg/day, 890 μg/kg/day, 895 μg/kg/day, 900 μg/kg/
day, 905 μg/kg/day, 910 μg/kg/day, 915 μg/kg/day, 920 μg/kg/day, 925 μg/kg/day, 9
30 μg/kg/day, 935 μg/kg/day, 940 μg/kg/day, 945 μg/kg/day, 950 μg/kg/day, 955 μg
g/kg/day, 960 μg/kg/day, 965 μg/kg/day, 970 μg/kg/day, 975 μg/kg/day, 980 μg/kg/
at a dose of 985 μg/kg/day, 990 μg/kg/day, 995 μg/kg/day, or 1,000 μg/kg/day,
administered to said donor. In some embodiments, the CXCR2 agonist (e.g., Gro-β
or Gro-β T, e.g., unmodified Gro-β or Gro-β T) from about 50 μg/kg/day to about 3
00 μg/kg/day, for example, from about 100 μg/kg/day to about 250 μg/kg/day, or about 125 μg/kg
or about 125 μg/kg/day to about 175 μg/kg/day to said donor. In some embodiments, the CXCR2 agonist (e.g., Gro-β or Gro-β T
, eg, unmodified Gro-β or Gro-β T) at a dose of about 150 μg/kg/day to said donor. In some embodiments, the CXCR2 agonist may be administered in a single dose. In some embodiments, the CXCR2 agonist may be administered in multiple doses.

In some embodiments, a human equivalent dose (HED) may be derived from animal dose data using a conversion factor. For example, Nair and Jacob, JBasic Cli
n Pharma. (2016)7:27-31 discloses a method for extrapolating doses between species. For example, in one non-limiting example, the HED may be derived from the rhesus dose by multiplying the rhesus dose by about 0.324.

In some embodiments of any of the above aspects, the CXCR2 agonist (e.g., Gro-
β or Gro-β T, eg, unmodified Gro-β or Gro-β T) is administered intravenously to said donor.

In some embodiments of any of the above aspects of the invention, the CXCR4 antagonist is
Formula (I)
Z -Linker- Z' (I)
or a pharmaceutically acceptable salt thereof, where Z is:
(i) is a cyclic polyamine containing from 9 to 32 ring members, where 2 to 8 ring members are
are nitrogen atoms separated from each other by two or more carbon atoms; or
(ii) An amine represented by formula (IA)

where A comprises a monocyclic or bicyclic fused ring system containing at least one nitrogen atom, and B is H or a substituent consisting of 1 to 20 atoms;
and where Z' is:
(i) is a cyclic polyamine containing from 9 to 32 ring members, where 2 to 8 ring members are
are nitrogen atoms separated from each other by two or more carbon atoms;
(ii) An amine represented by formula (IB)

where A' comprises a monocyclic or bicyclic fused ring system containing at least one nitrogen atom, and B' is H or a substituent consisting of 1 to 20 atoms; or
(iii) Substituent represented by formula (IC)
-N(R)-(CR ₂ ) _n -X (IC)
where each R is independently H or _C1 - _C6 alkyl, n is 1 or 2, and X is an aryl or heteroaryl group or a mercaptan;
wherein said linker is a bond, an optionally substituted C ₁ -C ₆ alkylene, an optionally substituted C ₁ -C ₆ heteroalkylene, an optionally substituted C ₂ -C ₆ alkenylene, optionally substituted _C2 - _C6 heteroalkenylene, optionally substituted _C2 - _C6 alkynylene, optionally substituted _C2 - _C6 heteroalkynylene, optionally substituted optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene, or optionally substituted heteroarylene.

In some embodiments, Z and Z' are each independently a cyclic polyamine containing 9 to 32 ring members, and 2 to 8 of the 9 to 32 ring members are Nitrogen atoms separated from each other by two or more carbon atoms. Z and Z' may be the same substituent. In some embodiments, Z and/or Z' is a cyclic polyamine containing 10-24 ring members, such as a cyclic polyamine containing 14 ring members. In some embodiments, Z includes 4 nitrogen atoms. Z and/or Z' may be, for example, 1,4,8,11-tetraazocyclotetradecane.

In some embodiments, the linker is represented by the formula (ID),

where Ring D is an optionally substituted aryl group, an optionally substituted heteroaryl group, an optionally substituted cycloalkyl group, or an optionally substituted heterocycloalkyl group is; and
X and Y are each independently optionally substituted _C1 - _C6 alkylene, optionally substituted _C1 - _C6 heteroalkylene, optionally substituted _C2 -C ₆ alkenylene, optionally substituted _C2 - _C6 heteroalkenylene, optionally substituted _C2 - _C6 alkynylene, or optionally substituted _C2 - _C6 heteroalkynylene .

In some embodiments, the linker is represented by the formula (IE),

where Ring D is an optionally substituted aryl group, an optionally substituted heteroaryl group, an optionally substituted cycloalkyl group, or an optionally substituted heterocycloalkyl group is; and
X and Y are each independently optionally substituted _C1 - _C6 alkylene, optionally substituted _C1 - _C6 heteroalkylene, optionally substituted _C2 -C ₆ alkenylene, optionally substituted _C2 - _C6 heteroalkenylene, optionally substituted _C2 - _C6 alkynylene, or optionally substituted _C2 - _C6 heteroalkynylene .

In some embodiments, X and Y are each independently optionally substituted _C1 - _C6 alkylene. In some embodiments, X and Y are the same substituents, such as the same alkylene substituents (eg, methylene, ethylene, propylene, or butylene substituents).

In some embodiments, the CXCR4 antagonist is plerixafor or a pharmaceutically acceptable salt thereof. In some embodiments, the CXCR4 antagonist (e.g.
Plerixafor or a pharmaceutically acceptable salt thereof) is administered subcutaneously to said donor. In some embodiments, the CXCR4 antagonist (e.g., plerixafor or a pharmaceutically acceptable salt thereof) is administered at a dose of about 50 μg/kg to about 500 μg/kg, e.g., about 50 μg/kg, 55 μg/
kg, 60 μg/kg, 65 μg/kg, 70 μg/kg, 75 μg/kg, 80 μg/kg, 85 μg/kg, 90 μg/kg,
95 μg/kg, 100 μg/kg, 105 μg/kg, 110 μg/kg, 115 μg/kg, 120 μg/kg, 125 μg/
kg, 130 μg/kg, 135 μg/kg, 140 μg/kg, 145 μg/kg, 150 μg/kg, 155 μg/kg, 160
μg/kg, 165 μg/kg, 170 μg/kg, 175 μg/kg, 180 μg/kg, 185 μg/kg, 190 μg/kg,
195 μg/kg, 200 μg/kg, 205 μg/kg, 210 μg/kg, 215 μg/kg, 220 μg/kg, 225 μg/
kg, 230 μg/kg, 235 μg/kg, 240 μg/kg, 245 μg/kg, 250 μg/kg, 255 μg/kg, 260
μg/kg, 265 μg/kg, 270 μg/kg, 275 μg/kg, 280 μg/kg, 285 μg/kg, 290 μg/kg,
295 μg/kg, 300 μg/kg, 305 μg/kg, 310 μg/kg, 315 μg/kg, 320 μg/kg, 325 μg/
kg, 330 μg/kg, 335 μg/kg, 340 μg/kg, 345 μg/kg, 350 μg/kg, 355 μg/kg, 360
μg/kg, 365 μg/kg, 370 μg/kg, 375 μg/kg, 380 μg/kg, 385 μg/kg, 390 μg/kg,
395 μg/kg, 400 μg/kg, 405 μg/kg, 410 μg/kg, 415 μg/kg, 420 μg/kg, 425 μg/
kg, 430 μg/kg, 435 μg/kg, 440 μg/kg, 445 μg/kg, 450 μg/kg, 455 μg/kg, 460
μg/kg, 465 μg/kg, 470 μg/kg, 475 μg/kg, 480 μg/kg, 485 μg/kg, 490 μg/kg,
A dose of 495 μg/kg, or 500 μg/kg is administered to the donor. In some embodiments, the CXCR4 antagonist (e.g., plerixafor or a pharmaceutically acceptable salt thereof)
is administered to the donor at a dose of about 200 μg/kg to about 300 μg/kg, such as a dose of about 240 μg/kg.

For example, in some embodiments, the CXCR4 antagonist (e.g., plerixafor or a pharmaceutically acceptable salt thereof) is administered at a dose of about 50 μg/kg/day to about 500 μg/kg/day, e.g. , about 50 μg/kg/day, 55 μg/kg/day, 60 μg/kg/day, 65 μg/kg/day, 70 μg/kg/day, 75
μg/kg/day, 80 μg/kg/day, 85 μg/kg/day, 90 μg/kg/day, 95 μg/kg/day, 100 μg/kg/day
, 105 μg/kg/day, 110 μg/kg/day, 115 μg/kg/day, 120 μg/kg/day, 125 μg/kg/day, 130
μg/kg/day, 135 μg/kg/day, 140 μg/kg/day, 145 μg/kg/day, 150 μg/kg/day, 155 μg/
kg/day, 160 μg/kg/day, 165 μg/kg/day, 170 μg/kg/day, 175 μg/kg/day, 180 μg/kg/day
, 185 μg/kg/day, 190 μg/kg/day, 195 μg/kg/day, 200 μg/kg/day, 205 μg/kg/day, 210
μg/kg/day, 215 μg/kg/day, 220 μg/kg/day, 225 μg/kg/day, 230 μg/kg/day, 235 μg/day
kg/day, 240 μg/kg/day, 245 μg/kg/day, 250 μg/kg/day, 255 μg/kg/day, 260 μg/kg/day
, 265 μg/kg/day, 270 μg/kg/day, 275 μg/kg/day, 280 μg/kg/day, 285 μg/kg/day, 290
μg/kg/day, 295 μg/kg/day, 300 μg/kg/day, 305 μg/kg/day, 310 μg/kg/day, 315 μg/day
kg/day, 320 μg/kg/day, 325 μg/kg/day, 330 μg/kg/day, 335 μg/kg/day, 340 μg/kg/day
, 345 μg/kg/day, 350 μg/kg/day, 355 μg/kg/day, 360 μg/kg/day, 365 μg/kg/day, 370
μg/kg/day, 375 μg/kg/day, 380 μg/kg/day, 385 μg/kg/day, 390 μg/kg/day, 395 μg/
kg/day, 400 μg/kg/day, 405 μg/kg/day, 410 μg/kg/day, 415 μg/kg/day, 420 μg/kg/day
, 425 μg/kg/day, 430 μg/kg/day, 435 μg/kg/day, 440 μg/kg/day, 445 μg/kg/day, 450
μg/kg/day, 455 μg/kg/day, 460 μg/kg/day, 465 μg/kg/day, 470 μg/kg/day, 475 μg/
kg/day, 480 μg/kg/day, 485 μg/kg/day, 490 μg/kg/day, 495 μg/kg/day, or 500 μg/day
kg/day to said donor. In some embodiments, the CXCR4 antagonist (e.g., plerixafor or a pharmaceutically acceptable salt thereof) is administered at about 200 μg/kg/day to about 3
00 μg/kg/day, for example at a dose of about 240 μg/kg/day. In some embodiments, the CXCR4 antagonist may be administered in a single dose. In other embodiments, the CXCR4 antagonist may be administered in multiple doses.

In some embodiments of any of the above aspects of the invention, said CXCR2 agonist and said C
An XCR4 antagonist is simultaneously administered to said donor. In some embodiments, the CX
The CXCR4 antagonist is administered to the donor prior to administration of the CR2 agonist. In some embodiments, from about 1 minute to about 180 minutes, such as from about 15 minutes to about 180 minutes, before administration of said CXCR2 agonist.
Approx. 180 minutes, Approx. 30 minutes to approx. 180 minutes, Approx. 40 minutes to approx. 160 minutes, Approx. 50 minutes to approx. 150 minutes, Approx. 60 minutes to approx. 140 minutes, Approx. 7
0 minutes to about 130 minutes, about 60 minutes to about 120 minutes, about 70 minutes to about 110 minutes, or about 80 minutes to about 100 minutes (e.g., about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes, about 60
minutes, about 65 minutes, about 70 minutes, about 75 minutes, about 80 minutes, about 85 minutes, about 90 minutes, about 95 minutes, about 100 minutes, about 105 minutes, about
110 minutes, about 115 minutes, about 120 minutes, about 125 minutes, about 130 minutes, about 135 minutes, about 140 minutes, about 145 minutes, about 150 minutes,
The CXCR4 antagonist is administered to the donor at about 155 minutes, about 160 minutes, about 165 minutes, about 170 minutes, about 175 minutes, or about 180 minutes. In some embodiments, about 30 minutes to about 60 minutes before administration of said CXCR2 agonist (e.g., about 30 minutes, about 35 minutes, about 40 minutes before administration of said CXCR2 agonist)
The CXCR4 antagonist is administered to the donor at about 45 minutes, about 45 minutes, about 50 minutes, about 55 minutes, or about 60 minutes). In some embodiments, about 45 minutes before administration of said CXCR2 agonist, said C
An XCR4 antagonist is administered to said donor.

In a further aspect, the invention relates to a pharmaceutical composition comprising a population of hematopoietic stem cells or progeny thereof isolated from a mammalian donor (e.g., a human donor), wherein leukocytes in said population The ratio of CD34 ⁺ cells to CD34+ cells is about 0.0008 to about 0.0021. In some embodiments, the ratio of CD34 ⁺ cells to white blood cells is about 0.00080, 0.00081, 0.00082, 0.0008
3, 0.00084, 0.00085, 0.00086, 0.00087, 0.00088, 0.00089, 0.00090, 0.00091, 0.000
92, 0.00093, 0.00094, 0.00095, 0.00096, 0.00097, 0.00098, 0.00099, 0.00100, 0.00
101, 0.00102, 0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108, 0.00109, 0.0
0110, 0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117, 0.00118, 0.
00119, 0.00120, 0.00121, 0.00122, 0.00123, 0.00124, 0.00125, 0.00126, 0.00127, 0
.00128, 0.00129, 0.00130, 0.00131, 0.00132, 0.00133, 0.00134, 0.00135, 0.00136,
0.00137, 0.00138, 0.00139, 0.00140, 0.00141, 0.00142, 0.00143, 0.00144, 0.00145,
0.00146, 0.00147, 0.00148, 0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154
, 0.00155, 0.00156, 0.00157, 0.00158, 0.00159, 0.00160, 0.00161, 0.00162, 0.0016
3, 0.00164, 0.00165, 0.00166, 0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.001
72, 0.00173, 0.00174, 0.00175, 0.00176, 0.00178, 0.00179, 0.00180, 0.00181, 0.00
182, 0.00183, 0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189, 0.00190, 0.0
0191, 0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197, 0.00198, 0.00199, 0.
00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205, 0.00206, 0.00207, 0.00208, 0
.00209, 0.00210, 0.00211, 0.00212, 0.00213, 0.00214, 0.00215, 0.00216, 0.00217,
May be 0.00218, 0.00219, 0.00220, 0.00221, 0.00222, 0.00223, 0.00224, or 0.00225. In some embodiments, the ratio of CD34 ⁺ cells to white blood cells is about 0.0010
~about 0.0018, for example, about 0.00100, 0.00101, 0.00102, 0.00103, 0.00104, 0.00105, 0.00
106, 0.00107, 0.00108, 0.00109, 0.00110, 0.00111, 0.00112, 0.00113, 0.00114, 0.0
0115, 0.00116, 0.00117, 0.00118, 0.00119, 0.00120, 0.00121, 0.00122, 0.00123, 0.
00124, 0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00132, 0
.00133, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.00141,
0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.00150,
0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158, 0.00159
, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164, 0.00165, 0.00166, 0.00167, 0.0016
8, 0.00169, 0.00170, 0.00171, 0.00172, 0.00173, 0.00174, 0.00175, 0.00176, 0.001
The ratio of CD34 ⁺ cells to white blood cells is 78, 0.00179, or 0.00180. In some embodiments, the ratio of CD34 ⁺ cells to white blood cells is about 0.0014.

In a further aspect, the present invention relates to a pharmaceutical composition comprising a population of hematopoietic stem cells or their progeny isolated from a mammalian donor (e.g., a human donor), wherein a preferred population of hematopoietic stem cells or progeny thereof is The ratio of CD34 ⁺ cells to mesospheres is about 0.0018 to about 0.0058. In some embodiments, the ratio of CD34 ⁺ cells to neutrophils is about 0.00180, 0.00181, 0.00182, 0.0018
3, 0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189, 0.00190, 0.00191, 0.001
92, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197, 0.00198, 0.00199, 0.00200, 0.00
201, 0.00202, 0.00203, 0.00204, 0.00205, 0.00206, 0.00207, 0.00208, 0.00209, 0.0
0210, 0.00211, 0.00212, 0.00213, 0.00214, 0.00215, 0.00216, 0.00217, 0.00218, 0.
00219, 0.00220, 0.00221, 0.00222, 0.00223, 0.00224, 0.00225, 0.00226, 0.00227, 0
.00228, 0.00229, 0.00230, 0.00231, 0.00232, 0.00233, 0.00234, 0.00235, 0.00236,
0.00237, 0.00238, 0.00239, 0.00240, 0.00241, 0.00242, 0.00243, 0.00244, 0.00245,
0.00246, 0.00247, 0.00248, 0.00249, 0.00250, 0.00251, 0.00252, 0.00253, 0.00254
, 0.00255, 0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.00261, 0.00262, 0.0026
3, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.002
72, 0.00273, 0.00274, 0.00275, 0.00276, 0.00277, 0.00278, 0.00279, 0.00280, 0.00
281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288, 0.00289, 0.0
0290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.00297, 0.00298, 0.
00299, 0.00300, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0
.00307, 0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313, 0.00314, 0.00315,
0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324,
0.00325, 0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333
, 0.00334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339, 0.00340, 0.00341, 0.0034
2, 0.00343, 0.00344, 0.00345, 0.00346, 0.00347, 0.00348, 0.00349, 0.00350, 0.003
51, 0.00352, 0.00353, 0.00354, 0.00355, 0.00356, 0.00357, 0.00358, 0.00359, 0.00
360, 0.00361, 0.00362, 0.00363, 0.00364, 0.00365, 0.00366, 0.00367, 0.00368, 0.0
0369, 0.00370, 0.00371, 0.00372, 0.00373, 0.00374, 0.00375, 0.00376, 0.00377, 0.
00378, 0.00379, 0.00380, 0.00381, 0.00382, 0.00383, 0.00384, 0.00385, 0.00386, 0
.00387, 0.00388, 0.00389, 0.00390, 0.00391, 0.00392, 0.00393, 0.00394, 0.00395,
0.00396, 0.00397, 0.00398, 0.00399, 0.00400, 0.00401, 0.00402, 0.00403, 0.00404,
0.00405, 0.00406, 0.00407, 0.00408, 0.00409, 0.00410, 0.00411, 0.00412, 0.00413
, 0.00414, 0.00415, 0.00416, 0.00417, 0.00418, 0.00419, 0.00420, 0.00421, 0.0042
2, 0.00423, 0.00424, 0.00425, 0.00426, 0.00427, 0.00428, 0.00429, 0.00430, 0.004
31, 0.00432, 0.00433, 0.00434, 0.00435, 0.00436, 0.00437, 0.00438, 0.00439, 0.00
440, 0.00441, 0.00442, 0.00443, 0.00444, 0.00445, 0.00446, 0.00447, 0.00448, 0.0
0449, 0.00450, 0.00451, 0.00452, 0.00453, 0.00454, 0.00455, 0.00456, 0.00457, 0.
00458, 0.00459, 0.00460, 0.00461, 0.00462, 0.00463, 0.00464, 0.00465, 0.00466, 0
.00467, 0.00468, 0.00469, 0.00470, 0.00471, 0.00472, 0.00473, 0.00474, 0.00475,
0.00476, 0.00477, 0.00478, 0.00479, 0.00480, 0.00481, 0.00482, 0.00483, 0.00484,
0.00485, 0.00486, 0.00487, 0.00488, 0.00489, 0.00490, 0.00491, 0.00492, 0.00493
, 0.00494, 0.00495, 0.00496, 0.00497, 0.00498, 0.00499, 0.00500, 0.00501, 0.0050
2, 0.00503, 0.00504, 0.00505, 0.00506, 0.00507, 0.00508, 0.00509, 0.00510, 0.005
11, 0.00512, 0.00513, 0.00514, 0.00515, 0.00516, 0.00517, 0.00518, 0.00519, 0.00
520, 0.00521, 0.00522, 0.00523, 0.00524, 0.00525, 0.00526, 0.00527, 0.00528, 0.0
0529, 0.00530, 0.00531, 0.00532, 0.00533, 0.00534, 0.00535, 0.00536, 0.00537, 0.
00538, 0.00539, 0.00540, 0.00541, 0.00542, 0.00543, 0.00544, 0.00545, 0.00546, 0
.00547, 0.00548, 0.00549, 0.00550, 0.00551, 0.00552, 0.00553, 0.00554, 0.00555,
0.00556, 0.00557, 0.00558, 0.00559, 0.00560, 0.00561, 0.00562, 0.00563, 0.00564,
0.00565, 0.00566, 0.00567, 0.00568, 0.00569, 0.00570, 0.00571, 0.00572, 0.00573
, 0.00574, 0.00575, 0.00576, 0.00577, 0.00578, 0.00579, or 0.00580. In some embodiments, the ratio of CD34 ⁺ cells to neutrophils is about 0.0026 to about 0.004
6, for example, approximately 0.00260, 0.00261, 0.00262, 0.00263, 0.00264, 0.00265, 0.00266, 0.00
267, 0.00268, 0.00269, 0.00270, 0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.0
0276, 0.00277, 0.00278, 0.00279, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.
00285, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0
.00294, 0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.00300, 0.00301,
0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310,
0.00311, 0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319
, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325, 0.00326, 0.00327, 0.0032
8, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.003
37, 0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343, 0.00344, 0.00345, 0.00
346, 0.00347, 0.00348, 0.00349, 0.00350, 0.00351, 0.00352, 0.00353, 0.00354, 0.0
0355, 0.00356, 0.00357, 0.00358, 0.00359, 0.00360, 0.00361, 0.00362, 0.00363, 0.
00364, 0.00365, 0.00366, 0.00367, 0.00368, 0.00369, 0.00370, 0.00371, 0.00372, 0
.00373, 0.00374, 0.00375, 0.00376, 0.00377, 0.00378, 0.00379, 0.00380, 0.00381,
0.00382, 0.00383, 0.00384, 0.00385, 0.00386, 0.00387, 0.00388, 0.00389, 0.00390,
0.00391, 0.00392, 0.00393, 0.00394, 0.00395, 0.00396, 0.00397, 0.00398, 0.00399
, 0.00400, 0.00401, 0.00402, 0.00403, 0.00404, 0.00405, 0.00406, 0.00407, 0.0040
8, 0.00409, 0.00410, 0.00411, 0.00412, 0.00413, 0.00414, 0.00415, 0.00416, 0.004
17, 0.00418, 0.00419, 0.00420, 0.00421, 0.00422, 0.00423, 0.00424, 0.00425, 0.00
426, 0.00427, 0.00428, 0.00429, 0.00430, 0.00431, 0.00432, 0.00433, 0.00434, 0.0
0435, 0.00436, 0.00437, 0.00438, 0.00439, 0.00440, 0.00441, 0.00442, 0.00443, 0.
00444, 0.00445, 0.00446, 0.00447, 0.00448, 0.00449, 0.00450, 0.00451, 0.00452, 0
The ratio of CD34 ⁺ cells to neutrophils is .00453, 0.00454, 0.00455, 0.00456, 0.00457, 0.00458, 0.00459, or 0.00460. In some embodiments, the ratio of CD34 ⁺ cells to neutrophils is about 0.0036.

In another aspect, the present invention relates to a pharmaceutical composition comprising a population of hematopoietic stem cells or their progeny isolated from a mammalian donor (e.g., a human donor), wherein lymphocytes in said population The ratio of CD34 ⁺ cells to spheres is about 0.0021 to about 0.0094. In some embodiments, the ratio of CD34 ⁺ cells to lymphocytes is about 0.00210, 0.00211, 0.00212, 0.00
213, 0.00214, 0.00215, 0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.0
0222, 0.00223, 0.00224, 0.00225, 0.00226, 0.00227, 0.00228, 0.00229, 0.00230, 0.
00231, 0.00232, 0.00233, 0.00234, 0.00235, 0.00236, 0.00237, 0.00238, 0.00239, 0
.00240, 0.00241, 0.00242, 0.00243, 0.00244, 0.00245, 0.00246, 0.00247, 0.00248,
0.00249, 0.00250, 0.00251, 0.00252, 0.00253, 0.00254, 0.00255, 0.00256, 0.00257,
0.00258, 0.00259, 0.00260, 0.00261, 0.00262, 0.00263, 0.00264, 0.00265, 0.00266
, 0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.00272, 0.00273, 0.00274, 0.0027
5, 0.00276, 0.00277, 0.00278, 0.00279, 0.00280, 0.00281, 0.00282, 0.00283, 0.002
84, 0.00285, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00
293, 0.00294, 0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.00300, 0.0
0301, 0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308, 0.00309, 0.
00310, 0.00311, 0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0
.00319, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325, 0.00326, 0.00327,
0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00335, 0.00336,
0.00337, 0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343, 0.00344, 0.00345
, 0.00346, 0.00347, 0.00348, 0.00349, 0.00350, 0.00351, 0.00352, 0.00353, 0.0035
4, 0.00355, 0.00356, 0.00357, 0.00358, 0.00359, 0.00360, 0.00361, 0.00362, 0.003
63, 0.00364, 0.00365, 0.00366, 0.00367, 0.00368, 0.00369, 0.00370, 0.00371, 0.00
372, 0.00373, 0.00374, 0.00375, 0.00376, 0.00377, 0.00378, 0.00379, 0.00380, 0.0
0381, 0.00382, 0.00383, 0.00384, 0.00385, 0.00386, 0.00387, 0.00388, 0.00389, 0.
00390, 0.00391, 0.00392, 0.00393, 0.00394, 0.00395, 0.00396, 0.00397, 0.00398, 0
.00399, 0.00400, 0.00401, 0.00402, 0.00403, 0.00404, 0.00405, 0.00406, 0.00407,
0.00408, 0.00409, 0.00410, 0.00411, 0.00412, 0.00413, 0.00414, 0.00415, 0.00416,
0.00417, 0.00418, 0.00419, 0.00420, 0.00421, 0.00422, 0.00423, 0.00424, 0.00425
, 0.00426, 0.00427, 0.00428, 0.00429, 0.00430, 0.00431, 0.00432, 0.00433, 0.0043
4, 0.00435, 0.00436, 0.00437, 0.00438, 0.00439, 0.00440, 0.00441, 0.00442, 0.004
43, 0.00444, 0.00445, 0.00446, 0.00447, 0.00448, 0.00449, 0.00450, 0.00451, 0.00
452, 0.00453, 0.00454, 0.00455, 0.00456, 0.00457, 0.00458, 0.00459, 0.00460, 0.0
0461, 0.00462, 0.00463, 0.00464, 0.00465, 0.00466, 0.00467, 0.00468, 0.00469, 0.
00470, 0.00471, 0.00472, 0.00473, 0.00474, 0.00475, 0.00476, 0.00477, 0.00478, 0
.00479, 0.00480, 0.00481, 0.00482, 0.00483, 0.00484, 0.00485, 0.00486, 0.00487,
0.00488, 0.00489, 0.00490, 0.00491, 0.00492, 0.00493, 0.00494, 0.00495, 0.00496,
0.00497, 0.00498, 0.00499, 0.00500, 0.00501, 0.00502, 0.00503, 0.00504, 0.00505
, 0.00506, 0.00507, 0.00508, 0.00509, 0.00510, 0.00511, 0.00512, 0.00513, 0.0051
4, 0.00515, 0.00516, 0.00517, 0.00518, 0.00519, 0.00520, 0.00521, 0.00522, 0.005
23, 0.00524, 0.00525, 0.00526, 0.00527, 0.00528, 0.00529, 0.00530, 0.00531, 0.00
532, 0.00533, 0.00534, 0.00535, 0.00536, 0.00537, 0.00538, 0.00539, 0.00540, 0.0
0541, 0.00542, 0.00543, 0.00544, 0.00545, 0.00546, 0.00547, 0.00548, 0.00549, 0.
00550, 0.00551, 0.00552, 0.00553, 0.00554, 0.00555, 0.00556, 0.00557, 0.00558, 0
.00559, 0.00560, 0.00561, 0.00562, 0.00563, 0.00564, 0.00565, 0.00566, 0.00567,
0.00568, 0.00569, 0.00570, 0.00571, 0.00572, 0.00573, 0.00574, 0.00575, 0.00576,
0.00577, 0.00578, 0.00579, 0.00580, 0.00581, 0.00582, 0.00583, 0.00584, 0.00585
, 0.00586, 0.00587, 0.00588, 0.00589, 0.00590, 0.00591, 0.00592, 0.00593, 0.0059
4, 0.00595, 0.00596, 0.00597, 0.00598, 0.00599, 0.00600, 0.00601, 0.00602, 0.006
03, 0.00604, 0.00605, 0.00606, 0.00607, 0.00608, 0.00609, 0.00610, 0.00611, 0.00
612, 0.00613, 0.00614, 0.00615, 0.00616, 0.00617, 0.00618, 0.00619, 0.00620, 0.0
0621, 0.00622, 0.00623, 0.00624, 0.00625, 0.00626, 0.00627, 0.00628, 0.00629, 0.
00630, 0.00631, 0.00632, 0.00633, 0.00634, 0.00635, 0.00636, 0.00637, 0.00638, 0
.00639, 0.00640, 0.00641, 0.00642, 0.00643, 0.00644, 0.00645, 0.00646, 0.00647,
0.00648, 0.00649, 0.00650, 0.00651, 0.00652, 0.00653, 0.00654, 0.00655, 0.00656,
0.00657, 0.00658, 0.00659, 0.00660, 0.00661, 0.00662, 0.00663, 0.00664, 0.00665
, 0.00666, 0.00667, 0.00668, 0.00669, 0.00670, 0.00671, 0.00672, 0.00673, 0.0067
4, 0.00675, 0.00676, 0.00677, 0.00678, 0.00679, 0.00680, 0.00681, 0.00682, 0.006
83, 0.00684, 0.00685, 0.00686, 0.00687, 0.00688, 0.00689, 0.00690, 0.00691, 0.00
692, 0.00693, 0.00694, 0.00695, 0.00696, 0.00697, 0.00698, 0.00699, 0.00700, 0.0
0701, 0.00702, 0.00703, 0.00704, 0.00705, 0.00706, 0.00707, 0.00708, 0.00709, 0.
00710, 0.00711, 0.00712, 0.00713, 0.00714, 0.00715, 0.00716, 0.00717, 0.00718, 0
.00719, 0.00720, 0.00721, 0.00722, 0.00723, 0.00724, 0.00725, 0.00726, 0.00727,
0.00728, 0.00729, 0.00730, 0.00731, 0.00732, 0.00733, 0.00734, 0.00735, 0.00736,
0.00737, 0.00738, 0.00739, 0.00740, 0.00741, 0.00742, 0.00743, 0.00744, 0.00745
, 0.00746, 0.00747, 0.00748, 0.00749, 0.00750, 0.00751, 0.00752, 0.00753, 0.0075
4, 0.00755, 0.00756, 0.00757, 0.00758, 0.00759, 0.00760, 0.00761, 0.00762, 0.007
63, 0.00764, 0.00765, 0.00766, 0.00767, 0.00768, 0.00769, 0.00770, 0.00771, 0.00
772, 0.00773, 0.00774, 0.00775, 0.00776, 0.00777, 0.00778, 0.00779, 0.00780, 0.0
0781, 0.00782, 0.00783, 0.00784, 0.00785, 0.00786, 0.00787, 0.00788, 0.00789, 0.
00790, 0.00791, 0.00792, 0.00793, 0.00794, 0.00795, 0.00796, 0.00797, 0.00798, 0
.00799, 0.00800, 0.00801, 0.00802, 0.00803, 0.00804, 0.00805, 0.00806, 0.00807,
0.00808, 0.00809, 0.00810, 0.00811, 0.00812, 0.00813, 0.00814, 0.00815, 0.00816,
0.00817, 0.00818, 0.00819, 0.00820, 0.00821, 0.00822, 0.00823, 0.00824, 0.00825
, 0.00826, 0.00827, 0.00828, 0.00829, 0.00830, 0.00831, 0.00832, 0.00833, 0.0083
4, 0.00835, 0.00836, 0.00837, 0.00838, 0.00839, 0.00840, 0.00841, 0.00842, 0.008
43, 0.00844, 0.00845, 0.00846, 0.00847, 0.00848, 0.00849, 0.00850, 0.00851, 0.00
852, 0.00853, 0.00854, 0.00855, 0.00856, 0.00857, 0.00858, 0.00859, 0.00860, 0.0
0861, 0.00862, 0.00863, 0.00864, 0.00865, 0.00866, 0.00867, 0.00868, 0.00869, 0.
00870, 0.00871, 0.00872, 0.00873, 0.00874, 0.00875, 0.00876, 0.00877, 0.00878, 0
.00879, 0.00880, 0.00881, 0.00882, 0.00883, 0.00884, 0.00885, 0.00886, 0.00887,
0.00888, 0.00889, 0.00890, 0.00891, 0.00892, 0.00893, 0.00894, 0.00895, 0.00896,
0.00897, 0.00898, 0.00899, 0.00900, 0.00901, 0.00902, 0.00903, 0.00904, 0.00905
, 0.00906, 0.00907, 0.00908, 0.00909, 0.00910, 0.00911, 0.00912, 0.00913, 0.0091
4, 0.00915, 0.00916, 0.00917, 0.00918, 0.00919, 0.00920, 0.00921, 0.00922, 0.009
23, 0.00924, 0.00925, 0.00926, 0.00927, 0.00928, 0.00929, 0.00930, 0.00931, 0.00
932, 0.00933, 0.00934, 0.00935, 0.00936, 0.00937, 0.00938, 0.00939, or 0.00940
Sometimes it is. In some embodiments, the ratio of CD34 ⁺ cells to lymphocytes is about 0
.0025 to about 0.0035, for example, about 0.00250, 0.00251, 0.00252, 0.00253, 0.00254, 0.00255,
0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.00261, 0.00262, 0.00263, 0.00264
, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.00272, 0.0027
3, 0.00274, 0.00275, 0.00276, 0.00277, 0.00278, 0.00279, 0.00280, 0.00281, 0.002
82, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290, 0.00
291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.0
0300, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0.00307, 0.
00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0
.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325,
0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333, 0.00334,
0.00335, 0.00336, 0.00337, 0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343
The ratio of CD34 ⁺ cells to lymphocytes is , 0.00344, 0.00345, 0.00346, 0.00347, 0.00348, 0.00349, or 0.00350. In some embodiments, the ratio of CD34 ⁺ cells to lymphocytes is about 0.0031.

In a further aspect, the present invention relates to a pharmaceutical composition comprising a population of hematopoietic stem cells or their progeny isolated from a mammalian donor (e.g., a human donor), wherein a single cell in said population is The ratio of CD34 ⁺ cells to spheres is about 0.0071 to about 0.0174. In some embodiments, the ratio of CD34 ⁺ cells to monocytes is about 0.00710, 0.00711, 0.00712, 0.00713, 0
.00714, 0.00715, 0.00716, 0.00717, 0.00718, 0.00719, 0.00720, 0.00721, 0.00722,
0.00723, 0.00724, 0.00725, 0.00726, 0.00727, 0.00728, 0.00729, 0.00730, 0.00731,
0.00732, 0.00733, 0.00734, 0.00735, 0.00736, 0.00737, 0.00738, 0.00739, 0.00740
, 0.00741, 0.00742, 0.00743, 0.00744, 0.00745, 0.00746, 0.00747, 0.00748, 0.0074
9, 0.00750, 0.00751, 0.00752, 0.00753, 0.00754, 0.00755, 0.00756, 0.00757, 0.007
58, 0.00759, 0.00760, 0.00761, 0.00762, 0.00763, 0.00764, 0.00765, 0.00766, 0.00
767, 0.00768, 0.00769, 0.00770, 0.00771, 0.00772, 0.00773, 0.00774, 0.00775, 0.0
0776, 0.00777, 0.00778, 0.00779, 0.00780, 0.00781, 0.00782, 0.00783, 0.00784, 0.
00785, 0.00786, 0.00787, 0.00788, 0.00789, 0.00790, 0.00791, 0.00792, 0.00793, 0
.00794, 0.00795, 0.00796, 0.00797, 0.00798, 0.00799, 0.00800, 0.00801, 0.00802,
0.00803, 0.00804, 0.00805, 0.00806, 0.00807, 0.00808, 0.00809, 0.00810, 0.00811,
0.00812, 0.00813, 0.00814, 0.00815, 0.00816, 0.00817, 0.00818, 0.00819, 0.00820
, 0.00821, 0.00822, 0.00823, 0.00824, 0.00825, 0.00826, 0.00827, 0.00828, 0.0082
9, 0.00830, 0.00831, 0.00832, 0.00833, 0.00834, 0.00835, 0.00836, 0.00837, 0.008
38, 0.00839, 0.00840, 0.00841, 0.00842, 0.00843, 0.00844, 0.00845, 0.00846, 0.00
847, 0.00848, 0.00849, 0.00850, 0.00851, 0.00852, 0.00853, 0.00854, 0.00855, 0.0
0856, 0.00857, 0.00858, 0.00859, 0.00860, 0.00861, 0.00862, 0.00863, 0.00864, 0.
00865, 0.00866, 0.00867, 0.00868, 0.00869, 0.00870, 0.00871, 0.00872, 0.00873, 0
.00874, 0.00875, 0.00876, 0.00877, 0.00878, 0.00879, 0.00880, 0.00881, 0.00882,
0.00883, 0.00884, 0.00885, 0.00886, 0.00887, 0.00888, 0.00889, 0.00890, 0.00891,
0.00892, 0.00893, 0.00894, 0.00895, 0.00896, 0.00897, 0.00898, 0.00899, 0.00900
, 0.00901, 0.00902, 0.00903, 0.00904, 0.00905, 0.00906, 0.00907, 0.00908, 0.0090
9, 0.00910, 0.00911, 0.00912, 0.00913, 0.00914, 0.00915, 0.00916, 0.00917, 0.009
18, 0.00919, 0.00920, 0.00921, 0.00922, 0.00923, 0.00924, 0.00925, 0.00926, 0.00
927, 0.00928, 0.00929, 0.00930, 0.00931, 0.00932, 0.00933, 0.00934, 0.00935, 0.0
0936, 0.00937, 0.00938, 0.00939, 0.00940, 0.00941, 0.00942, 0.00943, 0.00944, 0.
00945, 0.00946, 0.00947, 0.00948, 0.00949, 0.00950, 0.00951, 0.00952, 0.00953, 0
.00954, 0.00955, 0.00956, 0.00957, 0.00958, 0.00959, 0.00960, 0.00961, 0.00962,
0.00963, 0.00964, 0.00965, 0.00966, 0.00967, 0.00968, 0.00969, 0.00970, 0.00971,
0.00972, 0.00973, 0.00974, 0.00975, 0.00976, 0.00977, 0.00978, 0.00979, 0.00980
, 0.00981, 0.00982, 0.00983, 0.00984, 0.00985, 0.00986, 0.00987, 0.00988, 0.0098
9, 0.00990, 0.00991, 0.00992, 0.00993, 0.00994, 0.00995, 0.00996, 0.00997, 0.009
98, 0.00999, 0.0100, 0.0101, 0.0103, 0.0104, 0.0105, 0.0106, 0.0107, 0.0108, 0.0
109, 0.0110, 0.0111, 0.0112, 0.0113, 0.0114, 0.0115, 0.0116, 0.0117, 0.0118, 0.0
119, 0.0120, 0.0121, 0.0122, 0.0123, 0.0124, 0.0125, 0.0126, 0.0127, 0.0128, 0.0
129, 0.0130, 0.0131, 0.0132, 0.0133, 0.0134, 0.0135, 0.0136, 0.0137, 0.0138, 0.0
139, 0.0140, 0.0141, 0.0142, 0.0143, 0.0144, 0.0145, 0.0146, 0.0147, 0.0148, 0.0
149, 0.0150, 0.0151, 0.0152, 0.0153, 0.0154, 0.0155, 0.0156, 0.0157, 0.0158, 0.0
159, 0.0160, 0.0161, 0.0162, 0.0163, 0.0164, 0.0165, 0.0166, 0.0167, 0.0168, 0.0
169, 0.0170, 0.0171, 0.0172, 0.0173, or 0.0174. In some embodiments, the ratio of CD34 ⁺ cells to monocytes is about 0.0100 to about 0.0140, e.g., about 0.0100,0
.0101, 0.0103, 0.0104, 0.0105, 0.0106, 0.0107, 0.0108, 0.0109, 0.0110, 0.0111, 0
.0112, 0.0113, 0.0114, 0.0115, 0.0116, 0.0117, 0.0118, 0.0119, 0.0120, 0.0121, 0
.0122, 0.0123, 0.0124, 0.0125, 0.0126, 0.0127, 0.0128, 0.0129, 0.0130, 0.0131, 0
The ratio of CD34 ⁺ cells to monocytes is .0132, 0.0133, 0.0134, 0.0135, 0.0136, 0.0137, 0.0138, 0.0139, or 0.0140. In some embodiments, CD34 ⁺ on lymphocytes
The cell ratio is approximately 0.0118.

In a further aspect, the invention relates to a pharmaceutical composition comprising a population of hematopoietic stem cells or their progeny isolated from a mammalian donor (e.g., a human donor), wherein CD34 in said population The frequency of ⁺ cells is about 0.051% to about 0.140%. In some embodiments, the population of cells is about 0.051%, 0.052%, 0.053%, 0.054%, 0.055%, 0.056%, 0.057%, 0.058%
, 0.059%, 0.060%, 0.061%, 0.062%, 0.063%, 0.064%, 0.065%, 0.066%, 0.067%, 0.068%
, 0.069%, 0.070%, 0.071%, 0.072%, 0.073%, 0.074%, 0.075%, 0.076%, 0.077%, 0.078%
, 0.079%, 0.080%, 0.081%, 0.082%, 0.083%, 0.084%, 0.085%, 0.086%, 0.087%, 0.088%
, 0.089%, 0.090%, 0.091%, 0.092%, 0.093%, 0.094%, 0.095%, 0.096%, 0.097%, 0.098%
, 0.099%, 0.100%, 0.101%, 0.102%, 0.103%, 0.104%, 0.105%, 0.106%, 0.107%, 0.108%
, 0.109%, 0.110%, 0.111%, 0.112%, 0.113%, 0.114%, 0.115%, 0.116%, 0.117%, 0.118%
, 0.119%, 0.120%, 0.121%, 0.122%, 0.123%, 0.124%, 0.125%, 0.126%, 0.127%, 0.128%
, 0.129%, 0.130%, 0.131%, 0.132%, 0.133%, 0.134%, 0.135%, 0.136%, 0.137%, 0.138%
The frequency of CD34 ⁺ cells may be , 0.139%, or 0.140%. In some embodiments,
The cell population has a frequency of CD34 ⁺ cells of about 0.080% to about 0.120%, e.g., about 0.080%, 0.081%, 0.0
82%, 0.083%, 0.084%, 0.085%, 0.086%, 0.087%, 0.088%, 0.089%, 0.090%, 0.091%, 0.0
92%, 0.093%, 0.094%, 0.095%, 0.096%, 0.097%, 0.098%, 0.099%, 0.100%, 0.101%, 0.1
02%, 0.103%, 0.104%, 0.105%, 0.106%, 0.107%, 0.108%, 0.109%, 0.110%, 0.111%, 0.1
12%, 0.113%, 0.114%, 0.115%, 0.116%, 0.117%, 0.118%, 0.119%, or 0.120% CD34 ⁺
is the frequency of cells. In some embodiments, the population of cells has a frequency of CD34 ⁺ cells of about 0.097%.

In a further aspect, the invention relates to a pharmaceutical composition comprising a population of hematopoietic stem cells or progeny thereof isolated from a mammalian donor (e.g., a human donor), wherein leukocytes in said population The ratio of CD34 ⁺ CD90 + CD45RA ⁻ cells to CD34 + CD90 ⁺ CD45RA − cells is about 0.0003 to about 0.0016. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to white blood cells is about 0.000
30, 0.00031, 0.00032, 0.00033, 0.00034, 0.00035, 0.00036, 0.00037, 0.00038, 0.00
039, 0.00040, 0.00041, 0.00042, 0.00043, 0.00044, 0.00045, 0.00046, 0.00047, 0.0
0048, 0.00049, 0.00050, 0.00051, 0.00052, 0.00053, 0.00054, 0.00055, 0.00056, 0.
00057, 0.00058, 0.00059, 0.00060, 0.00061, 0.00062, 0.00063, 0.00064, 0.00065, 0
.00066, 0.00067, 0.00068, 0.00069, 0.00070, 0.00071, 0.00072, 0.00073, 0.00074,
0.00075, 0.00076, 0.00077, 0.00078, 0.00079, 0.00080, 0.00081, 0.00082, 0.00083,
0.00084, 0.00085, 0.00086, 0.00087, 0.00088, 0.00089, 0.00090, 0.00091, 0.00092
, 0.00093, 0.00094, 0.00095, 0.00096, 0.00097, 0.00098, 0.00099, 0.00100, 0.0010
1, 0.00102, 0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108, 0.00109, 0.001
10, 0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117, 0.00118, 0.00
119, 0.00120, 0.00121, 0.00122, 0.00123, 0.00124, 0.00125, 0.00126, 0.00127, 0.0
0128, 0.00129, 0.00130, 0.00131, 0.00132, 0.00133, 0.00134, 0.00135, 0.00136, 0.
00137, 0.00138, 0.00139, 0.00140, 0.00141, 0.00142, 0.00143, 0.00144, 0.00145, 0
.00146, 0.00147, 0.00148, 0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154,
May be 0.00155, 0.00156, 0.00157, 0.00158, 0.00159, or 0.00160. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to white blood cells is about 0.0006 to about 0.
.0012, e.g. approximately 0.00060, 0.00061, 0.00062, 0.00063, 0.00064, 0.00065, 0.00066,
0.00067, 0.00068, 0.00069, 0.00070, 0.00071, 0.00072, 0.00073, 0.00074, 0.00075,
0.00076, 0.00077, 0.00078, 0.00079, 0.00080, 0.00081, 0.00082, 0.00083, 0.00084
, 0.00085, 0.00086, 0.00087, 0.00088, 0.00089, 0.00090, 0.00091, 0.00092, 0.0009
3, 0.00094, 0.00095, 0.00096, 0.00097, 0.00098, 0.00099, 0.00100, 0.00101, 0.001
02, 0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108, 0.00109, 0.00110, 0.00
The ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to white blood cells is 111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117, 0.00118, 0.00119, or 0.00120. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to white blood cells is about 0.0009.

In a further aspect, the invention relates to a pharmaceutical composition comprising a population of hematopoietic stem cells or their progeny isolated from a mammalian donor (e.g., a human donor), wherein a preferred population of hematopoietic stem cells or progeny thereof is The ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to mesospheres is about 0.0007 to about 0.0043. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to neutrophils is about 0.000
70, 0.00071, 0.00072, 0.00073, 0.00074, 0.00075, 0.00076, 0.00077, 0.00078, 0.00
079, 0.00080, 0.00081, 0.00082, 0.00083, 0.00084, 0.00085, 0.00086, 0.00087, 0.0
0088, 0.00089, 0.00090, 0.00091, 0.00092, 0.00093, 0.00094, 0.00095, 0.00096, 0.
00097, 0.00098, 0.00099, 0.00100, 0.00101, 0.00102, 0.00103, 0.00104, 0.00105, 0
.00106, 0.00107, 0.00108, 0.00109, 0.00110, 0.00111, 0.00112, 0.00113, 0.00114,
0.00115, 0.00116, 0.00117, 0.00118, 0.00119, 0.00120, 0.00121, 0.00122, 0.00123,
0.00124, 0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00132
, 0.00133, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.0014
1, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.001
50, 0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158, 0.00
159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164, 0.00165, 0.00166, 0.00167, 0.0
0168, 0.00169, 0.00170, 0.00171, 0.00172, 0.00173, 0.00174, 0.00175, 0.00176, 0.
00177, 0.00178, 0.00179, 0.00180, 0.00181, 0.00182, 0.00183, 0.00184, 0.00185, 0
.00186, 0.00187, 0.00188, 0.00189, 0.00190, 0.00191, 0.00192, 0.00193, 0.00194,
0.00195, 0.00196, 0.00197, 0.00198, 0.00199, 0.00200, 0.00201, 0.00202, 0.00203,
0.00204, 0.00205, 0.00206, 0.00207, 0.00208, 0.00209, 0.00210, 0.00211, 0.00212
, 0.00213, 0.00214, 0.00215, 0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.0022
1, 0.00222, 0.00223, 0.00224, 0.00225, 0.00226, 0.00227, 0.00228, 0.00229, 0.002
30, 0.00231, 0.00232, 0.00233, 0.00234, 0.00235, 0.00236, 0.00237, 0.00238, 0.00
239, 0.00240, 0.00241, 0.00242, 0.00243, 0.00244, 0.00245, 0.00246, 0.00247, 0.0
0248, 0.00249, 0.00250, 0.00251, 0.00252, 0.00253, 0.00254, 0.00255, 0.00256, 0.
00257, 0.00258, 0.00259, 0.00260, 0.00261, 0.00262, 0.00263, 0.00264, 0.00265, 0
.00266, 0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.00272, 0.00273, 0.00274,
0.00275, 0.00276, 0.00277, 0.00278, 0.00279, 0.00280, 0.00281, 0.00282, 0.00283,
0.00284, 0.00285, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290, 0.00291, 0.00292
, 0.00293, 0.00294, 0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.0030
0, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308, 0.003
09, 0.00310, 0.00311, 0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0.00317, 0.00
318, 0.00319, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325, 0.00326, 0.0
0327, 0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00335, 0.
00336, 0.00337, 0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343, 0.00344, 0
.00345, 0.00346, 0.00347, 0.00348, 0.00349, 0.00350, 0.00351, 0.00352, 0.00353,
0.00354, 0.00355, 0.00356, 0.00357, 0.00358, 0.00359, 0.00360, 0.00361, 0.00362,
0.00363, 0.00364, 0.00365, 0.00366, 0.00367, 0.00368, 0.00369, 0.00370, 0.00371
, 0.00372, 0.00373, 0.00374, 0.00375, 0.00376, 0.00377, 0.00378, 0.00379, 0.0038
0, 0.00381, 0.00382, 0.00383, 0.00384, 0.00385, 0.00386, 0.00387, 0.00388, 0.003
89, 0.00390, 0.00391, 0.00392, 0.00393, 0.00394, 0.00395, 0.00396, 0.00397, 0.00
398, 0.00399, 0.00400, 0.00401, 0.00402, 0.00403, 0.00404, 0.00405, 0.00406, 0.0
0407, 0.00408, 0.00409, 0.00410, 0.00411, 0.00412, 0.00413, 0.00414, 0.00415, 0.
00416, 0.00417, 0.00418, 0.00419, 0.00420, 0.00421, 0.00422, 0.00423, 0.00424, 0
May be .00425, 0.00426, 0.00427, 0.00428, 0.00429, or 0.00430. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to neutrophils is about 0.0014 to about 0.0.
034, for example, approximately 0.00140, 0.00141, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.
00147, 0.00148, 0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0
.00156, 0.00157, 0.00158, 0.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164,
0.00165, 0.00166, 0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172, 0.00173,
0.00174, 0.00175, 0.00176, 0.00177, 0.00178, 0.00179, 0.00180, 0.00181, 0.00182
, 0.00183, 0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189, 0.00190, 0.0019
1, 0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197, 0.00198, 0.00199, 0.002
00, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205, 0.00206, 0.00207, 0.00208, 0.00
209, 0.00210, 0.00211, 0.00212, 0.00213, 0.00214, 0.00215, 0.00216, 0.00217, 0.0
0218, 0.00219, 0.00220, 0.00221, 0.00222, 0.00223, 0.00224, 0.00225, 0.00226, 0.
00227, 0.00228, 0.00229, 0.00230, 0.00231, 0.00232, 0.00233, 0.00234, 0.00235, 0
.00236, 0.00237, 0.00238, 0.00239, 0.00240, 0.00241, 0.00242, 0.00243, 0.00244,
0.00245, 0.00246, 0.00247, 0.00248, 0.00249, 0.00250, 0.00251, 0.00252, 0.00253,
0.00254, 0.00255, 0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.00261, 0.00262
, 0.00263, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270, 0.0027
1, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00277, 0.00278, 0.00279, 0.002
80, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288, 0.00
289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.00297, 0.0
0298, 0.00299, 0.00300, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305, 0.
00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313, 0.00314, 0
.00315, 0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0.00323,
0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331, 0.00332,
The ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to neutrophils is 0.00333, 0.00334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339, or 0.00340. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to neutrophils is about 0.0024.

In another aspect, the present invention relates to a pharmaceutical composition comprising a population of hematopoietic stem cells or their progeny isolated from a mammalian donor (e.g., a human donor), wherein lymphocytes in said population The ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to spheres is about 0.0008 to about 0.0069. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to lymphocytes is about 0.0
0080, 0.00081, 0.00082, 0.00083, 0.00084, 0.00085, 0.00086, 0.00087, 0.00088, 0.
00089, 0.00090, 0.00091, 0.00092, 0.00093, 0.00094, 0.00095, 0.00096, 0.00097, 0
.00098, 0.00099, 0.00100, 0.00101, 0.00102, 0.00103, 0.00104, 0.00105, 0.00106,
0.00107, 0.00108, 0.00109, 0.00110, 0.00111, 0.00112, 0.00113, 0.00114, 0.00115,
0.00116, 0.00117, 0.00118, 0.00119, 0.00120, 0.00121, 0.00122, 0.00123, 0.00124
, 0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00132, 0.0013
3, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.00141, 0.001
42, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.00150, 0.00
151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158, 0.00159, 0.0
0160, 0.00161, 0.00162, 0.00163, 0.00164, 0.00165, 0.00166, 0.00167, 0.00168, 0.
00169, 0.00170, 0.00171, 0.00172, 0.00173, 0.00174, 0.00175, 0.00176, 0.00178, 0
.00179, 0.00180, 0.00181, 0.00182, 0.00183, 0.00184, 0.00185, 0.00186, 0.00187,
0.00188, 0.00189, 0.00190, 0.00191, 0.00192, 0.00193, 0.00194, 0.00195, 0.00196,
0.00197, 0.00198, 0.00199, 0.00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205
, 0.00206, 0.00207, 0.00208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00213, 0.0021
4, 0.00215, 0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.00222, 0.002
23, 0.00224, 0.00225, 0.00226, 0.00227, 0.00228, 0.00229, 0.00230, 0.00231, 0.00
232, 0.00233, 0.00234, 0.00235, 0.00236, 0.00237, 0.00238, 0.00239, 0.00240, 0.0
0241, 0.00242, 0.00243, 0.00244, 0.00245, 0.00246, 0.00247, 0.00248, 0.00249, 0.
00250, 0.00251, 0.00252, 0.00253, 0.00254, 0.00255, 0.00256, 0.00257, 0.00258, 0
.00259, 0.00260, 0.00261, 0.00262, 0.00263, 0.00264, 0.00265, 0.00266, 0.00267,
0.00268, 0.00269, 0.00270, 0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276,
0.00278, 0.00279, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286
, 0.00287, 0.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.0029
5, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.00301, 0.00302, 0.00303, 0.003
04, 0.00305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00
313, 0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.0
0322, 0.00323, 0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.
00331, 0.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339, 0
.00340, 0.00341, 0.00342, 0.00343, 0.00344, 0.00345, 0.00346, 0.00347, 0.00348,
0.00349, 0.00350, 0.00351, 0.00352, 0.00353, 0.00354, 0.00355, 0.00356, 0.00357,
0.00358, 0.00359, 0.00360, 0.00361, 0.00362, 0.00363, 0.00364, 0.00365, 0.00366
, 0.00367, 0.00368, 0.00369, 0.00370, 0.00371, 0.00372, 0.00373, 0.00374, 0.0037
5, 0.00376, 0.00378, 0.00379, 0.00380, 0.00381, 0.00382, 0.00383, 0.00384, 0.003
85, 0.00386, 0.00387, 0.00388, 0.00389, 0.00390, 0.00391, 0.00392, 0.00393, 0.00
394, 0.00395, 0.00396, 0.00397, 0.00398, 0.00399, 0.00401, 0.00402, 0.00403, 0.0
0404, 0.00405, 0.00406, 0.00407, 0.00408, 0.00409, 0.00410, 0.00411, 0.00412, 0.
00413, 0.00414, 0.00415, 0.00416, 0.00417, 0.00418, 0.00419, 0.00420, 0.00421, 0
.00422, 0.00423, 0.00424, 0.00425, 0.00426, 0.00427, 0.00428, 0.00429, 0.00430,
0.00431, 0.00432, 0.00433, 0.00434, 0.00435, 0.00436, 0.00437, 0.00438, 0.00439,
0.00440, 0.00441, 0.00442, 0.00443, 0.00444, 0.00445, 0.00446, 0.00447, 0.00448
, 0.00449, 0.00450, 0.00451, 0.00452, 0.00453, 0.00454, 0.00455, 0.00456, 0.0045
7, 0.00458, 0.00459, 0.00460, 0.00461, 0.00462, 0.00463, 0.00464, 0.00465, 0.004
66, 0.00467, 0.00468, 0.00469, 0.00470, 0.00471, 0.00472, 0.00473, 0.00474, 0.00
475, 0.00476, 0.00478, 0.00479, 0.00480, 0.00481, 0.00482, 0.00483, 0.00484, 0.0
0485, 0.00486, 0.00487, 0.00488, 0.00489, 0.00490, 0.00491, 0.00492, 0.00493, 0.
00494, 0.00495, 0.00496, 0.00497, 0.00498, 0.00499, 0.00500, 0.00501, 0.00502, 0
.00503, 0.00504, 0.00505, 0.00506, 0.00507, 0.00508, 0.00509, 0.00510, 0.00511,
0.00512, 0.00513, 0.00514, 0.00515, 0.00516, 0.00517, 0.00518, 0.00519, 0.00520,
0.00521, 0.00522, 0.00523, 0.00524, 0.00525, 0.00526, 0.00527, 0.00528, 0.00529
, 0.00530, 0.00531, 0.00532, 0.00533, 0.00534, 0.00535, 0.00536, 0.00537, 0.0053
8, 0.00539, 0.00540, 0.00541, 0.00542, 0.00543, 0.00544, 0.00545, 0.00546, 0.005
47, 0.00548, 0.00549, 0.00550, 0.00551, 0.00552, 0.00553, 0.00554, 0.00555, 0.00
556, 0.00557, 0.00558, 0.00559, 0.00560, 0.00561, 0.00562, 0.00563, 0.00564, 0.0
0565, 0.00566, 0.00567, 0.00568, 0.00569, 0.00570, 0.00571, 0.00572, 0.00573, 0.
00574, 0.00575, 0.00576, 0.00578, 0.00579, 0.00580, 0.00581, 0.00582, 0.00583, 0
.00584, 0.00585, 0.00586, 0.00587, 0.00588, 0.00589, 0.00590, 0.00591, 0.00592,
0.00593, 0.00594, 0.00595, 0.00596, 0.00597, 0.00598, 0.00599, 0.00600, 0.00601,
0.00602, 0.00603, 0.00604, 0.00605, 0.00606, 0.00607, 0.00608, 0.00609, 0.00610
, 0.00611, 0.00612, 0.00613, 0.00614, 0.00615, 0.00616, 0.00617, 0.00618, 0.0061
9, 0.00620, 0.00621, 0.00622, 0.00623, 0.00624, 0.00625, 0.00626, 0.00627, 0.006
28, 0.00629, 0.00630, 0.00631, 0.00632, 0.00633, 0.00634, 0.00635, 0.00636, 0.00
637, 0.00638, 0.00639, 0.00640, 0.00641, 0.00642, 0.00643, 0.00644, 0.00645, 0.0
0646, 0.00647, 0.00648, 0.00649, 0.00650, 0.00651, 0.00652, 0.00653, 0.00654, 0.
00655, 0.00656, 0.00657, 0.00658, 0.00659, 0.00660, 0.00661, 0.00662, 0.00663, 0
.00664, 0.00665, 0.00666, 0.00667, 0.00668, 0.00669, 0.00670, 0.00671, 0.00672,
0.00673, 0.00674, 0.00675, 0.00676, 0.00678, 0.00679, 0.00680, 0.00681, 0.00682,
May be 0.00683, 0.00684, 0.00685, 0.00686, 0.00687, 0.00688, 0.00689, or 0.00690. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to lymphocytes is about 0.0011 to about 0.0031, e.g., about 0.00110, 0.00111, 0.00112, 0.00113, 0.0011
4, 0.00115, 0.00116, 0.00117, 0.00118, 0.00119, 0.00120, 0.00121, 0.00122, 0.001
23, 0.00124, 0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00
132, 0.00133, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.0
0141, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.
00150, 0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158, 0
.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164, 0.00165, 0.00166, 0.00167,
0.00168, 0.00169, 0.00170, 0.00171, 0.00172, 0.00173, 0.00174, 0.00175, 0.00176,
0.00178, 0.00179, 0.00180, 0.00181, 0.00182, 0.00183, 0.00184, 0.00185, 0.00186
, 0.00187, 0.00188, 0.00189, 0.00190, 0.00191, 0.00192, 0.00193, 0.00194, 0.0019
5, 0.00196, 0.00197, 0.00198, 0.00199, 0.00200, 0.00201, 0.00202, 0.00203, 0.002
04, 0.00205, 0.00206, 0.00207, 0.00208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00
213, 0.00214, 0.00215, 0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.0
0222, 0.00223, 0.00224, 0.00225, 0.00226, 0.00227, 0.00228, 0.00229, 0.00230, 0.
00231, 0.00232, 0.00233, 0.00234, 0.00235, 0.00236, 0.00237, 0.00238, 0.00239, 0
.00240, 0.00241, 0.00242, 0.00243, 0.00244, 0.00245, 0.00246, 0.00247, 0.00248,
0.00249, 0.00250, 0.00251, 0.00252, 0.00253, 0.00254, 0.00255, 0.00256, 0.00257,
0.00258, 0.00259, 0.00260, 0.00261, 0.00262, 0.00263, 0.00264, 0.00265, 0.00266
, 0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.00272, 0.00273, 0.00274, 0.0027
5, 0.00276, 0.00278, 0.00279, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.002
85, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00
294, 0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.00301, 0.00302, 0.0
The ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to lymphocytes is 0.0303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308, 0.00309, or 0.00310. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to lymphocytes is about 0.0021.

In a further aspect, the present invention relates to a pharmaceutical composition comprising a population of hematopoietic stem cells or their progeny isolated from a mammalian donor (e.g., a human donor), wherein the cells in said population are The ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to spheres is about 0.0028 to about 0.0130.
In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to monocytes is about 0.00280,
0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288, 0.00289,
0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.00297, 0.00298
, 0.00299, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0.0030
7, 0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313, 0.00314, 0.00315, 0.003
16, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00
325, 0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333, 0.0
0334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.
00343, 0.00344, 0.00345, 0.00346, 0.00347, 0.00348, 0.00349, 0.00350, 0.00351, 0
.00352, 0.00353, 0.00354, 0.00355, 0.00356, 0.00357, 0.00358, 0.00359, 0.00360,
0.00361, 0.00362, 0.00363, 0.00364, 0.00365, 0.00366, 0.00367, 0.00368, 0.00369,
0.00370, 0.00371, 0.00372, 0.00373, 0.00374, 0.00375, 0.00376, 0.00378, 0.00379
, 0.00380, 0.00381, 0.00382, 0.00383, 0.00384, 0.00385, 0.00386, 0.00387, 0.0038
8, 0.00389, 0.00390, 0.00391, 0.00392, 0.00393, 0.00394, 0.00395, 0.00396, 0.003
97, 0.00398, 0.00399, 0.00401, 0.00402, 0.00403, 0.00404, 0.00405, 0.00406, 0.00
407, 0.00408, 0.00409, 0.00410, 0.00411, 0.00412, 0.00413, 0.00414, 0.00415, 0.0
0416, 0.00417, 0.00418, 0.00419, 0.00420, 0.00421, 0.00422, 0.00423, 0.00424, 0.
00425, 0.00426, 0.00427, 0.00428, 0.00429, 0.00430, 0.00431, 0.00432, 0.00433, 0
.00434, 0.00435, 0.00436, 0.00437, 0.00438, 0.00439, 0.00440, 0.00441, 0.00442,
0.00443, 0.00444, 0.00445, 0.00446, 0.00447, 0.00448, 0.00449, 0.00450, 0.00451,
0.00452, 0.00453, 0.00454, 0.00455, 0.00456, 0.00457, 0.00458, 0.00459, 0.00460
, 0.00461, 0.00462, 0.00463, 0.00464, 0.00465, 0.00466, 0.00467, 0.00468, 0.0046
9, 0.00470, 0.00471, 0.00472, 0.00473, 0.00474, 0.00475, 0.00476, 0.00478, 0.004
79, 0.00480, 0.00481, 0.00482, 0.00483, 0.00484, 0.00485, 0.00486, 0.00487, 0.00
488, 0.00489, 0.00490, 0.00491, 0.00492, 0.00493, 0.00494, 0.00495, 0.00496, 0.0
0497, 0.00498, 0.00499, 0.00500, 0.00501, 0.00502, 0.00503, 0.00504, 0.00505, 0.
00506, 0.00507, 0.00508, 0.00509, 0.00510, 0.00511, 0.00512, 0.00513, 0.00514, 0
.00515, 0.00516, 0.00517, 0.00518, 0.00519, 0.00520, 0.00521, 0.00522, 0.00523,
0.00524, 0.00525, 0.00526, 0.00527, 0.00528, 0.00529, 0.00530, 0.00531, 0.00532,
0.00533, 0.00534, 0.00535, 0.00536, 0.00537, 0.00538, 0.00539, 0.00540, 0.00541
, 0.00542, 0.00543, 0.00544, 0.00545, 0.00546, 0.00547, 0.00548, 0.00549, 0.0055
0, 0.00551, 0.00552, 0.00553, 0.00554, 0.00555, 0.00556, 0.00557, 0.00558, 0.005
59, 0.00560, 0.00561, 0.00562, 0.00563, 0.00564, 0.00565, 0.00566, 0.00567, 0.00
568, 0.00569, 0.00570, 0.00571, 0.00572, 0.00573, 0.00574, 0.00575, 0.00576, 0.0
0578, 0.00579, 0.00580, 0.00581, 0.00582, 0.00583, 0.00584, 0.00585, 0.00586, 0.
00587, 0.00588, 0.00589, 0.00590, 0.00591, 0.00592, 0.00593, 0.00594, 0.00595, 0
.00596, 0.00597, 0.00598, 0.00599, 0.00600, 0.00601, 0.00602, 0.00603, 0.00604,
0.00605, 0.00606, 0.00607, 0.00608, 0.00609, 0.00610, 0.00611, 0.00612, 0.00613,
0.00614, 0.00615, 0.00616, 0.00617, 0.00618, 0.00619, 0.00620, 0.00621, 0.00622
, 0.00623, 0.00624, 0.00625, 0.00626, 0.00627, 0.00628, 0.00629, 0.00630, 0.0063
1, 0.00632, 0.00633, 0.00634, 0.00635, 0.00636, 0.00637, 0.00638, 0.00639, 0.006
40, 0.00641, 0.00642, 0.00643, 0.00644, 0.00645, 0.00646, 0.00647, 0.00648, 0.00
649, 0.00650, 0.00651, 0.00652, 0.00653, 0.00654, 0.00655, 0.00656, 0.00657, 0.0
0658, 0.00659, 0.00660, 0.00661, 0.00662, 0.00663, 0.00664, 0.00665, 0.00666, 0.
00667, 0.00668, 0.00669, 0.00670, 0.00671, 0.00672, 0.00673, 0.00674, 0.00675, 0
.00676, 0.00678, 0.00679, 0.00680, 0.00681, 0.00682, 0.00683, 0.00684, 0.00685,
0.00686, 0.00687, 0.00688, 0.00689, 0.00690, 0.00691, 0.00692, 0.00693, 0.00694,
0.00695, 0.00696, 0.00697, 0.00698, 0.00699, 0.00700, 0.00701, 0.00702, 0.00703
, 0.00704, 0.00705, 0.00706, 0.00707, 0.00708, 0.00709, 0.00710, 0.00711, 0.0071
2, 0.00713, 0.00714, 0.00715, 0.00716, 0.00717, 0.00718, 0.00719, 0.00720, 0.007
21, 0.00722, 0.00723, 0.00724, 0.00725, 0.00726, 0.00727, 0.00728, 0.00729, 0.00
730, 0.00731, 0.00732, 0.00733, 0.00734, 0.00735, 0.00736, 0.00737, 0.00738, 0.0
0739, 0.00740, 0.00741, 0.00742, 0.00743, 0.00744, 0.00745, 0.00746, 0.00747, 0.
00748, 0.00749, 0.00750, 0.00751, 0.00752, 0.00753, 0.00754, 0.00755, 0.00756, 0
.00757, 0.00758, 0.00759, 0.00760, 0.00761, 0.00762, 0.00763, 0.00764, 0.00765,
0.00766, 0.00767, 0.00768, 0.00769, 0.00770, 0.00771, 0.00772, 0.00773, 0.00774,
0.00775, 0.00776, 0.00777, 0.00778, 0.00779, 0.00780, 0.00781, 0.00782, 0.00783
, 0.00784, 0.00785, 0.00786, 0.00787, 0.00788, 0.00789, 0.00790, 0.00791, 0.0079
2, 0.00793, 0.00794, 0.00795, 0.00796, 0.00797, 0.00798, 0.00799, 0.00800, 0.008
01, 0.00802, 0.00803, 0.00804, 0.00805, 0.00806, 0.00807, 0.00808, 0.00809, 0.00
810, 0.00811, 0.00812, 0.00813, 0.00814, 0.00815, 0.00816, 0.00817, 0.00818, 0.0
0819, 0.00820, 0.00821, 0.00822, 0.00823, 0.00824, 0.00825, 0.00826, 0.00827, 0.
00828, 0.00829, 0.00830, 0.00831, 0.00832, 0.00833, 0.00834, 0.00835, 0.00836, 0
.00837, 0.00838, 0.00839, 0.00840, 0.00841, 0.00842, 0.00843, 0.00844, 0.00845,
0.00846, 0.00847, 0.00848, 0.00849, 0.00850, 0.00851, 0.00852, 0.00853, 0.00854,
0.00855, 0.00856, 0.00857, 0.00858, 0.00859, 0.00860, 0.00861, 0.00862, 0.00863
, 0.00864, 0.00865, 0.00866, 0.00867, 0.00868, 0.00869, 0.00870, 0.00871, 0.0087
2, 0.00873, 0.00874, 0.00875, 0.00876, 0.00877, 0.00878, 0.00879, 0.00880, 0.008
81, 0.00882, 0.00883, 0.00884, 0.00885, 0.00886, 0.00887, 0.00888, 0.00889, 0.00
890, 0.00891, 0.00892, 0.00893, 0.00894, 0.00895, 0.00896, 0.00897, 0.00898, 0.0
0899, 0.00900, 0.00901, 0.00902, 0.00903, 0.00904, 0.00905, 0.00906, 0.00907, 0.
00908, 0.00909, 0.00910, 0.00911, 0.00912, 0.00913, 0.00914, 0.00915, 0.00916, 0
.00917, 0.00918, 0.00919, 0.00920, 0.00921, 0.00922, 0.00923, 0.00924, 0.00925,
0.00926, 0.00927, 0.00928, 0.00929, 0.00930, 0.00931, 0.00932, 0.00933, 0.00934,
0.00935, 0.00936, 0.00937, 0.00938, 0.00939, 0.00940, 0.00941, 0.00942, 0.00943
, 0.00944, 0.00945, 0.00946, 0.00947, 0.00948, 0.00949, 0.00950, 0.00951, 0.0095
2, 0.00953, 0.00954, 0.00955, 0.00956, 0.00957, 0.00958, 0.00959, 0.00960, 0.009
61, 0.00962, 0.00963, 0.00964, 0.00965, 0.00966, 0.00967, 0.00968, 0.00969, 0.00
970, 0.00971, 0.00972, 0.00973, 0.00974, 0.00975, 0.00976, 0.00977, 0.00978, 0.0
0979, 0.00980, 0.00981, 0.00982, 0.00983, 0.00984, 0.00985, 0.00986, 0.00987, 0.
00988, 0.00989, 0.00990, 0.00991, 0.00992, 0.00993, 0.00994, 0.00995, 0.00996, 0
.00997, 0.00998, 0.00999, 0.0100, 0.0101, 0.0103, 0.0104, 0.0105, 0.0106, 0.0107
, 0.0108, 0.0109, 0.0110, 0.0111, 0.0112, 0.0113, 0.0114, 0.0115, 0.0116, 0.0117
, 0.0118, 0.0119, 0.0120, 0.0121, 0.0122, 0.0123, 0.0124, 0.0125, 0.0126, 0.0127
, 0.0128, 0.0129, or 0.0130. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to monocytes is about 0.0063 to about 0.0083, e.g., about 0.00630, 0.00
631, 0.00632, 0.00633, 0.00634, 0.00635, 0.00636, 0.00637, 0.00638, 0.00639, 0.0
0640, 0.00641, 0.00642, 0.00643, 0.00644, 0.00645, 0.00646, 0.00647, 0.00648, 0.
00649, 0.00650, 0.00651, 0.00652, 0.00653, 0.00654, 0.00655, 0.00656, 0.00657, 0
.00658, 0.00659, 0.00660, 0.00661, 0.00662, 0.00663, 0.00664, 0.00665, 0.00666,
0.00667, 0.00668, 0.00669, 0.00670, 0.00671, 0.00672, 0.00673, 0.00674, 0.00675,
0.00676, 0.00678, 0.00679, 0.00680, 0.00681, 0.00682, 0.00683, 0.00684, 0.00685
, 0.00686, 0.00687, 0.00688, 0.00689, 0.00690, 0.00691, 0.00692, 0.00693, 0.0069
4, 0.00695, 0.00696, 0.00697, 0.00698, 0.00699, 0.00700, 0.00701, 0.00702, 0.007
03, 0.00704, 0.00705, 0.00706, 0.00707, 0.00708, 0.00709, 0.00710, 0.00711, 0.00
712, 0.00713, 0.00714, 0.00715, 0.00716, 0.00717, 0.00718, 0.00719, 0.00720, 0.0
0721, 0.00722, 0.00723, 0.00724, 0.00725, 0.00726, 0.00727, 0.00728, 0.00729, 0.
00730, 0.00731, 0.00732, 0.00733, 0.00734, 0.00735, 0.00736, 0.00737, 0.00738, 0
.00739, 0.00740, 0.00741, 0.00742, 0.00743, 0.00744, 0.00745, 0.00746, 0.00747,
0.00748, 0.00749, 0.00750, 0.00751, 0.00752, 0.00753, 0.00754, 0.00755, 0.00756,
0.00757, 0.00758, 0.00759, 0.00760, 0.00761, 0.00762, 0.00763, 0.00764, 0.00765
, 0.00766, 0.00767, 0.00768, 0.00769, 0.00770, 0.00771, 0.00772, 0.00773, 0.0077
4, 0.00775, 0.00776, 0.00777, 0.00778, 0.00779, 0.00780, 0.00781, 0.00782, 0.007
83, 0.00784, 0.00785, 0.00786, 0.00787, 0.00788, 0.00789, 0.00790, 0.00791, 0.00
792, 0.00793, 0.00794, 0.00795, 0.00796, 0.00797, 0.00798, 0.00799, 0.00800, 0.0
0801, 0.00802, 0.00803, 0.00804, 0.00805, 0.00806, 0.00807, 0.00808, 0.00809, 0.
00810, 0.00811, 0.00812, 0.00813, 0.00814, 0.00815, 0.00816, 0.00817, 0.00818, 0
.00819, 0.00820, 0.00821, 0.00822, 0.00823, 0.00824, 0.00825, 0.00826, 0.00827,
The ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to monocytes is 0.00828, 0.00829, or 0.00830. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to monocytes is about 0.0073
It is.

In a further aspect, the invention relates to a pharmaceutical composition comprising a population of hematopoietic stem cells or progeny thereof isolated from a mammalian donor (e.g., a human donor), wherein CD34 in said population The ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to ⁺ cells is about 0.393 to about 0.745. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to CD34 ⁺ cells is about 0.
393, 0.394, 0.395, 0.396, 0.397, 0.398, 0.399, 0.401, 0.402, 0.403, 0.404, 0.405
, 0.406, 0.407, 0.408, 0.409, 0.410, 0.411, 0.412, 0.413, 0.414, 0.415, 0.416, 0
.417, 0.418, 0.419, 0.420, 0.421, 0.422, 0.423, 0.424, 0.425, 0.426, 0.427, 0.42
8, 0.429, 0.430, 0.431, 0.432, 0.433, 0.434, 0.435, 0.436, 0.437, 0.438, 0.439,
0.440, 0.441, 0.442, 0.443, 0.444, 0.445, 0.446, 0.447, 0.448, 0.449, 0.450, 0.4
51, 0.452, 0.453, 0.454, 0.455, 0.456, 0.457, 0.458, 0.459, 0.460, 0.461, 0.462,
0.463, 0.464, 0.465, 0.466, 0.467, 0.468, 0.469, 0.470, 0.471, 0.472, 0.473, 0.
474, 0.475, 0.476, 0.478, 0.479, 0.480, 0.481, 0.482, 0.483, 0.484, 0.485, 0.486
, 0.487, 0.488, 0.489, 0.490, 0.491, 0.492, 0.493, 0.494, 0.495, 0.496, 0.497, 0
.498, 0.499, 0.500, 0.501, 0.502, 0.503, 0.504, 0.505, 0.506, 0.507, 0.508, 0.50
9, 0.510, 0.511, 0.512, 0.513, 0.514, 0.515, 0.516, 0.517, 0.518, 0.519, 0.520,
0.521, 0.522, 0.523, 0.524, 0.525, 0.526, 0.527, 0.528, 0.529, 0.530, 0.531, 0.5
32, 0.533, 0.534, 0.535, 0.536, 0.537, 0.538, 0.539, 0.540, 0.541, 0.542, 0.543,
0.544, 0.545, 0.546, 0.547, 0.548, 0.549, 0.550, 0.551, 0.552, 0.553, 0.554, 0.
555, 0.556, 0.557, 0.558, 0.559, 0.560, 0.561, 0.562, 0.563, 0.564, 0.565, 0.566
, 0.567, 0.568, 0.569, 0.570, 0.571, 0.572, 0.573, 0.574, 0.575, 0.576, 0.578, 0
.579, 0.580, 0.581, 0.582, 0.583, 0.584, 0.585, 0.586, 0.587, 0.588, 0.589, 0.59
0, 0.591, 0.592, 0.593, 0.594, 0.595, 0.596, 0.597, 0.598, 0.599, 0.600, 0.601,
0.602, 0.603, 0.604, 0.605, 0.606, 0.607, 0.608, 0.609, 0.610, 0.611, 0.612, 0.6
13, 0.614, 0.615, 0.616, 0.617, 0.618, 0.619, 0.620, 0.621, 0.622, 0.623, 0.624,
0.625, 0.626, 0.627, 0.628, 0.629, 0.630, 0.631, 0.632, 0.633, 0.634, 0.635, 0.
636, 0.637, 0.638, 0.639, 0.640, 0.641, 0.642, 0.643, 0.644, 0.645, 0.646, 0.647
, 0.648, 0.649, 0.650, 0.651, 0.652, 0.653, 0.654, 0.655, 0.656, 0.657, 0.658, 0
.659, 0.660, 0.661, 0.662, 0.663, 0.664, 0.665, 0.666, 0.667, 0.668, 0.669, 0.67
0, 0.671, 0.672, 0.673, 0.674, 0.675, 0.676, 0.678, 0.679, 0.680, 0.681, 0.682,
0.683, 0.684, 0.685, 0.686, 0.687, 0.688, 0.689, 0.690, 0.691, 0.692, 0.693, 0.6
94, 0.695, 0.696, 0.697, 0.698, 0.699, 0.700, 0.701, 0.702, 0.703, 0.704, 0.705,
0.706, 0.707, 0.708, 0.709, 0.710, 0.711, 0.712, 0.713, 0.714, 0.715, 0.716, 0.
717, 0.718, 0.719, 0.720, 0.721, 0.722, 0.723, 0.724, 0.725, 0.726, 0.727, 0.728
, 0.729, 0.730, 0.731, 0.732, 0.733, 0.734, 0.735, 0.736, 0.737, 0.738, 0.739, 0
May be .740, 0.741, 0.742, 0.743, 0.744, or 0.745. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to CD34 ⁺ cells is about 0.625 to about 0.725, e.g., about 0.625, 0.626, 0.627, 0.628, 0.629, 0.630, 0.631, 0.632, 0.633, 0.634. , 0.635,
0.636, 0.637, 0.638, 0.639, 0.640, 0.641, 0.642, 0.643, 0.644, 0.645, 0.646, 0.
647, 0.648, 0.649, 0.650, 0.651, 0.652, 0.653, 0.654, 0.655, 0.656, 0.657, 0.658
, 0.659, 0.660, 0.661, 0.662, 0.663, 0.664, 0.665, 0.666, 0.667, 0.668, 0.669, 0
.670, 0.671, 0.672, 0.673, 0.674, 0.675, 0.676, 0.678, 0.679, 0.680, 0.681, 0.68
2, 0.683, 0.684, 0.685, 0.686, 0.687, 0.688, 0.689, 0.690, 0.691, 0.692, 0.693,
0.694, 0.695, 0.696, 0.697, 0.698, 0.699, 0.700, 0.701, 0.702, 0.703, 0.704, 0.7
05, 0.706, 0.707, 0.708, 0.709, 0.710, 0.711, 0.712, 0.713, 0.714, 0.715, 0.716,
The ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to CD34 ⁺ cells is 0.717, 0.718, 0.719, 0.720, 0.721, 0.722, 0.723, 0.724, or 0.725. In some embodiments, the ratio of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells to CD34 ⁺ cells is about 0.676.

In a further aspect, the invention relates to a pharmaceutical composition comprising a population of hematopoietic stem cells or progeny thereof isolated from a mammalian donor (e.g., a human donor), wherein CD34 in said population The frequency of ⁺ CD90 ⁺ CD45RA ⁻ cells is about 0.020% to about 0.110%. In some embodiments, the population of cells is about 0.020%, 0.021%, 0.022%, 0.023%, 0.024%, 0.025%,
0.026%, 0.027%, 0.028%, 0.029%, 0.030%, 0.031%, 0.032%, 0.033%, 0.034%, 0.035%,
0.036%, 0.037%, 0.038%, 0.039%, 0.040%, 0.041%, 0.042%, 0.043%, 0.044%, 0.045%,
0.046%, 0.047%, 0.048%, 0.049%, 0.050%, 0.051%, 0.052%, 0.053%, 0.054%, 0.055%,
0.056%, 0.057%, 0.058%, 0.059%, 0.060%, 0.061%, 0.062%, 0.063%, 0.064%, 0.065%,
0.066%, 0.067%, 0.068%, 0.069%, 0.070%, 0.071%, 0.072%, 0.073%, 0.074%, 0.075%,
0.076%, 0.077%, 0.078%, 0.079%, 0.080%, 0.081%, 0.082%, 0.083%, 0.084%, 0.085%,
0.086%, 0.087%, 0.088%, 0.089%, 0.090%, 0.091%, 0.092%, 0.093%, 0.094%, 0.095%,
0.096%, 0.097%, 0.098%, 0.099%, 0.100%, 0.101%, 0.102%, 0.103%, 0.104%, 0.105%,
The frequency of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells may be 0.106%, 0.107%, 0.108%, 0.109%, or 0.110%. In some embodiments, the cell population has about 0.046% to about 0.086% CD34 ⁺ CD9
Frequency of 0 ⁺ CD45RA ^- cells, e.g. approximately 0.046%, 0.047%, 0.048%, 0.049%, 0.050%, 0.051%, 0
.052%, 0.053%, 0.054%, 0.055%, 0.056%, 0.057%, 0.058%, 0.059%, 0.060%, 0.061%, 0
.062%, 0.063%, 0.064%, 0.065%, 0.066%, 0.067%, 0.068%, 0.069%, 0.070%, 0.071%, 0
.072%, 0.073%, 0.074%, 0.075%, 0.076%, 0.077%, 0.078%, 0.079%, 0.080%, 0.081%, 0
Hematopoietic stem cell frequency of .082%, 0.083%, 0.084%, 0.085%, or 0.086%. In some embodiments, the population of cells has a frequency of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells of about 0.066%.

In another aspect, the invention relates to a method of treating a stem cell disorder in a mammalian patient (e.g., a human patient), the method comprising treating a stem cell disorder in a mammalian donor (e.g., a human patient) according to any of the methods described above. , mobilizing a population of hematopoietic stem cells in a human donor) and infusing a therapeutically effective amount of hematopoietic stem cells or their progeny into the patient.

In a further aspect, the invention relates to a method of treating a stem cell disorder in a mammalian patient (e.g., a human patient), said method comprising a therapeutically effective stem cell disorder mobilized according to any of the methods described above. and injecting an amount of hematopoietic stem cells or their progeny into said patient.

In another aspect, the invention relates to a method of treating a stem cell disorder in a mammalian patient (e.g., a human patient), the method comprising administering any one or more of the pharmaceutical compositions described above to said patient. including administering to

In some embodiments of any of the three aspects, the stem cell disorder is a hemoglobinopathic disorder such as sickle cell anemia, thalassemia, Fanconi anemia, aplastic anemia, and Wiskott-Aldrich syndrome. be. In some embodiments, the stem cell disorder is a myelodysplastic disorder. The stem cell disorder may be an immunodeficiency disorder, such as a congenital immunodeficiency disorder or an acquired immunodeficiency disorder (eg, human immunodeficiency virus or acquired immunodeficiency syndrome). In some embodiments, the stem cell disorder is a metabolic disorder selected from, for example, glycogen storage disorders, mucopolysaccharidoses, Gaucher disease, Hurler disease, sphingolipidoses, and metachromatic leukodystrophy; It is.

In some embodiments, the stem cell disorder is cancer. The cancer may be, for example, leukemia, lymphoma, multiple myeloma, and neuroblastoma. In some embodiments, the cancer is a hematological cancer. In some embodiments, the cancer is acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, multiple myeloma,
Diffuse large B-cell lymphoma, or non-Hodgkin's lymphoma
ma).

In some embodiments, the stem cell disorder is adenosine deaminase deficiency and severe combined immunodeficiency, hyperimmune globulin M syndrome, Chediak-Higashi disease, hereditary lymphohistiocytosis, osteopetrosis, osteogenesis imperfecta. disease, storage disease, thalassemia major, systemic sclerosis, systemic lupus erythematosus, multiple sclerosis, and juvenile rheumatoid arthritis.

In some embodiments, the stem cell disorder is an autoimmune disorder, such as multiple sclerosis,
Treatment of human systemic lupus erythematosus, rheumatoid arthritis, inflammatory bowel disease, psoriasis, type 1 diabetes (1
type diabetes), acute disseminated encephalomyelitis, Addison's disease, generalized alopecia, ankylosing spondylitis, antiphospholipid antibody syndrome, aplastic anemia, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmunity Inner ear disease, autoimmune lymphoproliferative syndrome, autoimmune oophoritis, Barrow's disease, Behcet's disease,
Bullous pemphigoid, cardiomyopathy, Chagas disease, chronic fatigue immunodeficiency syndrome, chronic inflammatory demyelinating polyneuropathy, Crohn's disease, cicatricial pemphigoid, celiac sprue dermatitis herpetiformis
(coeliac sprue-dermatitis herpetiformis), cold agglutinin disease, CREST syndrome, Degos disease,
Discoid lupus erythematosus, autonomic neuropathy, endometriosis, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, Goodpasture syndrome, Graves' disease, Guillain-Barre syndrome (GBS), Hashimoto's thyroiditis, suppuration hidradenitis, idiopathic and/or acute thrombocytopenic purpura, idiopathic pulmonary fibrosis, IgA neuropathy, interstitial cystitis, juvenile arthritis, Kawasaki disease, lichen planus,
Lyme disease, Meniere's disease, mixed connective tissue disease, myasthenia gravis, neuromyotonia, opsoclonus-myoclonus syndrome, optic neuritis, ord's thyroiditis
), pemphigus vulgaris, pernicious anemia, polychondritis, polymyositis and dermatomyositis, primary biliary cirrhosis, polyarteritis nodosa, polyendocrine syndrome, polymyalgia rheumatica, primary agamma globulin primary agammaglobulinemia, Raynaud's phenomenon, Reiter's syndrome, rheumatic fever, sarcoidosis, scleroderma, Sjogren's syndrome, stiff person syndrome, Takayasu's arteritis, temporal arteritis, ulcerative colitis, uveitis, blood vessels vulvodynia, vitiligo, vulvodynia, and Wegener's granulomatosis.

In some embodiments, the hematopoietic stem cells are autologous to the patient. In some embodiments, the hematopoietic stem cells may be allogeneic to the patient, eg, HLA matched to the patient.

In some embodiments, the hematopoietic stem cells have been genetically modified to disrupt endogenous genes, such as genes encoding major histocompatibility complex proteins. The hematopoietic stem cells contain, for example, a CRISPR-related protein such as caspase 9, or a transcription activator-like effector nuclease.
, meganuclease, or another nuclease described herein, such as a zinc finger nuclease, may be genetically modified to destroy the endogenous gene.

In some embodiments, the hematopoietic stem cells or progeny thereof maintain their functional potential two or more days after infusion of the hematopoietic stem cells or progeny into the patient. In some embodiments, the hematopoietic stem cells or their progeny localize to hematopoietic tissue and/or reestablish hematopoiesis after injecting the hematopoietic stem cells or their progeny into the patient. In some embodiments, the hematopoietic stem cells or progeny thereof, when injected into the patient, are megakaryocytes, thrombocytes, platelets, red blood cells, mast cells, myoblasts, basophils, neutrophils. , eosinophils, microglia, granulocytes, monocytes, osteoclasts, and antigen presenting cells.

In another aspect, the invention relates to a method of mobilizing CD34 ^dim cells from the bone marrow of a human donor into peripheral blood, the method comprising: (i) a dose of about 50 μg/kg to about 1000 μg/kg of Gro -β, Gro-
a CXCR2 agonist selected from the group consisting of β T, and variants thereof; and (ii) C
XCR4 antagonist.

In another aspect, the invention relates to a method of performing allogeneic hematopoietic stem cell transplantation in a patient in need thereof, the method comprising infusing the patient with a therapeutically effective amount of allogeneic hematopoietic stem cells. wherein said hematopoietic stem cells are treated with (i) a CXCR2 agonist selected from the group consisting of Gro-β, Gro-β T, and variants thereof at a dose of about 50 μg/kg to about 1000 μg/kg; , and (ii) a CXCR4 antagonist, is mobilized from the human donor's bone marrow into the human donor's peripheral blood by a method comprising administering to said donor a CXCR4 antagonist.

In another aspect, the invention relates to a method of preventing, reducing the risk of developing, or reducing the severity of post-transplant infection in a patient in need thereof, the method providing therapeutically effective treatment to the patient in need thereof. injecting an amount of hematopoietic stem cells, wherein the hematopoietic stem cells are: (i) about 50 μl;
g/kg to about 1000 μg/kg of a CXCR2 agonist selected from the group consisting of Gro-β, Gro-β T, and variants thereof, and (ii) a CXCR4 antagonist to said donor. mobilization from the bone marrow of a human donor into the peripheral blood of a human donor.

In another aspect, the invention relates to a method of preventing, reducing the risk of developing, or reducing the severity of graft versus host disease (GVHD) in a patient in need thereof. , the method comprises injecting into the patient a therapeutically effective amount of hematopoietic stem cells, wherein the hematopoietic stem cells are treated with (i) Gro-β at a dose of about 50 μg/kg to about 1000 μg/kg; , Gro-β
a CXCR2 agonist selected from the group consisting of T, and variants thereof; and (ii) CXC
R4 antagonist is mobilized from the human donor's bone marrow into the human donor's peripheral blood by a method comprising administering to said donor an R4 antagonist.

In certain embodiments, the CD34 ^dim cells are present in greater numbers in peripheral blood than if the hematopoietic stem cells were mobilized using the CXCR4 antagonist alone. In certain embodiments, the CD34 ^dim cells are capable of inhibiting the proliferation of alloreactive T-lymphocytes when administered to a recipient.

In certain embodiments, the CXCR2 agonist is Gro-β T. In certain embodiments, the CXCR2 agonist is administered to the donor at a dose of about 100 μg/kg to about 250 μg/kg. In certain embodiments, the CXCR2 agonist is about 125 μg/kg to about 225 μg
/kg to said donor. In certain embodiments, the CXCR2 agonist is
A dose of approximately 150 μg/kg is administered to the donor. In certain embodiments, the CXCR2 agonist is administered intravenously to the donor.

In certain embodiments, said CXCR4 antagonist is administered subcutaneously to said donor.
In certain embodiments, the CXCR4 antagonist is plerixafor or a pharmaceutically acceptable salt thereof. In certain embodiments, said plerixafor, or a pharmaceutically acceptable salt thereof, is administered to said donor at a dose of about 50 μg/kg to about 500 μg/kg. In certain embodiments, the plerixafor or pharmaceutically acceptable salt thereof is present at a concentration of about 200 μg/kg to about
A dose of 300 μg/kg is administered to the donor. In certain embodiments, said plerixafor, or a pharmaceutically acceptable salt thereof, is administered to said donor at a dose of about 240 μg/kg.

In certain embodiments, the method further comprises testing a peripheral blood sample for the presence of CD34 ^dim cells and providing the sample for ex vivo expansion of the CD34 ^dim cells.

In certain embodiments, the invention relates to a population of CD34 ^dim cells derived from any of the above methods, or a composition comprising the same.

Figure 1A shows that when Gro-β T was administered to mice simultaneously with plerixafor (AMD3100), mobilization was synergistically increased and long-term hematopoietic stem cells (LT-HSC = Lin-c- This is a graph showing that the cells become enriched in kit ⁺ Sca-1 ⁺ CD150 ⁺ CD48 ⁺ ). Figure 1B shows that at 16 weeks, grafts containing cells recruited by Gro-β T and Plexaflor produced more competitive repopulating units ( FIG. 12 is a graph showing that the relative number of CRU) FIG. 2A is a graph showing the pharmacokinetic profile of Gro-β T at various doses when administered intravenously to rhesus monkeys. FIG. 2B is a graph showing the pharmacokinetic profile of various doses of Gro-β T when administered subcutaneously to rhesus monkeys. In both experiments, subjects received Gro-β T at the same time as plerixafor. FIG. 3A shows a series of graphs showing the recruitment response of white blood cells (white blood cells, "WBC") to various doses of Gro-β T when administered intravenously to rhesus monkeys. Leukocyte responses are shown in terms of both the amount of cells recruited (top) and the fold change in leukocyte density relative to pre-dose baseline leukocyte density (bottom). FIG. 3B shows a series of graphs showing the recruitment response of white blood cells (white blood cells, "WBC") to various doses of Gro-β T when administered subcutaneously to rhesus monkeys. Leukocyte responses are shown in terms of both the amount of cells recruited (top) and the fold change in leukocyte density relative to pre-dose baseline leukocyte density (bottom). In both experiments, subjects received Gro-β T at the same time as plerixafor. FIG. 4A shows a series of graphs showing the neutrophil recruitment response to various doses of Gro-β T when administered intravenously to rhesus monkeys. Neutrophil responses are shown in terms of both the amount of cells recruited (top) and the fold change in neutrophil density relative to pre-dose baseline neutrophil density (bottom). FIG. 4B shows a series of graphs showing the neutrophil recruitment response to various doses of Gro-β T when administered subcutaneously to rhesus monkeys. Neutrophil responses are shown in terms of both the amount of cells recruited (top) and the fold change in neutrophil density relative to pre-dose baseline neutrophil density (bottom). In both experiments, subjects received Gro-β T at the same time as plerixafor. FIG. 5A shows a series of graphs showing the lymphocyte recruitment response to various doses of Gro-β T when administered intravenously to rhesus monkeys. Lymphocyte responses are shown in terms of both the amount of cells recruited (top) and the fold change in lymphocyte density relative to baseline lymphocyte density before administration (bottom). FIG. 5B shows a series of graphs showing the lymphocyte recruitment response to various doses of Gro-β T when administered subcutaneously to rhesus monkeys. Lymphocyte responses are shown in terms of both the amount of cells recruited (top) and the fold change in lymphocyte density relative to baseline lymphocyte density before administration (bottom). In both experiments, subjects received Gro-β T at the same time as plerixafor. FIG. 6A shows a series of graphs showing the monocyte recruitment response to various doses of Gro-β T when administered intravenously to rhesus monkeys. Monocyte responses are shown in terms of both the amount of cells recruited (top) and the fold change in monocyte density relative to pre-dose baseline monocyte density (bottom). FIG. 6B shows a series of graphs showing monocyte recruitment responses to various doses of Gro-β T when administered subcutaneously to rhesus monkeys. Monocyte responses are shown in terms of both the amount of cells recruited (top) and the fold change in monocyte density relative to pre-dose baseline monocyte density (bottom). In both experiments, subjects received Gro-β T at the same time as plerixafor. FIG. 7A shows a series of graphs showing the recruitment response of CD34 ⁺ cells to various doses of Gro-β T when administered intravenously to rhesus monkeys. CD34 ⁺ cell responses are measured by the frequency of CD34 ⁺ cells in a sample obtained from the subject's peripheral blood (top) and the fold change in CD34 ⁺ cell frequency relative to the pre-dose baseline CD34 ⁺ cell frequency (bottom). Shown for both. FIG. 7B shows a series of graphs showing the recruitment response of CD34 ⁺ cells to various doses of Gro-β T when administered subcutaneously to rhesus macaques. CD34 ⁺ cell responses are measured by the frequency of CD34 ⁺ cells in a sample obtained from the subject's peripheral blood (top) and the fold change in CD34 ⁺ cell frequency relative to the pre-dose baseline CD34 ⁺ cell frequency (bottom). Shown for both. In both experiments, subjects received Gro-β T at the same time as plerixafor. FIG. 8A shows a series of graphs showing the recruitment response of CD34 ⁺ cells to various doses of Gro-β T when administered intravenously to rhesus monkeys. The CD34 ⁺ cell response is shown both in terms of the amount of cells recruited (top) and the fold change in CD34 ⁺ cell density relative to baseline CD34 ⁺ cell density before administration (bottom). Figure 8B shows a series of graphs showing the recruitment response of CD34 ⁺ cells to various doses of Gro-β T when administered subcutaneously to rhesus macaques. The CD34 ⁺ cell response is shown both in terms of the amount of cells recruited (top) and the fold change in CD34 ⁺ cell density relative to baseline CD34 ⁺ cell density before administration (bottom). In both experiments, subjects received Gro-β T at the same time as plerixafor. Figure 9A shows the effects of hematopoietic stem cells (CD34⁺CD90⁺ CD45RA^-Figure 2 shows a series of graphs illustrating the recruitment response of cells. CD34⁺ CD90⁺CD45RA^-Cellular responses to CD34 in samples obtained from the subject's peripheral blood⁺ CD90⁺CD45RA^-Cell frequency (top) and baseline/CD34 before administration⁺ CD90⁺CD45RA^-CD34 to cell frequency⁺ CD90⁺ CD45RA^-Shown both in terms of fold change in cell frequency (bottom). Figure 9B shows the response of various doses of Gro-β T to hematopoietic stem cells (CD34⁺CD90⁺ CD45RA^-Figure 2 shows a series of graphs illustrating the recruitment response of cells. CD34⁺ CD90⁺CD45RA^-Cellular responses to CD34 in samples obtained from the subject's peripheral blood⁺ CD90⁺CD45RA^-Cell frequency (top) and baseline/CD34 before administration⁺ CD90⁺CD45RA^- CD34 to cell frequency⁺ CD90⁺ CD45RA^-Shown both in terms of fold change in cell frequency (bottom). In both experiments, subjects received Gro-β T at the same time as plerixafor. FIG. 10A shows a series of graphs showing the recruitment response of hematopoietic stem cells (CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells) to various doses of Gro-β T when administered intravenously to rhesus monkeys. CD34 ⁺ CD90 ⁺ CD45RA ^- cell responses are measured by the amount of cells recruited (top) and the fold change in CD34 ⁺ CD90 ⁺ CD45RA ^- cell density relative to pre-dose baseline CD34 ⁺ CD90 ⁺ CD45RA ^- cell density (bottom). ). FIG. 10B shows a series of graphs showing the recruitment response of hematopoietic stem cells (CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells) to various doses of Gro-β T when administered subcutaneously to rhesus monkeys. CD34 ⁺ CD90 ⁺ CD45RA ^- cell responses are measured by the amount of cells recruited (top) and the fold change in CD34 ⁺ CD90 ⁺ CD45RA ^- cell density relative to pre-dose baseline CD34 ⁺ CD90 ⁺ CD45RA ^- cell density (bottom). ). In both experiments, subjects received Gro-β T at the same time as plerixafor. FIG. 11 shows a series of graphs demonstrating that intravenous administration of Gro-β T to rhesus monkeys at various doses increases the amount of colony forming units (CFU) of hematopoietic stem cells. CFR responses are shown both in terms of concentration of CFU (top) and fold change in CFU concentration relative to baseline CFU concentration before administration (bottom). In both experiments, subjects received Gro-β T at the same time as plerixafor. Figure 12A shows a series of graphs showing the response of plasma matrix metalloproteinase 9 (MMP9) to various doses of Gro-β T when administered intravenously to rhesus monkeys. Plasma MMP9 responses are shown both in terms of absolute concentration (top) and fold change in plasma MMP9 concentration relative to pre-dose baseline plasma MMP9 concentration (bottom). FIG. 12B shows a series of graphs showing plasma MMP9 responses to various doses of Gro-β T when administered subcutaneously to rhesus monkeys. Plasma MMP9 responses are shown both in terms of absolute concentration (top) and fold change in plasma MMP9 concentration relative to pre-dose baseline plasma MMP9 concentration (bottom). In both experiments, subjects received Gro-β T at the same time as plerixafor. FIG. 13A shows a series of graphs showing the response of tissue inhibitor of matrix metalloproteinases 1 (TIMP-1) to various doses of Gro-β T when administered intravenously to rhesus monkeys. Plasma TIMP-1 responses are shown both in terms of absolute concentration (top) and fold change in plasma TIMP-1 concentration relative to pre-dose baseline plasma TIMP-1 concentration (bottom). FIG. 13B shows a series of graphs showing the response of plasma TIMP-1 to various doses of Gro-β T when administered subcutaneously to rhesus monkeys. Plasma TIMP-1 responses are shown both in terms of absolute concentration (top) and fold change in plasma TIMP-1 concentration relative to pre-dose baseline plasma TIMP-1 concentration (bottom). In both experiments, subjects received Gro-β T at the same time as plerixafor. FIG. 14A shows a series of graphs showing the response of the molar ratio of plasma MMP9 to plasma TIMP-1 to various doses of Gro-β T when administered intravenously to rhesus monkeys. FIG. 14B shows a series of graphs showing the response of the molar ratio of plasma MMP9 to plasma TIMP-1 to various doses of Gro-β T when administered subcutaneously to rhesus monkeys. In both experiments, subjects received Gro-β T at the same time as plerixafor. Figure 15 is a typical flow plot of a blood sample taken from a rhesus macaque 4 hours after mobilization. Intravenous administration of 450 μg/kg Gro-β T and subcutaneous administration of 1 mg/kg plerixafor (AMD3100) mobilizes a population of CD34 ^dim cells. Figure 16 is a typical flow plot of a blood sample taken from a rhesus macaque at baseline versus a blood sample taken from a rhesus macaque 4 hours after mobilization. Mobilization was performed by (1) intravenous administration of 450 μg/kg Gro-β T and subcutaneous administration of 1 mg/kg plerixafor (AMD3100) or (2) subcutaneous administration of 1 mg/kg plerixafor (AMD3100). It was induced by Combining Gro-β T and plerixafor (compared to plerixafor alone) enhances the recruitment of CD34 ^dim cells. Figure 17 shows untreated rhesus monkeys (“non-mobilized”), rhesus monkeys treated with 450 μg/kg intravenous Gro-β T and 1 mg/kg plerixafor subcutaneously (“Gro-β T + plerixafor''), rhesus macaques treated with subcutaneous administration of 1 mg/kg plerixafor (``plerixafor''), and rhesus macaques treated with subcutaneous administration of 50 μg/kg (qd x 5) G-CSF (``G- 2 is a graph quantifying the concentration of CD34 ^dim cells in peripheral blood derived from ``CSF''. CD34 ^dim cells were present at significantly higher concentrations in blood mobilized using Gro-β T+plerixafor. FIG. 18 is a graph showing the composition of non-mobilized cells, G-CSF, Gro-β T and AMD3100, and grafts mobilized with AMD3100 alone. As shown, grafts mobilized using Gro-β T and AMD3100 compared to grafts mobilized using G-CSF. There is a 3-fold increase in CD34 ^dim cells and a 3-fold increase in T-cells. FIG. 19 shows that CD34 ^dim cells mobilized with Gro-β T and AMD3100 suppressed T-cell proliferation as measured by carboxyfluorescein succinimidyl ester (CFSE) staining after 4 days in culture. Here is the graph. "Beads" indicates that microbeads coated with anti-CD2/CD3/CD28 were used to stimulate T-cells. Figure 20 shows that by day 24, all mice (13/13) transplanted with unmobilized PBMC died of aGVHD, whereas 5/16 mice transplanted with peripheral blood mobilized with AMD3100, G - Survival curves showing that 3/16 mice transplanted with PBMCs mobilized with CSF died of aGVHD, and no mice transplanted with PBMCs mobilized with Gro-β T and AMD3100 died of aGVHD. It is. At day 60 post-transplant, 15/16 mice transplanted with Gro-β T and AMD3100 remained alive, while 10/16 mice transplanted with PBMCs mobilized with AMD3100 and PBMCs mobilized with G-CSF. Only 11/16 mice transplanted remained alive. p<0.0001 (comparing Gro-β T and AMD3100 with no recruitment), and p<0.05 (comparing Gro-β T and AMD3100 with AMD3100 alone). Figure 21A shows mice 14 days after transplantation with unmobilized PBMCs, PBMCs mobilized with Gro-β T and plerixafor (i.e., AMD3100), PBMCs mobilized with plerixafor alone, and PBMCs mobilized with G-CSF alone. The number of rhesus monkey CD45+CD3+ T-cells in Fig. 1 is shown. FIG. 21B shows 14 days after transplantation of unmobilized PBMCs, PBMCs mobilized with Gro-β T and plerixafor (i.e., AMD3100), and PBMCs mobilized with Gro-β T and plerixafor to remove CD34dim cells. The number of T-cells in mice is shown. FIG. 21C is a survival curve of mice transplanted with unmobilized PBMC, Gro-β T and AMD3100-mobilized PBMC, and Gro-β T and AMD3100-mobilized PBMC to remove CD34dim cells.

DETAILED DESCRIPTION The present invention provides compositions and methods for mobilizing hematopoietic stem and progenitor cells in a subject. For example, the subject may be a hematopoietic stem and progenitor cell donor (ie, a donor), such as a mammalian donor (eg, a human donor). The compositions and methods described herein may further be used to treat one or more stem cell disorders in a patient (eg, a human patient). Using the compositions and methods described herein, a CXC chemokine receptor type 2 (CXCR2) agonist (e.g., Gro-β or a variant thereof, e.g., a truncated form of Gro-β (e.g., Gro-β T )), optionally as described herein.
CXC chemokine receptor type 4 (CXCR4) antagonist (e.g. 1,1'-[1,4-phenylenebis(methylene)]-bis-1,4,8,11-tetra-azacyclotetradecane or variant thereof)
The donor may be administered with hematopoietic stem cells and an amount sufficient to mobilize the cells, as described herein. The compositions and methods described herein allow for the mobilization of hematopoietic stem and progenitor cells from the stem cell niche within the donor into circulating peripheral blood, while retaining leukocytes, neutrophils, lymphocytes, and monocytogenes. Recruitment of other cells of the hematopoietic lineage, such as spheres, can be reduced. Accordingly, the compositions and methods described herein allow selective mobilization of hematopoietic stem and progenitor cells in a donor, which can then be isolated from the donor for therapeutic use. Can be done.

The present invention relates, in part, to CXCR2 agonists, such as Gro-β, Gro-β T or variants thereof, and optionally to CXCR4 agonists, such as plerixafor or a pharmaceutically acceptable salt thereof.
Clinically important for mobilizing a population of cells enriched for hematopoietic stem cells compared to other cell types such as leukocytes, neutrophils, and monocytes when administered at specific doses in combination with antagonists It is based on the discovery that it can provide significant benefits. These other cell types are
may be undesirable for administration to human patients undergoing hematopoietic stem cell transplant therapy;
This effect is advantageous. Accordingly, populations of mobilized hematopoietic stem and progenitor cells produced using the compositions and methods described herein are particularly suitable for hematopoietic stem cell transplant therapy.

After mobilization, the hematopoietic stem or progenitor cells may be isolated for ex vivo expansion and/or for therapeutic use. In some embodiments, upon collection of mobilized hematopoietic stem and/or progenitor cells, the collected cells are transferred to a patient, e.g., a donor or another subject ( For example, the donor may be injected into a subject whose HLA is compatible with the donor. Additionally or alternatively, the mobilized cells are harvested and then contacted with an aryl hydrocarbon receptor antagonist to produce a population of hematopoietic stem cells having a sufficient amount of cells for transplantation. ex viv
May be grown in o.

As described herein, hematopoietic stem cells are capable of differentiating into multiple cell types of the hematopoietic lineage, thus allowing defective or missing cell types to be engrafted or re-engrafted in a patient. may be administered to the patient in order to Even if the patient is suffering from one or more blood diseases, such as, for example, an autoimmune disease, cancer, hemoglobinopathies, or other hematopoietic conditions, and is therefore in need of a hematopoietic stem cell transplant. good. Accordingly, the invention particularly relates to sickle cell anemia, thalassemia, Fanconi anemia, Wiskott-Aldrich syndrome, adenosine deaminase deficiency-severe combined immunodeficiency, metachromatic leukodystrophy, Diamond-Blackfan anemia, and Methods are provided for treating various hematopoietic conditions such as Schwachman-Diamond syndrome, human immunodeficiency virus infection and acquired immune deficiency syndrome, as well as cancer and autoimmune diseases.

The following section describes how to induce the recruitment of a population of hematopoietic stem or progenitor cells from the stem cell niche into the peripheral blood (subsequently isolate cells from this population, e.g.
CXCR4 may be administered to a donor to treat more than one type of stem cell disorder, such as cancer, autoimmune diseases, and injected into a patient to treat metabolic diseases described herein).
Antagonists and CXCR2 agonists are described. The following section further describes CXCR2
A method is described for determining whether a population of cells mobilized with an agonist and/or a CXCR4 antagonist is suitable for use in ex vivo expansion and/or for therapeutic applications.

DEFINITIONS As used herein, the term "about" refers to a value that is within 10% higher or lower than the stated value. For example, the term "about 5 nM" indicates a range of 4.5 nM to 5.5 nM.

As used herein, the terms "obtain" and "obtaining" mean acquiring possession of a physical entity or value, e.g. a numerical value, directly obtaining a physical entity or value. means to do or indirectly obtain. "Directly obtaining" means performing a process to obtain a physical entity or value (e.g., performing an assay or test on a sample, or analyzing a sample). do. "Obtaining indirectly" means receiving a physical entity or value from another party or source (e.g., a third party laboratory that directly obtained the physical entity or value). Directly obtaining a physical entity may include performing a process, e.g., collecting a sample (e.g., a single sample from a donor who has undergone or is undergoing a hematopoietic stem cell mobilization regimen as described herein). analyzing a sample of isolated hematopoietic cells). Obtaining values directly involves performing a process, e.g., an assay, on a sample or other object, e.g., the amount of hematopoietic stem cells in a sample, the amount of hematopoietic stem cells in a sample, the amount of hematopoietic stem cells on cells of another type of hematopoietic lineage, etc. Performing an analysis process that includes determining the proportion of stem cells or the frequency of hematopoietic stem cells in the total amount of cells in the sample.

As used herein, the term "affinity" refers to the affinity between two or more molecules, such as two or more proteins (e.g., a metalloproteinase and its endogenous inhibitor described herein). Refers to the strength of covalent interactions. The affinity, for example, as an equilibrium dissociation constant (K _d ),
Alternatively, when one of the binding partners is an enzyme, it can be expressed quantitatively as an inhibition constant (K _i ). Binding affinity may be measured using conventional techniques, such as enzyme-linked immunosorbent assays (ELISA), surface plasmon resonance assays, and isothermal titration calorimetry assays, among others.

As used herein, the term "antibody" refers to an immunoglobulin molecule that specifically binds to or is immunologically reactive with a particular antigen and may be polyclonal, monoclonal, including genetically engineered and other modified forms of antibodies, including, but not limited to, chimeric antibodies, humanized antibodies, heteroconjugate antibodies;
(e.g., bi-, tri-, and quad-specific antibodies, diabodies, triabodies, and tetrabodies), as well as antigen-binding fragments of antibodies (e.g., Fab', F(ab') ₂ , Fab
, Fv, rlgG, and scFv fragments). Unless otherwise specified, the term "monoclonal antibody" (mAb) refers to intact molecules and antibody fragments (including, for example, Fab and F(ab') ₂ fragments) that are capable of specifically binding to a target protein. This means that it includes both. As used herein, Fab and F(ab') ₂ fragments refer to antibody fragments that lack the Fc fragment of an intact antibody. Examples of these antibody fragments are described herein.

As used herein, the term "antigen-binding fragment" refers to one or more fragments of an antibody that retain the ability to specifically bind a target antigen. The antigen-binding function of antibodies can be performed by fragments of full-length antibodies. The antibody fragment may be, for example, a Fab, F(ab') ₂ , scFv, diabody, triabody, affibody, nanobody, aptamer, or domain antibody. Examples of binding fragments encompassed by the term "antigen-binding fragment" of an antibody include, but are not limited to: (i) Fab fragments, V _L , V _H , C _L and C _H Monovalent fragment consisting of one domain; (ii)F
(ab') ₂ fragments, a bivalent fragment comprising two Fab fragments, linked by a disulfide bridge in the hinge region; (iii) an Fd fragment consisting of the V _H and C _H 1 domains; (iv)
Fv fragments consisting of the V _L and V _H domains of a single arm of an antibody; (v) dAbs comprising V _H and V _L domains; (vi) dAb fragments consisting of V _H domains (e.g., Ward et al., Nature 341:544 -Five
46, (1989)); (vii) a dAb consisting of a V _H or V _L domain; (viii) an isolated complementarity determining region (CDR); and (ix) optionally linked by a synthetic linker. A combination of two or more (eg, two, three, four, five, or six) isolated CDRs that may be combined. Furthermore, although the two domains of the Fv fragment (V _L and V _H ) are encoded by separate genes, they can be joined by a linker using recombinant methods to form a single protein chain ( The V _L and V _H regions pair to form a monovalent molecule (known as a single chain Fv (scFv); see, for example, Bird et al., Science 242:423-426, ( (1988) and Huston et al., P
roc. Natl. Acad. Sci. USA 85:5879-5883, (1988)). These antibody fragments can be obtained using conventional techniques known to those skilled in the art, and the fragments can be screened for utility in the same manner as intact antibodies. Antigen-binding fragments can be produced by recombinant DNA techniques, enzymatic or chemical cleavage of intact immunoglobulins, or in some cases by chemical peptide synthesis procedures known in the art. .

As used herein, the term "bispecific antibody"
refers to, for example, a monoclonal, often human or humanized antibody capable of binding at least two different antigens.

As used herein, the term "CD34 ^dim cells" refers to a population of cells, at least a portion of which expresses the markers CD34, CD11b, and CD45, and which expresses the markers CD3, CD8, or CD20.
refers to a population of cells in which CD34 and CD45 are expressed at relatively low levels. This cell population exhibits monocyte properties, such as inhibiting the proliferation of alloreactive T-lymphocytes. (D'Aveni et al. (2015) Science Translational Medicine 7(281):1-12). In some embodiments, the population of CD34 ^dim cells may be CD14+.

Those skilled in the art will appreciate that CD34 ⁺ , the basal level of fluorescence (e.g., autofluorescence) in that cell population being tested and CD34 ⁺ bright cells (e.g., hematopoietic stem cells and/or CD34 ⁺ CD90 ⁺
Flow cytometry as a population of cells with a brightness between the brightness of the population
If a plot is visible, the CD34 ^dim cell population can be easily recognized. For example, in certain embodiments, the CD34 ^dim cell population is 5% of the CD34 ⁺ clear cell (e.g., hematopoietic stem cell) population.
~95% of the brightness, but brighter than the CD34 ⁻ cell population.

In certain embodiments, CD34 ^dim cells are 10 times brighter than CD34 ⁺ clear cells (e.g., hematopoietic stem cells).
% and 90%, 10% and 80%, 10% and 70%, 10% and 60%, 10% and 50%, 10% and 40%, 10% and 30%, 10% and 20%, 20
% and 90%, 20% and 80%, 20% and 70%, 20% and 60%, 20% and 50%, 20% and 40%, 20% and 30%, 30% and 90%, 30
% and 80%, 30% and 70%, 30% and 60%, 30% and 50%, 30% and 40%, 40% and 90%, 40% and 80%, 40% and 70%, 40
% and 60%, 40% and 50%, 50% and 90%, 50% and 80%, 50% and 70%, 50% and 60%, 60% and 90%, 60% and 80%, 60
% and 70%, 70% and 90%, 70% and 80%, or 80% and 90%, but brighter than CD34 ⁻ cells.
In certain embodiments, the CD34 ^dim cells are at least 5% brighter, at least 10% brighter, at least 20% brighter, or at least 30% brighter than CD34 ⁻ cells, but CD34 ⁺ bright cells (b
right cells) (e.g., hematopoietic stem cells).

In certain embodiments, CD34 ^dim cells can be grown using a flow system that uses magnetic beads instead of fluorescence.
Identification in cell samples by using cytometry. The magnetic beads selectively pull down CD34 ⁺ bright cells, leaving CD34 ^dim cells in the cell sample.

In certain embodiments, CD34 ^dim cells are identified by measuring the number of copies of CD34 that the cells express. For example, CD34 ^dim cells are CD34 ⁺ clear cells (e.g., hematopoietic stem cells)
may exhibit a CD34 copy number between 5% and 95% compared to CD34 ⁺ clear cells (e.g., hematopoietic stem cells), or 10% and 90%, 10% and 80% compared to CD34+ clear cells (e.g., hematopoietic stem cells) , 10% and 70%, 10% and 60%, 10% and 50%, 10% and
40%, 10% and 30%, 10% and 20%, 20% and 90%, 20% and 80%, 20% and 70%, 20% and 60%, 20% and 50%, 20% and
40%, 20% and 30%, 30% and 90%, 30% and 80%, 30% and 70%, 30% and 60%, 30% and 50%, 30% and 40%, 40% and
90%, 40% and 80%, 40% and 70%, 40% and 60%, 40% and 50%, 50% and 90%, 50% and 80%, 50% and 70%, 50% and
CD34 copy number between 60%, 60% and 90%, 60% and 80%, 60% and 70%, 70% and 90%, 70% and 80%, or 80% and 90% be.

As used herein, the term "complementarity determining region" (CDR) refers to hypervariable regions found in both the light and heavy chain variable domains of antibodies. The more highly conserved portions of variable domains are called framework regions (FR). The positions of the amino acids representing the hypervariable regions of antibodies are:
It may vary depending on the context and various definitions known in the art. Some positions within a variable domain may be considered hybrid hypervariable positions, meaning that these positions may be considered to be within a hypervariable region under one set of criteria, while another may be considered to be outside the hypervariable region under the set of criteria. One or more of these positions may also be found in extended hypervariable regions. The antibodies described herein are
These hybrid hypervariable positions may include modifications. Natural heavy and light chain variable domains each contain four backbone regions, primarily in a β-sheet structure, connected by three CDRs (CDRs connect the β-sheet structures and optionally forms part of it).
The CDRs in each chain are grouped closely together by the backbone region in the order FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, and together with CDRs from other antibody chains form the target binding site of the antibody. (Kabat et al., Sequences of Proteins of Immunological Interest, Nationa
l Institute of Health, Bethesda, MD, 1987). As used herein, unless otherwise specified, immunoglobulin amino acid residue numbering is performed according to the immunoglobulin amino acid residue numbering system of Kabat et al.

As used herein, in the context of administering one or more agents to a subject;
The term "end of administration" refers to the point at which one or more agents have been administered to a subject in their entirety. In some embodiments, the present invention includes a CXCR4 antagonist (e.g., plerixafor or a variant thereof) and/or a CXCR2 agonist (e.g., Gro-β or a variant thereof or a truncated form thereof, such as Gro-β T). The agents described in the book may be administered to a subject over a period of time, for example, by intravenous or subcutaneous injection. The drug is
"Administration has ended" when the prescribed dosage of said drug has been administered to the subject in its entirety.
considered to be. CXCR4 antagonists (e.g. plerixafor or variants thereof) and C
XCR2 agonists (e.g. Gro-β or variants thereof or truncated forms thereof, e.g.
ro-β T, etc.), the drugs are administered to the subject once the prescribed doses of all drugs of a particular regimen are administered to the subject in their entirety. Administration is considered complete.

As used herein, the term "conservative variation,""conservativesubstitution," or "conservative amino acid substitution" refers to similar physicochemical properties such as polarity, electrostatic charge, and steric volume. Refers to the substitution of one or more amino acids for one or more various amino acids. These properties are summarized for each of the 20 natural amino acids in Table 1 below.

⁺ Based on A3 volume: 50-100 small, 100-150 medium, 150-200 large, and >200 bulky.

From this table, conservative amino acid families include, for example, (i) G, A, V, L, P, and M, (
Includes ii) D and E, (iii) C, S and T, (iv) H, K and R, (v) N and Q, and (vi) F, Y and W. Thus, a conservative mutation or substitution is one that replaces an amino acid with a member of the same amino acid family (eg, replacing Ser with Thr or Lys with Arg).

As used herein, "CRU" (competitive repopulation unit)
g unit)) refers to a unit of measurement of long-term engrafting stem cells that can be detected after transplantation in vivo.

As used herein, the term "donor" refers to a subject, e.g., a mammalian subject (e.g., a human subject), from whom one or more cells have been isolated and whose cells, or progeny thereof, are (administered to recipient). The one or more cells may be, for example, a population of hematopoietic stem cells or progenitor cells.

As used herein, the term "diabody" refers to a bivalent antibody that contains two polypeptide chains, where each polypeptide chain has V _H and V _L on the same peptide chain. It comprises V _H and V _L domains connected by a linker that is so short that intramolecular association of the domains is not possible (eg, a linker consisting of 5 amino acids). With this configuration, each domain pairs with a complementary domain on another polypeptide chain to form a homodimeric structure. Thus, the term "triabody" refers to a trivalent antibody containing three peptide chains;
Each of them is linked by a linker (e.g., a linker consisting of 1-2 amino acids) that is too short to allow intramolecular association of the V _H and V _L domains within the same peptide chain.
Contains one V _H domain and one V _L domain. In order to fold into its native structure, peptides constructed in this way typically require the V _H domains and V _L domains of adjacent peptide chains.
trimerize so that the domains are spatially close to each other (e.g., Holliger et al., Proc. N
atl. Acad. Sci. USA 90:6444-48, 1993).

As used herein, the term "disrupt" with respect to a gene refers to preventing the formation of a functional gene product. A gene product is functional only if it performs its normal (wild type) function. Disruption of said gene prevents the expression of the functional element encoded by said gene and disrupts the expression of the functional element encoded by said gene and/or in the promoter and/or operator necessary for the expression of said gene in the animal. contains insertions, deletions, or substitutions of one or more bases. The gene to be disrupted may include, for example, removing at least a portion of the gene from the genome of said animal, modifying said gene to prevent expression of a functional factor encoded by said gene, interfering RNA, or foreign Dominant by gene
can be destroyed by the expression of negative factors. Materials and methods for genetically modifying hematopoietic stem/progenitor cells are detailed in US 8,518,701; US 2010/0251395; and US 2012/0222143, the disclosures of each of which are incorporated herein by reference in their entirety. (In case of conflict, the present specification will control).

Inactivating the gene to create a knockout animal and/or introducing the nucleic acid construct into the animal to create a founder animal and the knockout or nucleic acid construct using various techniques known in the art. Animal strains can be generated in which the construct is integrated into the genome. Such techniques include, but are not limited to, pronuclear microinjection (U.S. Pat. No. 4,873,191), retroviral-mediated gene transfer to the germline (Van der Putten et al. (1985) Proc. Natl. Acad. Sci. USA, 82:6148-6152), gene targeting to embryonic stem cells (Thompson et al. (1989) Cell, 56:313-321), embryo electroporation (Lo (1983) Mol. Cell Biol., 3:1803-1814), sperm-mediated gene transfer (Lavitrano et al. (2002) Proc. Natl. Acad. Sci. USA, 99:14230-14235; Lavitr
ano et al. (2006) Reprod. Fert. Develop., 18:19-23), somatic cells (e.g., cumulus cells or breast cells), or adult, fetal, or embryonic stem cells in vitro. Transformation followed by nuclear transfer (Wilmut et al. (1997) Nature, 385:810-813; and Wakayama et al. (1998)
Nature, 394:369-374). Pronuclear microinjection, sperm-mediated gene transfer, and somatic cell nuclear transfer are particularly useful techniques. A genome-modified animal is an animal whose genes have been modified in all of its cells, including its germline cells. When using methods to produce animals that are mosaic with respect to their genetic modification, the animals may be inbred and genomically modified progeny may be selected. For example, cloning may be used to create, for example, a mosaic animal (if the cell is modified at the blastocyst stage), or genomic modification may occur when a single cell is modified. Animals that have been modified not to sexually mature may be homozygous or heterozygous for that modification, depending on the specific approach used. If a specific gene is to be inactivated by knockout modification, homozygosity is usually required. Heterozygosity is often sufficient if a particular gene is to be inactivated by RNA interference or a dominant-negative strategy.

As used herein, "dual variable domain immunoglobulin"
"DVD-Ig" refers to an antibody that combines the target-binding variable domains of two monoclonal antibodies via a linker to create a tetravalent dual-targeting single agent (e.g., Gu et al. (2012) Meth. Enzymol., 502:25-41).

As used herein, the term "endogenous" refers to molecules, cells, tissues, or organs (e.g., hematopoietic stem cells, or megakaryocytes, platelets, etc.) naturally found in a particular organism, such as a human patient. (
thrombocytes), platelets, red blood cells, mast cells, myeloblasts, basophils, neutrophils,
eosinophils, microglial cells, granulocytes, monocytes, osteoclasts, antigen presenting cells, macrophages,
cells of the hematopoietic lineage such as dendritic cells, natural killer cells, T-lymphocytes, or B-lymphocytes).

As used herein, the term "engraftment potential" is used to refer to the ability of hematopoietic stem cells and hematopoietic progenitor cells to repopulate tissue, such that such cells naturally whether circulating or provided by transplantation. The term encompasses all events surrounding or leading to engraftment, such as tissue homing of cells and colonization of cells within the tissue of interest. The efficiency of engraftment or percentage of engraftment can be assessed or quantified using any clinically acceptable parameter known to those skilled in the art, for example, competitive repopulating unit (CRU). ); the uptake or expression of markers in tissues into which stem cells home, colonize, or engraft; or assess disease progression, hematopoietic stem cell and hematopoietic progenitor cell survival, or recipe This may include assessing the subject's progress depending on the patient's survival. Engraftment can also be determined by measuring the number of white blood cells in peripheral blood after transplantation. Engraftment can also be assessed by measuring the rate of recovery of bone marrow cells by donor cells in bone marrow aspirate samples.

As used herein, the term "exogenous" refers to molecules, cells, tissues, or organs (e.g., hematopoietic stem cells, or Megakaryocytes, thrombocytes, platelets, red blood cells, mast cells, myeloblasts, basophils, neutrophils, eosinophils, microglial cells, granulocytes, monocytes, osteoclasts, antigen-presenting cells ,
Cells of the hematopoietic lineage such as macrophages, dendritic cells, natural killer cells, T-lymphocytes, or B-lymphocytes) are described. Exogenous substances include those provided to an organism or to a culture extracted therefrom from an external source.

As used herein, the term "backbone region" or "FW region" includes amino acid residues proximal to the CDRs of an antibody, or antigen-binding fragment thereof. Residues of the FW region may be present, for example, in human antibodies, humanized antibodies, monoclonal antibodies, antibody fragments, Fab fragments, single chain antibody fragments, scFv fragments, antibody domains, and bispecific antibodies, among others. There are things to do.

As used herein, the term "hematopoietic progenitor cells" includes several cell types of the hematopoietic system, including granulocytes, monocytes, erythrocytes, megakaryocytes, B-cells and T-cells, among others. including, but not limited to, pluripotent cells capable of differentiating into different types of cells. Hematopoietic progenitor cells are committed to the hematopoietic cell lineage and generally do not self-renew. Hematopoietic progenitor cells can be identified, for example, by the expression pattern of cell surface antigens and include cells with the following immunophenotype: Lin ⁻ KLS ⁺ Flk2 ⁻ CD34 ⁺ . Hematopoietic progenitors include short-term hematopoietic stem cells, multipotent progenitors, common myeloid progenitors, granulocyte-monocytic progenitors, and megakaryocyte-erythroid progenitors.
The presence of hematopoietic progenitor cells can be determined functionally, e.g., colony forming unit cells, e.g. in a complete methylcellulose assay.
or phenotypically through the detection of cell surface markers using flow cytometry and cell sorting assays as described herein and known in the art. can.

As used herein, the term "hematopoietic stem cell"("HSC")
)) ability to self-renew and granulocytes (e.g. promyelocytes, neutrophils, eosinophils, basophils)
, red blood cells (e.g. reticulocytes, red blood cells), platelets (e.g. megakaryoblasts, platelets)
including producing megakaryocytes, platelets), monocytes (e.g., monocytes, macrophages), dendritic cells, microglia, osteoclasts, and lymphocytes (e.g., NK cells, B-cells, and T-cells). refers to immature blood cells that have the ability to differentiate into mature blood cells including, but not limited to, various lineages. Such cells may include CD34 ⁺ cells. CD34 ⁺ cells are immature cells that express the CD34 cell surface marker. In humans, CD34+ cells are thought to include a subpopulation of cells with stem cell properties defined above, whereas in mice, HSCs are CD34-
It is. Furthermore, HSC also refers to long-term repopulation HSC (LT-HSC) and short-term repopulation HSC (ST-HSC). L
T-HSC and ST-HSC are distinguished based on functional potential and expression of cell surface markers. For example, human HSCs contain CD34 ⁺ , CD38 ^- , CD45RA ^- , CD90 ⁺ , CD49F ⁺ , and lin ^- (CD2, CD3, CD
4, negative for mature lineage markers including CD7, CD8, CD10, CD11B, CD19, CD20, CD56, CD235A, etc.). In mice, bone marrow LT-HSCs are CD34-, SCA-1+, C-kit+, CD135-, Sl
amfl/CD150+, CD48-, lin- (negative for mature lineage markers including Ter119, CD11b, Gr1, CD3, CD4, CD8, B220, IL7ra, etc.), whereas ST-HSCs are CD34 ⁺ , SCA- 1 ⁺ , C-ki
t ⁺ , CD135 ^- , Slamfl/CD150 ⁺ , and lin ^- (Ter119, CD11b, Gr1, CD3, CD4, CD8, B220, IL7
Negative for mature lineage markers including ra, etc.). In addition, ST-HSCs are less quiescent and more proliferative than LT-HSCs under homeostatic conditions. However, LT-HSCs have a high self-renewal capacity (i.e., LT-HSCs can survive throughout adulthood and be serially transplanted through successive recipients), whereas ST-HSCs , have limited self-renewal ability (i.e., ST-H
SCs only survive for a limited period of time and cannot be serially transplanted). Any of these HSCs can be used in the methods described herein. ST-HSCs are particularly useful because they are highly proliferative and therefore can give rise to differentiated progeny more quickly.

As used herein, the term "functional potential of hematopoietic stem cells" refers to 1) pluripotency, which includes, but is not limited to, granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils), red blood cells (e.g. reticulocytes, red blood cells), platelets (e.g. megakaryoblasts, platelets)
t) producing megakaryocytes, platelets), monocytes (e.g. monocytes, macrophages), dendritic cells,
including microglia, osteoclasts, and lymphocytes (e.g. NK cells, B-cells and T-cells), etc.
2) self-renewal (this refers to the ability of a hematopoietic stem cell to give rise to daughter cells with a potential equal to that of the mother cell; furthermore, this ability becomes exhausted) 3) the ability of hematopoietic stem cells or their progeny to be reintroduced into the transplant recipient, home to the hematopoietic stem cell niche, and reestablish productive, sustained hematopoiesis. refers to the functional properties of hematopoietic stem cells, including

As used herein, the term "major histocompatibility complex antigen"("MHC", also referred to as "human leukocyte antigen" in humans) means a cell that has a unique antigenic identity. Refers to proteins expressed on the cell surface. MHC/HLA antigens can be expressed on T-cells, either as derived from the same hematopoietic stem cell source as the immune effector cells ("self"), or as derived from another source of hematopoietic reconstituted cells ("non-self"). and target molecules recognized by NK cells. Two main classes of HLA antigens are recognized: HLA class I and HLA class I
I. HLA class I antigens (A, B, and C in humans) enable each cell to be recognized as "self," whereas HLA class II antigens (DR, DP, and DQ in humans) are antigens that are associated with lymphocytes. Involved in the reaction between presenting cells. Both are believed to be involved in the rejection of transplanted organs. H.L.
An important aspect of the A gene system is its polymorphism. Each gene, MHC class I (A, B and C) and MHC
Class II (DP, DQ and DR) exists at different allelic loci. For example, two unrelated individuals may each carry the class I HLA-B genes B5 and Bw41. Allelic products differ by one or more amino acids in the alpha and/or beta domains. A large panel of specific antibodies or nucleic acid reagents is used to type individual HLA haplotypes using leukocytes expressing class I and class II molecules. Genes commonly used for HLA typing are the six MHC class I and class II proteins: HLA-A; HLA-B and HLA-A;
Two alleles for each of LA-DR. The HLA gene is located at chromosome position 6p21.
The six classically transplanted HLA genes, as well as the regulation of the immune system and several other fundamental molecular processes and cellular It encodes genes that encode at least 132 proteins that have important roles in regulating processes. The complete locus is approximately 3.6 Mb and contains at least 224
It has several genetic loci. One effect of this clustering is "haplotypes," ie, sets of alleles present on a single chromosome, which are inherited from one parent and tend to be inherited as a group. The set of alleles inherited from each parent is
They form haplotypes, in which several alleles tend to be associated together. Identifying a patient's haplotype can be useful in predicting the probability of finding a compatible donor and advancing search strategies. This is because some alleles and haplotypes are more common than others and are distributed at different frequencies in different racial and ethnic groups.

As used herein, the term "HLA matched" refers to any relationship between a donor and a recipient, such as a donor providing a hematopoietic stem cell graft to a recipient in need of hematopoietic stem cell transplantation therapy. HLA antigens also refer to donor-recipient pairs that are not incompatible. HLA compatible
Donor-recipient pairs (i.e., all six alleles matched) are less likely to have endogenous T-cells and NK cells recognizing the incoming graft as foreign, and thus have immunity to the graft. The risk of graft rejection is reduced because there is less chance of initiating a reaction.

As used herein, the term "HLA-mismatched" refers to providing a hematopoietic stem cell graft to a recipient in need of hematopoietic stem cell transplantation therapy, particularly with respect to HLA-A, HLA-B and HLA-DR. Refers to a donor-recipient pair, such as a donor, in which the donor and recipient are incompatible by at least one HLA antigen. In some embodiments, one haplotype is compatible and the other is incompatible. In HLA-mismatched donor-recipient pairs, endogenous T-cells and NK cells are more likely to recognize incoming grafts as foreign, and therefore
cells and NK cells are more likely to mount an immune response against the graft, potentially increasing the risk of graft rejection compared to an HLA-matched donor-recipient pair.

As used herein, the term "human antibody" refers to substantially all portions of a protein (e.g., all CDRs, backbone regions, C _L , C _H domains (e.g., C _H 1, C _H 2, C _H 3), hinge, and V _L and V _H domains) are substantially non-immunogenic in humans, with only minor sequence changes or mutations. Human antibodies can be prepared in human cells (e.g., by recombinant expression) or in functionally reconstituted human immunoglobulins (e.g., heavy and/or light chains).
) may be produced by non-human animals or prokaryotic or eukaryotic cells capable of expressing the gene. When a human antibody is a single chain antibody, it may contain a linker peptide not found in naturally occurring human antibodies. For example, an Fv may include a linker peptide, such as 2 to about 8 glycine or other amino acid residues, that connects the heavy chain variable region and the light chain variable region. Such linker peptides are considered to be of human origin. Human antibodies may be made by a variety of methods known in the art, including phage display methods using antibody libraries derived from human immunoglobulin sequences. Human antibodies are also unable to express functional endogenous immunoglobulins, but
They may also be produced using transgenic mice capable of expressing human immunoglobulin genes (e.g., PCT Publication No. WO 1998/24893; WO 1992/01047; WO 1996/34
096; WO 1996/33735; US Patent No. 5,413,923; No. 5,625,126; No. 5,633,425; No. 5,569,825
No. 5,661,016; No. 5,545,806; No. 5,814,318; No. 5,885,793; No. 5,916,771; and No. 5
, No. 939,598).

As used herein, the term "humanized" antibody refers to an antibody that contains minimal sequences derived from non-human immunoglobulins. Generally, a humanized antibody will contain substantially all of at least one, and typically two, variable domains in which all or substantially all of the CDR regions are those of a non-human immunoglobulin. handle. All or substantially all of the FW region may be that of a human immunoglobulin sequence. The humanized antibody may also include at least a portion of an immunoglobulin constant region (Fc), typically at least a portion of a human immunoglobulin consensus sequence. Methods for humanizing antibodies are known in the art and are described, for example, in Riechmann et al., Nature 332:323-7, 1988; US Pat. No. 5,530,101.
No. 5,585,089; No. 5,693,761; No. 5,693,762; and No. 6,180,370.

As used herein, a patient "in need of" a hematopoietic stem cell transplant includes a patient who exhibits a defect or deficiency in one or more blood cell types, as well as any stem cell disorder described herein. , including patients with autoimmune diseases, cancer, or other medical conditions. Hematopoietic stem cells are generally 1) multipotent and therefore include, but are not limited to, granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils), erythrocytes (e.g., reticulocytes); , red blood cells), platelets (e.g., megakaryoblasts, platelets (pla
megakaryocytes, platelets), monocytes (e.g., monocytes, macrophages), dendritic cells, microglia, osteoclasts, and lymphocytes (e.g., NK cells, B-cells, and T-cells). 2) capable of self-renewal, thus giving rise to daughter cells with a potential equal to that of the mother cell, and 3) capable of being repopulated into the transplant recipient. introduced and home to hematopoietic stem cell niches, demonstrating the ability to re-establish productive and sustained hematopoiesis. Accordingly, hematopoietic stem cells may be administered to a patient defective or deficient in one or more cell types of the hematopoietic lineage to reconstitute the defective or defective cell population in vivo. For example, the patient may be suffering from cancer and the defect may be caused by administering chemotherapeutic agents or other agents that selectively or non-specifically reduce cancerous cell populations. It may happen. Additionally or alternatively, the patient is suffering from a hemoglobinopathy (e.g., a non-malignant hemoglobinopathy) such as sickle cell anemia, thalassemia, Fanconi anemia, aplastic anemia, and Wiskott-Aldrich syndrome. There is. The subject may be a subject suffering from adenosine deaminase severe combined immunodeficiency disease (ADA SCID), HIV/AIDS, metachromatic leukodystrophy, Diamond-Blackfan anemia, and Shwachman-Diamond syndrome. . The subject has a genetic blood disorder (
may have or be affected by an autoimmune disease (for example, sickle cell anemia) or an autoimmune disease. Additionally or alternatively, the subject may have or be affected by a malignancy such as neuroblastoma or a hematological cancer. In some embodiments, the subject may have leukemia, lymphoma, or myeloma. In some embodiments, the subject has acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia,
Multiple myeloma, diffuse large B-cell lymphoma, or non-Hodgkin lymphoma
in's lymphoma). In some embodiments, the subject has myelodysplastic syndrome. In some embodiments, the subject has an autoimmune disease such as scleroderma, multiple sclerosis, ulcerative colitis, Crohn's disease, type I diabetes, or another autoimmune disease described herein. Have a pathological condition. In some embodiments, the subject is in need of chimeric antigen receptor T-cell (CART) therapy. In some embodiments, the subject has or is affected by a metabolic storage disorder. The subject has a metabolic disorder selected from glycogen storage disorders, mucopolysaccharidoses, Gaucher disease, Hurler disease, sphingolipidoses, metachromatic leukodystrophy, or would benefit from the treatments and therapies disclosed herein. Any other disease or disorder that may be present (including, but not limited to, severe combined immunodeficiency, Wiskott-Aldrich syndrome, hyperimmune globulin M (IgM) syndrome, Chediak-Higashi disease, hereditary lymphoma) Bone Marrow
"Transplantation for Non-Malignant disease", ASH Education Book, 1:319-338(2000)
(This disclosure relates to conditions that may be treated by applying hematopoietic stem cell transplantation therapy, and is incorporated herein by reference in its entirety). May be affected by or affected by the disease. Additionally or alternatively,
Patients ``in need'' of hematopoietic stem cell transplantation are those with or without one of the aforementioned conditions (nonetheless, those with megakaryocytes, thrombocytes, platelets)
et), red blood cells, mast cells, myeloblasts, basophils, neutrophils, eosinophils, microglia, granulocytes, monocytes, osteoclasts, antigen presenting cells, macrophages, dendritic cells, natural killer cells exhibiting decreased levels (e.g., compared to levels in other healthy subjects) of one or more endogenous cell types within the hematopoietic lineage, such as , T-lymphocytes, and B-lymphocytes. Sometimes. One of ordinary skill in the art will be able to determine the above cell types or other blood cell types by, for example, flow cytometry and fluorescence-activated cell sorting (FACS) methods, among other procedures known in the art. It can be easily determined whether the levels of one or more of these are reduced relative to otherwise healthy subjects.

As used herein, the term "leukocyte" refers to a heterogeneous group of nucleated blood cell types and excludes red blood cells and platelets. White blood cells are divided into two broad groups: polymorphonuclear cells, which include neutrophils, eosinophils, and basophils, and mononuclear cells, which include lymphocytes and monocytes. Polymorphonuclear cells contain many cytoplasmic granules and multilobed nuclei and include: neutrophils (generally amoeboid in shape, phagocytic, and stain with both basic and acidic dyes) )
, and eosinophils and basophils (containing cytoplasmic granules that stain with acidic and basic dyes, respectively).

As used herein, the term "lymphocyte" refers to a mononuclear leukocyte involved in initiating an immune response. Typically, lymphocytes include B-lymphocytes, T-lymphocytes, NK cells, and the like.

As used herein, the terms "mobilize" and "mobilization" refer to the process by which a population of hematopoietic stem or progenitor cells is released from a stem cell niche, such as the bone marrow of a subject, into circulation within the peripheral blood. means. Mobilization of hematopoietic stem and progenitor cells can be monitored, for example, by assessing the amount or concentration of hematopoietic stem or progenitor cells in a peripheral blood sample isolated from the subject. For example, after applying a hematopoietic stem or progenitor cell mobilization regimen to the subject, a peripheral blood sample is obtained from the subject and the amount or concentration of hematopoietic stem or progenitor cells in the peripheral blood sample is assessed. Sometimes. The mobilization regimen may include, for example, a CXCR4 antagonist as described herein (e.g., plerixafor or a variant thereof).
and the CXCR2 agonists described herein (e.g., Gro
-β or a variant thereof, eg, a truncated form of Gro-β, eg, Gro-β T). The amount or concentration of hematopoietic stem cells or progenitor cells in a peripheral blood sample isolated from said subject after applying a mobilization regimen; and the amount or concentration of hematopoietic stem cells in a peripheral blood sample isolated from said subject before applying a mobilization regimen. or the amount or concentration of progenitor cells. If an increase in the amount or concentration of hematopoietic stem cells or progenitor cells is observed in the peripheral blood of the subject after applying the mobilization regimen, then the subject is responsive to the mobilization regimen, and hematopoietic stem cells and that progenitor cells are being released into the peripheral blood circulation from one or more stem cell niches, such as the bone marrow. In some embodiments, after applying the mobilization regimen, the amount or concentration of hematopoietic stem or progenitor cells in the subject's peripheral blood is 1%, 100%, 1,000% or more (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%
, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%,
40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 200%, 300%, 40
0%, 500%, 600%, 700%, 800%, 900%, 1000% or more) indicates that the subject is responding to the mobilization regimen and that hematopoietic stem cells and This indicates that the progenitor cells were released into the peripheral blood circulation from one or more stem cell niches, such as the bone marrow. Methods for measuring the amount or concentration of hematopoietic stem or progenitor cells are described herein and known in the art, and include, for example, determining the amount or concentration of hematopoietic stem or progenitor cells according to the antigen expression profile of the cell. flow cytometry techniques (which are described herein). For example, human HSCs contain CD34 ⁺ , CD38 ⁻ , CD45RA ⁻ , CD90
⁺ , CD49F ⁺ , and lin-(CD2, CD3, CD4, CD7, CD8, CD10, CD11B, CD19, CD20, CD56, CD23
negative for mature lineage markers including 5A, etc.). Additional methods for determining the amount or concentration of hematopoietic stem or progenitor cells in a peripheral blood sample isolated from a subject include the number of colony forming units (CFU) in said sample, which is Included are assays that quantify the amount of viable hematopoietic stem or progenitor cells that, upon incubation in culture, give rise to individual populations of hematopoietic stem or progenitor cells.

As used herein, the term "mobilizing amount" refers to an amount that, when administered to a subject, e.g., a mammalian subject (e.g., a human subject), mobilizes a population of hematopoietic stem or progenitor cells.
an amount of one or more agents, e.g., a CXCR4 antagonist and/or CXCR as described herein;
2 agonist (in some embodiments, plerixafor or a variant thereof,
and/or Gro-β or a variant thereof, such as a truncated product of Gro-β, such as Gro-β T
amount). Exemplary mobilizing amounts of these agents include, for example, about 20 to about 40
CD34 ⁺ cells/μL peripheral blood, e.g., about 21 to about 39 CD34 ⁺ cells/μL peripheral blood, about 22 to about 38 CD34 ⁺
cells/μL peripheral blood, approximately 23 to approximately 37 CD34 ⁺ cells/μL peripheral blood, approximately 24 to approximately 36 CD34 ⁺ cells/μL peripheral blood
, approximately 25 to approximately 35 CD34 ⁺ cells/μL peripheral blood, approximately 26 to approximately 34 CD34 ⁺ cells/μL peripheral blood, approximately 27 to approximately 33 cells
CD34 ⁺ cells/μL peripheral blood, about 28 to about 32 CD34 ⁺ cells/μL peripheral blood, or about 29 to about 31 CD34 ⁺ cells
/μL peripheral blood (e.g. approximately 20 CD34 ⁺ cells/μL peripheral blood, 21 CD34 ⁺ cells/μL peripheral blood, 22 CD34 cells/μL peripheral blood
⁺ cells/μL peripheral blood, 23 CD34 ⁺ cells/μL peripheral blood, 24, CD34 ⁺ cells/μL peripheral blood, 25 CD34 ⁺ cells/μL peripheral blood, 26 CD34 ⁺ cells/μL peripheral blood, 27 CD34 ⁺ cells/μL peripheral blood, 28 CD34 ⁺ cells/μL
L peripheral blood, 29 CD34 ⁺ cells/μL peripheral blood, 30 CD34 ⁺ cells/μL peripheral blood, 31 CD34 ⁺ cells/μL peripheral blood, 32 CD34 ⁺ cells/μL peripheral blood 33 CD34 ⁺ cells/μL Peripheral blood, 34 CD34 ⁺ cells/μL peripheral blood, 35
CD34 ⁺ cells/μL peripheral blood, 36 CD34 ⁺ cells/μL peripheral blood, 37 CD34 ⁺ cells/μL peripheral blood, 38 CD3
⁴⁺ cells/μL peripheral blood, 39 CD34 ⁺ cells/μL peripheral blood, 40 CD34 ⁺ cells/μL peripheral blood, or more). For example, mobilizing amounts of CXCR2 agonists, such as Gro-β T, include from about 50 μg/kg to about 1 mg/kg, such as from about 50 μg/kg to about 300 μg/kg, from about 100 μg/kg to about 1 mg/kg, from about 50 μg/kg to about 300 μg/kg; μg/kg to about 250 μg/kg, or about 150 μg/kg. Mobilizing amounts of CXCR4 antagonists (e.g., plerixafor or a pharmaceutically acceptable salt thereof) may range from about 50 μg/kg to about 500 μg/kg to recipients (e.g., plerixafor or a pharmaceutically acceptable salt thereof).
from about 200 μg/kg to about 300 μg/kg, or about 240 μg/kg).

As used herein, the term "monoclonal antibody" refers to an antibody derived from a single clone, including any eukaryotic, prokaryotic, or phage clone, and the method by which it is produced. It does not originate from

As used herein, the term "monocyte" refers to CD14 ⁺ and CD34- peripheral blood mononuclear cells (per
ipheral blood mononuclear cell (PBMC), which generally refers to microbial products such as microbial products.
When activated by one or more foreign substances, they can differentiate into macrophages and/or dendritic cells. In particular, monocytes may have increased expression levels of the CD14 surface antigen marker, CD64,
CD93, CD180, CD328 (also known as sialic acid-binding Ig-like lectin 7 or Siglec7), and CD
329 (sialic acid-binding Ig-like lectin 9 or Siglec9), and peanut agglutinin protein (pean
ut agglutinin protein (PNA)).

As used herein, "peptide" refers to a single chain polyamide containing multiple amino acid residues, such as natural and/or non-natural amino acid residues, connected sequentially by amide bonds. Examples of peptides include shorter fragments of full-length proteins, such as full-length native proteins.

As used herein, the term "recipient" refers to a patient who receives a transplant, such as a transplant containing a population of hematopoietic stem cells. The transplanted cells administered to the recipient may be, for example, autologous,
It may be syngeneic or allogeneic.

As used herein, the term "sample" refers to a specimen collected from a subject (e.g., blood, blood components (e.g., serum or plasma), urine, saliva, amniotic fluid, cerebrospinal fluid, tissue (e.g., ,
placenta or dermis), pancreatic juice, chorionic villus samples, and cells). The sample can be, for example, peripheral blood collected from a donor who is undergoing or has undergone a hematopoietic stem or progenitor cell mobilization regimen as described herein.

As used herein, the term "scFv" refers to a single chain Fv antibody in which the variable domain of the heavy chain and the variable domain of the light chain from the antibody are joined to form a single chain. Point. The scFv fragment is
An antibody light chain variable region (V _L ) (e.g., CDR-L1, CDR-L2, and/or CDR-L3) and an antibody heavy chain variable region (V _H ) (e.g., CDR-L3) separated by a linker. H1, CDR-H2, and/or CDR-H
3) contains a single polypeptide chain. The linker connecting the V _L and V _H regions of the scFv fragment may be a peptide linker composed of protein-constituting amino acids. Alternative linkers may be used to increase the resistance of the scFv fragment to proteolytic degradation (e.g., linkers containing D-amino acids), to increase the solubility of the scFv fragment (e.g., polyethylene glycol-containing linkers or repeating glycine and hydrophilic linkers such as polypeptides containing serine residues) to improve the biophysical stability of said molecules (e.g. linkers containing cysteine residues that form intra- or intermolecular disulfide bonds). ), or to reduce the immunogenicity of the scFv fragment (eg, a linker containing a glycosylation site). Those skilled in the art will also appreciate that the variable regions of the scFv molecules described herein can be modified so as to vary in amino acid sequence from the antibody molecule from which they are derived. For example, nucleotide or amino acid substitutions that result in conservative substitutions or changes in amino acid residues (e.g., CDRs and / or in backbone residues).

As used herein, the phrase "stem cell disorder" refers to any disease, injury or Symptoms are broadly referred to as symptoms. For example, type I diabetes, among a variety of other disorders, has been shown to be cured by hematopoietic stem cell transplantation. Exemplary diseases that may be treated by infusing hematopoietic stem or progenitor cells into a patient include sickle cell anemia, thalassemia, Fanconi anemia, aplastic anemia, Wiskott-Aldrich syndrome, ADA.
These include SCID, HIV/AIDS, metachromatic leukodystrophy, Diamond-Blackfan anemia, and Shwachman-Diamond syndrome. Additional diseases that may be treated by transplanting hematopoietic stem or progenitor cells include blood disorders (e.g., sickle cell anemia) and scleroderma, multiple sclerosis, ulcerative colitis, and Crohn's disease. include autoimmune diseases.
Additional diseases that may be treated using hematopoietic stem and progenitor cell transplantation therapy include cancers, such as those described herein. Exemplary stem cell disorders include malignant tumors (
Examples include neuroblastoma, or blood cancers (eg, leukemia, lymphoma, and myeloma). In some embodiments, the cancer is acute myeloid leukemia, acute lymphocytic leukemia,
It may be chronic myeloid leukemia, chronic lymphocytic leukemia, multiple myeloma, diffuse large B-cell lymphoma, or non-Hodgkin's lymphoma. Additional diseases treatable using hematopoietic stem and progenitor cell transplantation therapy include myelodysplastic syndromes. In some embodiments, the patient has or is affected by a metabolic storage disorder. For example, the patient has a metabolic disorder selected from glycogen storage disorders, mucopolysaccharidoses, Gaucher disease, Hurler disease, sphingolipidoses, metachromatic leukodystrophy, or from the treatments and therapies disclosed herein. Any other disease or disorder that could benefit (including, but not limited to, Severe Combined Immunodeficiency, Wiskott-
Aldrich syndrome, hyperimmune globulin M (IgM) syndrome, Chediak-Higashi disease, hereditary lymphohistiocytosis, osteopetrosis, osteogenesis imperfecta, storage disease, thalassemia major, sickle cell disease, systemic sclerosis , systemic lupus erythematosus, multiple sclerosis, juvenile rheumatoid arthritis), and "Bone Marrow Transplantation for Non-Maligna"
nt disease", ASH Education Book, 1:319-338 (2000) (This disclosure relates to pathological conditions that may be treated by applying hematopoietic stem or progenitor cell transplantation therapy, and in its entirety. (herein incorporated by reference).

As used herein in the context of hematopoietic stem cell mobilization, the term "stem cell niche" refers to a mammalian donor (e.g., a human donor) in which endogenous hematopoietic stem or progenitor cells reside.
The microenvironment within the donor, such as An example of a stem cell niche is bone marrow tissue.

As used herein, the terms "subject" and "patient" refer to an organism, such as a human, being treated for a particular disease or condition as described herein. In some embodiments, a patient, such as a human patient in need of a hematopoietic stem cell transplant, is treated with a hematopoietic stem cell transplant to treat a stem cell disorder, e.g., a cancer, an autoimmune disease, or a metabolic disorder described herein. patients may receive treatments that include populations of hematopoietic stem cells. The hematopoietic stem cells transplanted into the patient may be a population of hematopoietic stem cells mobilized and harvested from a donor, eg, according to the compositions and methods described herein. In some embodiments, hematopoietic stem cells to be transplanted into the patient can be mobilized into the donor by administering a CXCR4 antagonist and/or CXCR2 agonist to the donor.

As used herein, the term "transfection" refers to methods of introducing foreign DNA into prokaryotic or eukaryotic host cells, such as electroporation, lipofection, calcium phosphate precipitation, DEAE-dextran transfection, etc. Any of a wide variety of commonly used techniques for deployment.

As used herein, the terms "treat" or "therapy" refer to therapeutic treatment, in which the purpose is to treat an undesirable physiological change or disorder in a patient undergoing treatment. or to promote beneficial phenotypes. The beneficial or desired clinical outcome is that following hematopoietic stem or progenitor cell transplantation therapy,
These include, but are not limited to, promoting the engraftment of foreign hematopoietic cells within a patient. In certain embodiments, the advantages include faster engraftment of transplanted cells (eg, neutrophils and platelets). For example, in certain embodiments, using the methods described herein, neutrophil recovery is achieved at about 5-20 days post-transplant, about 5-15 days post-transplant, about 5 days post-transplant. ~10 days, approximately 7 to 12 days after transplantation, approximately 8 to 12 days after transplantation, approximately 9 to 15 days after transplantation,
Occurs about 10 to 15 days after transplantation, or within about 10 days after transplantation. In certain embodiments, using the methods described herein, recovery of platelets occurs within about 10-20 days post-transplant, about 10-15 days post-transplant, about 15-15 days post-transplant. 20 days, about 12-18 days after transplant, about 12-17 days after transplant, about 13-18 days after transplant
Occurs within days, about 12 to 17 days after transplantation, or about 15 days after transplantation. A further beneficial result is that the cell number or relative concentration of hematopoietic stem cells in a patient in need of hematopoietic stem or progenitor cell transplantation is increased after administering the exogenous hematopoietic stem or progenitor cell graft to said patient. included. The beneficial results of the therapies described herein also include the use of one or more cells of the hematopoietic lineage (e.g., megakaryocytes, thrombocytes, platelets) after and subsequently to hematopoietic stem cell transplantation therapy.
telet), red blood cells, mast cells, myeloblasts, basophils, neutrophils, eosinophils, microglial cells, granulocytes, monocytes, osteoclasts, antigen-presenting cells, macrophages, dendritic cells, natural killers It may also include an increase in cell number or relative concentration of T-lymphocytes, T-lymphocytes, or B-lymphocytes. Additional beneficial results may include a reduction in the amount of disease-causing cell populations, such as cancer cells or autoimmune cell populations.

As used herein, the terms "variant" and "derivative" are used interchangeably and include naturally occurring compounds, peptides, proteins, or other substrates described herein. , synthetic, and semi-synthetic analogs. Variants or derivatives of compounds, peptides, proteins, or other substances described herein may retain or improve the biological activity of the original substance.

As used herein, the term "vector" includes nucleic acid vectors such as plasmids, DNA vectors, plasmids, RNA vectors, viruses, or other suitable replicons. The expression vectors described herein contain polynucleotide sequences as well as additional sequences used, for example, to express proteins and/or integrate these polynucleotide sequences into the genome of a mammalian cell. May contain elements. Particular vectors that can be used to express peptides and proteins, such as those described herein, include plasmids that contain regulatory sequences, such as promoter regions and enhancers that direct gene transcription. It will be done. Other useful vectors for expressing the peptides and proteins described herein enhance the translation rate of these genes or improve the stability or nuclear export of the mRNA resulting from gene transcription. containing a polynucleotide sequence. These sequence elements may include, for example, 5' and 3' untranslated regions and polyadenylation signal sites that ensure efficient transcription of the genes on the expression vector. Expression vectors described herein may also include polynucleotides encoding markers for selecting cells containing such vectors. Examples of suitable markers include genes encoding resistance to antibiotics such as ampicillin, chloramphenicol, kanamycin, and nourseothricin.

As used herein, the term "alkyl" refers to straight or branched chain alkyl groups having, for example, 1 to 20 carbon atoms in the chain. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, tert-pentyl, hexyl, isohexyl, and the like.

As used herein, the term "alkylene" refers to a straight or branched chain divalent alkyl group. The divalent positions may be on the same or different atoms within the alkyl chain. Examples of alkylene include methylene, ethylene, propylene, isopropylene, and the like.

As used herein, the term "heteroalkyl" includes, for example, 1 to 20 carbon atoms in the chain and one or more heteroatoms in the chain (e.g., oxygen, inter alia, A straight or branched alkyl group containing nitrogen or sulfur.

As used herein, the term "heteroalkylene" refers to a straight or branched chain divalent heteroalkyl group. The divalent positions may be on the same or different atoms within the heteroalkyl chain. A divalent position may be one or more heteroatoms.

As used herein, the term "alkenyl" refers to a straight or branched alkenyl group having, for example, 2 to 20 carbon atoms in the chain. Examples of alkenyl groups include vinyl, propenyl, isopropenyl, butenyl, tert-butylenyl, hexenyl, and the like.

As used herein, the term "alkenylene" refers to a straight or branched chain divalent alkenyl group. The divalent positions may be on the same or different atoms within the alkenyl chain. Examples of alkenylene include ethenylene, propenylene, isopropenylene, butenylene, and the like.

As used herein, the term "heteroalkenyl" has, for example, 2 to 20 carbon atoms in the chain and one or more heteroatoms in the chain, such as oxygen, inter alia, A straight or branched alkenyl group containing nitrogen or sulfur.

As used herein, the term "heteroalkenylene" refers to a straight or branched chain divalent heteroalkenyl group. The divalent positions may be on the same or different atoms within the heteroalkenyl chain. A divalent position may be one or more heteroatoms.

As used herein, the term "alkynyl" refers to a straight-chain or branched alkynyl group having, for example, 2 to 20 carbon atoms in the chain. Examples of alkynyl groups include propargyl, butynyl, pentynyl, hexynyl, and the like.

As used herein, the term "alkynylene" refers to a straight or branched chain divalent alkynyl group. The divalent positions may be on the same or different atoms within the alkynyl chain.

As used herein, the term "heteroalkynyl" has, for example, 2 to 20 carbon atoms in the chain and one or more heteroatoms in the chain (e.g., oxygen, inter alia, refers to a straight-chain or branched alkynyl group containing nitrogen or sulfur).

As used herein, the term "heteroalkynylene" refers to a straight or branched chain divalent heteroalkynyl group. The divalent positions may be on the same or different atoms within the heteroalkynyl chain. A divalent position may be one or more heteroatoms.

As used herein, the term "cycloalkyl" refers to monocyclic, or fused, bridged, or spiropolycyclic rings that are saturated and have, for example, 3 to 12 carbon ring atoms. Refers to a ring structure. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, bicyclo[3.1.0]hexane, and the like.

As used herein, the term "cycloalkylene" refers to a divalent cycloalkyl group. The divalent positions may be on the same or different atoms within the ring structure. Examples of cycloalkylene include cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, and the like.

As used herein, the term "heterocycloalkyl" is saturated and selected from heteroatoms, e.g., made of carbon atoms and selected from, e.g., nitrogen, oxygen, and sulfur, among others. monocyclic, or fused, bridged, or spiropolycyclic ring structures having from 3 to 12 ring atoms per ring structure. The ring structure may include one or more oxo groups, for example on carbon, nitrogen, or sulfur ring members.

As used herein, the term "heterocycloalkylene" refers to a divalent heterocycloalkyl group. The divalent positions may be on the same or different atoms within the ring structure.

As used herein, the term "aryl" refers to a monocyclic or polycyclic aromatic ring system containing, for example, 6 to 19 carbon atoms. Aryl groups include phenyl, fluorenyl,
These include, but are not limited to, naphthyl and the like. A divalent position may be one or more heteroatoms.

As used herein, the term "arylene" refers to a divalent aryl group. The divalent positions may be on the same atom or on different atoms.

As used herein, the term "heteroaryl" refers to a monocyclic heteroaromatic, or a bicyclic or tricyclic fused ring heteroaromatic group. Heteroaryl groups include pyridyl, pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,3,4-triazinyl, 1,2,3-triazinyl, benzofuryl, [2,3-dihydro]benzofuryl , isobenzofuryl, benzothienyl, benzotriazolyl, isobenzothienyl, indolyl, isoindolyl, 3H-indolyl, benzimidazolyl, imidazo[1,2-
a]Pyridyl, benzothiazolyl, benzoxazolyl, quinolidinyl, quinazolinyl, phthalazinyl, quinoxalinyl, cinnolinyl, naptyridinyl, pyrido[3,4-b]pyridyl, pyrido[3,2-b]pyridyl, pyrido[4,3-b ]Pyridyl, quinolyl, isoquinolyl, tetrazolyl, 5,6,7,8-tetrahydroquinolyl, 5,6,7,8-tetrahydroisoquinolyl, purinyl,
Examples include pteridinyl, carbazolyl, xanthenyl, benzoquinolyl and the like.

As used herein, the term "heteroarylene" refers to a divalent heteroaryl group. The divalent positions may be on the same atom or on different atoms. A divalent position may be one or more heteroatoms.

Unless otherwise constrained by the definition of the individual substituents, the chemical moieties described above, e.g., "alkyl", "alkylene", "heteroalkyl", "heteroalkylene", "
"alkenyl", "alkenylene", "heteroalkenyl", "heteroalkenylene", "
"alkynyl", "alkynylene", "heteroalkynyl", "heteroalkynylene",
"Cycloalkyl", "cycloalkylene", "heterocycloalkyl", "heterocycloalkylene", "aryl", "arylene", "heteroaryl", "heteroarylene" groups, etc., optionally include May be replaced. As used herein, the term "optionally substituted" refers to one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9
, 10, or more) substituents, as permitted by the valency of said compound or moiety or its moieties, such as alkyl, alkenyl, alkynyl, cycloalkyl, hetero Cycloalkyl, alkylaryl, alkylheteroaryl, alkylcycloalkyl, alkylheterocycloalkyl, amino, ammonium, acyl, acyloxy, acylamino, aminocarbonyl, alkoxycarbonyl, ureido, carbamate, aryl, heteroaryl, sulfinyl, sulfonyl, alkoxy, Refers to a compound or moiety containing a substituent selected from the group consisting of sulfanyl, halogen, carboxy, trihalomethyl, cyano, hydroxy, mercapto, nitro, and the like. The substitution is such that adjacent substituents undergo ring closure, e.g., ring closure of adjacent functional substituents, e.g., lactams, lactones, cyclic anhydrides, acetals, hemiacetals, thioacetals, aminals, and hemiaminals. for example, to provide a protecting group.

Methods for mobilizing hematopoietic stem and progenitor cells and providing cells for proliferation and therapeutic use
The present invention provides a method of administering to a mammalian donor (e.g., a human donor) a specific dose of Gro-β, Gro-β T, or a variant thereof, optionally in combination with a CXCR4 antagonist. CX
Administration of CR2 agonists can mobilize hematopoietic stem and progenitor cells, while reducing the recruitment of other cell types such as leukocytes, neutrophils, lymphocytes and monocytes. It is based, in part, on the discovery that it is possible. This property is particularly advantageous in the context of hematopoietic stem cell transplant therapy. Because hematopoietic stem cells mobilized and isolated from donors using the compositions and methods described herein reduce the amount of cell types that are undesirable to administer to human patients suffering from stem cell disorders. It is.

In particular, a CXCR2 agonist such as Gro-β, Gro-β T, or a variant thereof is approximately 50 μg/
kg to about 1 mg/kg, preferably about 100 μg/kg to about 250 μg/kg, even more preferably about 150 μg
donor (e.g., a mammalian donor, such as a human donor) when administered intravenously at a dose of /kg.
) to rapidly mobilize hematopoietic stem and progenitor cells, while it may be undesirable to infuse them into patients undergoing hematopoietic stem cell transplantation therapy (e.g., mammalian patients, such as human patients). It has been discovered that this reduces the recruitment of other cells in the lineage. A CXCR2 agonist, e.g., Gro-β, Gro-β T, or a variant thereof, when administered to a donor at the above doses, has the ability to selectively mobilize hematopoietic stem cells, as detailed in Example 1 below. shows.

mobilized in the donor by administering a CXCR2 agonist such as Gro-β, Gro-β T, or a variant thereof, and optionally a CXCR4 antagonist such as plerixafor or a pharmaceutically acceptable salt thereof. one or more of the ones listed in Table 2, characterizing a sample of said donor's peripheral blood in determining whether the hematopoietic stem cells are suitable for ex vivo expansion and/or for serving for therapeutic use. may obtain input values for each of the parameters. One or more parameters may be compared to a corresponding reference standard for each parameter, and if said reference standard is met by the sample of hematopoietic stem cells, the cells isolated from said donor are It may be provided for propagation and/or for infusion into a patient for therapeutic use (eg, to treat one or more stem cell disorders described herein).

Exemplary hematopoietic stem cell parameters and corresponding reference standards useful in conjunction with the compositions and methods described herein are shown in Table 2 below.

To determine whether a population of hematopoietic stem cells obtained from a donor (e.g., a mammalian donor, such as a human donor) is suitable for expansion ex vivo or for serving for therapeutic use. In selecting parameters, one or more of the input parameters listed in Table 2 may be selected. In some embodiments, individual parameters may be selected from parameter numbers 1-21. Alternatively, a proportion parameter (e.g., one or more of parameter numbers 1-17 in Table 2) and a frequency parameter (e.g., one or more of parameter numbers 18-21 listed in Table 2) of CD34 ⁺ cells. You may also select a combination of parameters such as: In some embodiments, the population of hematopoietic stem cells obtained from a donor (e.g., a mammalian donor, such as a human donor) is suitable for expansion ex vivo or for use in therapeutic applications. The parameters used to determine whether there are

CXCR2 agonist
Gro-β, Gro-β T and variants thereof Exemplary CXCR2 agonists that can be used with the compositions and methods described herein are Gro-β and variants thereof. Gro-β (growth regulatory protein β, chemokine (C-
XC motif) ligand 2 (CXCL2), and macrophage inflammatory protein 2-α (MIP2-α)), for example, by stimulating the release of proteases, particularly MMP9, from peripheral neutrophils. It is a cytokine that can recruit hematopoietic stem cells and progenitor cells. Although not limited by mechanism, MMP9 is associated with proteins such as stem cell factor, its corresponding receptor, CD117, and CXCL12, all of which commonly immobilize hematopoietic stem and progenitor cells in the bone marrow. Stimulating the degradation of hematopoietic stem and progenitor cells from cellular niches such as bone marrow into circulating peripheral blood may induce hematopoietic stem and progenitor cells to be mobilized from cellular niches such as bone marrow into circulating peripheral blood.

In addition to Gro-β, exemplary CXCR2 agonists that can be used with the compositions and methods described herein have 1 to 8 amino acids deleted at the N-terminus of Gro-β. (e.g., a peptide characterized by an N-terminal deletion of 1 amino acid, 2 amino acids, 3 amino acids, 4 amino acids, 5 amino acids, 6 amino acids, 7 amino acids, or 8 amino acids) ) is a truncated form of Gro-β. In some embodiments, a CXCR2 agonist that can be used with the compositions and methods described herein is characterized by having the first four amino acids deleted from the N-terminus of Gro-β. Contains Gro-β T. Gro-β T exhibits particularly advantageous biological properties, such as its ability to induce recruitment of hematopoietic stem and progenitor cells with several orders of magnitude greater potency than Gro-β. Gro-β and Gro-β T are, for example, US Patent No. 6,08
No. 0,398, the disclosure of which is incorporated herein by reference in its entirety.

Additionally, exemplary CXCR2 agonists that can be used with the compositions and methods described herein include aspartic acid in place of the asparagine residue at position 69 of SEQ ID NO: 1. It is a variant of Gro-β containing residues. This peptide, referred to herein as Gro-β N69D, retains the ability of Gro-β to recruit hematopoietic stem and progenitor cells, but induces this effect with greater potency. Similarly, CXCR2 agonists that can be used with the compositions and methods described herein include an aspartic acid residue in place of the asparagine residue at position 65 of SEQ ID NO: 2. Variants of Gro-β T containing groups are included. This peptide, referred to herein as Gro-β T N65D, not only retains the ability to recruit hematopoietic stem and progenitor cells, but also exerts significantly greater potency than that of Gro-β T. For example, Gro-β N69D and Gro-β T N65D are described in US Pat. No. 6,447,766, the disclosure of which is incorporated herein by reference in its entirety.

The amino acid sequences of Gro-β, Gro-β T, Gro-β N69D and Gro-β T N65D are listed in Table 7 below.

Additional CXCR2 agonists that can be used with the compositions and methods described herein include other variants of Gro-β, such as one or more amino acid substitutions compared to Gro-β;
Includes peptides with insertions and/or deletions. In some embodiments, a CXCR2 agonist that can be used with the compositions and methods described herein has SEQ ID NO.
ID NO: 1 (e.g., at least 85%, 90%, 95%, 96 %、97%
, 98%, 99%, 99.5%, or 100% sequence identity). In some embodiments, the amino acid sequence of the CXCR2 agonist differs from the amino acid sequence of SEQ ID NO: 1 only by one or more conservative amino acid substitutions. In some embodiments, CXCR
The amino acid sequence of the 2 agonist differs from SEQ ID NO. (S
EQ ID NO): Different from the amino acid sequence of 1. In some embodiments, the amino acid sequence of the CXCR2 agonist has no more than 20, no more than 15, no more than 10, no more than 5, or no more than 1 non-conservative amino acid substitution, such as SEQ ID NO: The amino acid sequence differs from that of 1.

Additional examples of CXCR2 agonists useful in conjunction with the compositions and methods described herein include G.
Gr such as peptides having one or more amino acid substitutions, insertions, and/or deletions relative to ro-β T
It is a variant of o-β T. In some embodiments, the CXCR2 agonist is a peptide (
For example, at least 85%, 90%, 95%, 96% for the amino acid sequence of SEQ ID NO: 2
, 97%, 98%, 99%, 99.5%, or 100% sequence identity).
In some embodiments, the amino acid sequence of the CXCR2 agonist differs from the amino acid sequence of SEQ ID NO: 2 only by one or more conservative amino acid substitutions. In some embodiments, the CXCR2 agonist has an amino acid sequence of 20 or less, 15 or less, 10 or less, 5
differs from the amino acid sequence of SEQ ID NO: 2 by less than or equal to 1 non-conservative amino acid substitution.

Additional examples of CXCR2 agonists useful in conjunction with the compositions and methods described herein include G.
A variant of ro-β N69D, such as a peptide having one or more amino acid substitutions, insertions, and/or deletions relative to Gro-β N69D. In some embodiments, the CXCR2 agonist is a peptide having at least 85% sequence identity to the amino acid sequence of SEQ ID NO: 3 (e.g., the amino acid sequence of SEQ ID NO: 3). At least 8 for the amino acid sequence
5%, 90%, 95%, 96%, 97%, 98%, 99%, 99.%, or 100% sequence identity). In some embodiments, the amino acid sequence of the CXCR2 agonist differs from the amino acid sequence of SEQ ID NO: 3 only by one or more conservative amino acid substitutions. In some embodiments, the amino acid sequence of the CXCR2 agonist has no more than 20, no more than 15, no more than 10, no more than 5, or no more than 1 non-conservative amino acid substitution, such as SEQ ID NO.
ID NO): Different from the amino acid sequence of 3.

Additional examples of CXCR2 agonists useful in conjunction with the compositions and methods described herein include G.
Variants of ro-β T N65D, e.g., one or more amino acid substitutions for Gro-β T N65D;
A peptide with insertions and/or deletions. In some embodiments, said CXCR2
The agonist is a peptide having at least 85% sequence identity to the amino acid sequence of SEQ ID NO: 4 (e.g., at least 85% to the amino acid sequence of SEQ ID NO: 4; 90%, 95%, 96%, 97%, 98%, 99%, 99.%, or 100% sequence identity). In some embodiments, the amino acid sequence of the CXCR2 agonist differs from the amino acid substitutions of SEQ ID NO: 4 only by one or more conservative amino acid substitutions. In some embodiments, the amino acid sequence of the CXCR2 agonist has no more than 20, no more than 15, no more than 10, no more than 5, or no more than 1 non-conservative amino acid substitution, such as SEQ ID NO. : Different from the amino acid sequence of 4.

Agonistic Anti-CXCR2 Antibodies and Antigen-Binding Fragments Thereof In some embodiments, the CXCR2 agonist is an antibody or antigen-binding fragment thereof that binds CXCR2 and activates CXCR2 signaling. In some embodiments,
The CXCR2 agonist, for example, when evaluated by a competitive CXCR2 binding assay,
It may be an antibody or an antigen-binding fragment thereof that binds to approximately the same epitope on CXCR2 that Gro-β or a variant or truncated product thereof (eg, Gro-β T, etc.) binds. In some embodiments, the CXCR2 agonist is an antibody or antigen-binding fragment thereof that competes with Gro-β or a variant or truncated form thereof (eg, Gro-β T, etc.) for binding to CXCR2.

In some embodiments of any of the above aspects, the antibody or antigen-binding fragment thereof is a monoclonal antibody or antigen-binding fragment thereof, a polyclonal antibody or antigen-binding fragment thereof, a humanized antibody or antigen-binding fragment thereof, bispecific antibodies or antigen-binding fragments thereof, dual-variable immunoglobulin domains, single-chain Fv molecules (scFv), diabodies, triabodies, nanobodies, antibody-like protein scaffolds; Fv fragment, Fab fragment, F(ab') ₂ molecule, and tandem di-scFv.
In some embodiments, the antibody has an isotype selected from the group consisting of IgG, IgA, IgM, IgD, and IgE.

Synthetic CXCR2 agonist
Peptide CXCR2 agonists described herein, such as Gro-β, Gro-β T, and variants thereof, can be synthesized using, for example, solid phase peptide synthesis techniques. Systems and processes for performing solid phase peptide synthesis are known in the art, and include, for example, U.S. Pat. (herein incorporated by reference) as it relates to protocols and techniques for the synthesis of peptides on-body. Solid-phase peptide synthesis involves adding amino acid residues to a peptide immobilized on a solid support, such as a polymeric resin (e.g., a hydrophilic resin such as a polyethylene-glycol-containing resin, or a hydrophobic resin such as a polystyrene-based resin). It is a process of adding groups.

Peptides, especially those containing protecting groups on amino, hydroxy, thiol, and carboxy substituents, may be attached to a solid support such that the peptide is effectively immobilized on the solid support. be converted into For example, the peptides may be attached to the support via their C-terminus, thereby immobilizing the peptides for subsequent reaction at the resin-liquid interface.

The process of adding amino acid residues to an immobilized peptide includes exposing at least some protecting groups to the immobilized peptide to remove at least some of the protecting groups from at least some of the immobilized peptide. Sometimes. The step of exposing to a deprotecting reagent may be configured such that, for example, side chain protecting groups are preserved while N-terminal protecting groups are removed. For example, exemplary amino protection includes a fluorenylmethyloxycarbonyl (Fmoc) substituent. A deprotection reagent containing a strongly basic substance such as piperidine (e.g., a solution of piperidine in a suitable organic solvent such as dimethylformamide (DMF)) is exposed to the immobilized peptide to
The moc protecting group may be removed from at least a portion of the immobilized peptide. Other protecting groups suitable for protecting amino substituents include, for example, tert-butyloxycarbonyl (Boc) moieties. A deprotection reagent containing a strong acid, such as trifluoroacetic acid (TFA), is exposed to an immobilized peptide containing a Boc-protected amino substituent to remove the Boc by an ionization process.
Protecting groups may be removed. In this way, the peptide can be protected and deprotected at specific positions (e.g., at one or more side chains or at the N-terminus or C-terminus of the immobilized peptide),
Chemical functional groups may be added regioselectively to one or more of these positions. This may be used, for example, to derivatize the side chains of immobilized peptides or to synthesize peptides, for example from the C-terminus to the N-terminus.

The process of adding amino acid residues to an immobilized peptide may include, for example, exposing a protected activated amino acid to an immobilized peptide such that at least a portion of said activated amino acid binds to said immobilized peptide and generates a new may include forming an amino acid residue attached to. For example, the peptide may be exposed to an activated amino acid that reacts with the deprotected N-terminus of the peptide, extending the peptide chain by one amino acid. An amino acid can be activated to react with a deprotected peptide by reacting the amino acid with an agent that enhances the electrophilicity of the backbone carbonyl carbon of the amino acid. For example, phosphonium and uronium salts can convert protected amino acids into activated species (e.g. , BOP, PyBOP, HBTU, and TBTU all produce HOBt esters). Other reagents can be used to help prevent racemization that may be induced in the presence of base. These reagents include auxiliary nucleophiles such as 1-hydroxy-benzotriazole (HOBt), 1-hydroxy-azabenzotriazole (HOAt), or
HOSu) or derivatives thereof (eg, DCC or WSCDI). Another reagent that can be utilized to prevent racemization is TBTU. Isobutyl
Mixed anhydride methods using chloroformates, with or without added auxiliary nucleophiles, can also be used similarly to azide methods due to the low racemization associated with this reagent. be. These types of compounds may also increase the rate of carbodiimide-mediated coupling, as well as prevent dehydration of Asn and Gln residues. Typical additional reagents also include bases such as N, N-diisopropylethylamine (DIPEA), triethylamine (TEA) or N-methylmorpholine (NMM). These reagents are described, for example, in U.S. Pat.
, 546,350, the disclosure of which is incorporated herein in its entirety.

During recombinant expression and folding of Gro-β and Gro-β T in aqueous solution, the C-terminal asparagine residues (Asn69 in Gro-β and Asn65 in Gro-β T) are specifically deamidated. easy to receive. This process results in the conversion of asparagine residues to aspartic acid. Without wishing to be bound by any theory, the chemical synthesis of Gro-β and Gro-β T is possible by providing conditions that reduce the exposure of this asparagine residue to nucleophilic solvents, e.g. , this problem can be overcome. For example, when prepared synthetically (e.g., chemically) using the solid-phase peptide synthesis techniques described above, synthetic Gro-β can be used in combination with the compositions and methods described herein. , Gro-β T, and their variants may, for example, have a purity of at least about 95% relative to deamidated versions of these peptides (i.e., less than 5% of the corresponding deamidated peptide ). for example,
Synthetic Gro-β, Gro-β, which may be used in conjunction with the compositions and methods described herein
β T, and their variants are deamidated versions of these peptides (e.g.
Asn69 deamidated version of SEQ ID NO: 1 or Asn of SEQ ID NO: 2
65 deamidated version), approximately 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5
%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99% or higher. For example, synthetic Gro-β, Gro-β T, and their variants can be synthesized by, for example, deamidated versions of these peptides (e.g., Asn69 deamidated versions of SEQ ID NO: 1 or SEQ ID NO: SEQ ID NO): For the Asn65 deamidated version of 2), approx.
95% to about 99.99% purity, such as about 95% to about 99.99%, about 96% to about 99.99%, about 97% to about 99.99%,
Approximately 98% to approximately 99.99%, approximately 99% to approximately 99.99%, approximately 99.9% to approximately 99.99%, approximately 95% to approximately 99.5%, approximately 96% to approximately 99
.5%, about 95% to about 99%, or about 97% to about 99% purity.

CXCR4 Antagonists Exemplary CXCR4 antagonists for use in conjunction with the compositions and methods described herein include formula (I)
Z-linker-Z' (I)
or a pharmaceutically acceptable salt thereof, where Z is:
(i) is a cyclic polyamine containing from 9 to 32 ring members, where 2 to 8 ring members are
are nitrogen atoms separated from each other by two or more carbon atoms; or
(ii) An amine represented by formula (IA)
where A comprises a monocyclic or bicyclic fused ring system containing at least one nitrogen atom, and B is H or a substituent consisting of 1 to 20 atoms;
and where Z' is:
(i) is a cyclic polyamine containing from 9 to 32 ring members, where 2 to 8 ring members are
are nitrogen atoms separated from each other by two or more carbon atoms;
(ii) An amine represented by formula (IB)
where A' comprises a monocyclic or bicyclic fused ring system containing at least one nitrogen atom, and B is H or a substituent consisting of 1 to 20 atoms; or
(iii) Substituent represented by formula (IC)
-N(R)-(CR ₂ ) _n -X (IC)
where each R is independently H or _C1 - _C6 alkyl, n is 1 or 2, and X is an aryl or heteroaryl group or a mercaptan;
wherein said linker is a bond, an optionally substituted alkylene (e.g., an optionally substituted C ₁ -C ₆ alkylene), an optionally substituted heteroalkylene (e.g., an optionally substituted optionally substituted alkenylene (e.g. optionally substituted _C2 - _C6 alkenylene) _, optionally substituted heteroalkenylene ( _e.g. , optionally substituted _C2 - _C6 heteroalkenylene),
optionally substituted alkynylene (e.g., optionally substituted _C2 - _C6 alkynylene), optionally substituted heteroalkynylene (e.g., optionally substituted
C ₂ -C ₆ heteroalkynylene), optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene, or optionally substituted heteroarylene It is.

In some embodiments, Z and Z' are each independently a cyclic polyamine containing 9 to 32 ring members, and 2 to 8 of the 9 to 32 ring members are It may be nitrogen atoms separated from each other by one or more carbon atoms. In some embodiments, Z and Z' are the same substituent. As an example, Z may be a cyclic polyamine containing 10 to 24 ring members. In some embodiments, Z can be a cyclic polyamine containing 14 ring members. In some embodiments, Z includes 4 nitrogen atoms. In some embodiments, Z is 1,4,8,11-tetraazocyclotetradecane.

In some embodiments, the linker is represented by the formula (ID),
where Ring D is an optionally substituted aryl group, an optionally substituted heteroaryl group, an optionally substituted cycloalkyl group, or an optionally substituted heterocycloalkyl group is; and
X and Y are each independently optionally substituted alkylene (e.g., optionally substituted C ₁ -C ₆ alkylene), optionally substituted heteroalkylene (e.g., optionally substituted optionally substituted C ₁ -C ₆ heteroalkylene), optionally substituted alkenylene (e.g., optionally substituted C ₂ -C ₆ alkenylene), optionally substituted heteroalkenylene ( for example, optionally substituted _C2 - _C6 heteroalkenylene),
optionally substituted alkynylene (e.g., optionally substituted C ₂ -C ₆ alkynylene), or optionally substituted heteroalkynylene (e.g., optionally substituted C ₂ - C ₆ heteroalkynylene).

For example, the linker may be represented by the formula (IE),
where Ring D is an optionally substituted aryl group, an optionally substituted heteroaryl group, an optionally substituted cycloalkyl group, or an optionally substituted heterocycloalkyl group is; and
X and Y are each independently optionally substituted alkylene (e.g., optionally substituted C ₁ -C ₆ alkylene), optionally substituted heteroalkylene (e.g., optionally substituted optionally substituted C ₁ -C ₆ heteroalkylene), optionally substituted alkenylene (e.g., optionally substituted C ₂ -C ₆ alkenylene), optionally substituted heteroalkenylene ( for example, optionally substituted _C2 - _C6 heteroalkenylene),
optionally substituted alkynylene (e.g., optionally substituted C ₂ -C ₆ alkynylene), or optionally substituted heteroalkynylene (e.g., optionally substituted C ₂ - C ₆ heteroalkynylene). In some embodiments, X and Y are each independently optionally substituted with _C1 - _C6 alkylene. In some embodiments, X and Y are the same substituents. In some embodiments, X and Y are each methylene,
It may be an ethylene, n-propylene, n-butylene, n-pentylene, or n-hexylene group. In some embodiments, X and Y are each methylene groups.

The linker is, for example, 1,3-phenylene, 2,6-pyridine, 3,5-pyridine, 2,5-thiophene, 4,4'-(2,2'-bipyrimidine), 2,9-(1 , 10-phenanthroline), etc. In some embodiments, the linker is 1,4-phenylene-bis-(methylene).

CXCR4 antagonists useful in conjunction with the compositions and methods described herein include formula (II), 1,1'-[1,4-phenylenebis(methylene)]-bis-1,4,8 , 11-tetra-azacyclotetradecane, plerixafor (herein referred to as "AMD3100" and "Mogivir"
), or a pharmaceutically acceptable salt thereof.

Additional CXCR4 antagonists that may be used in conjunction with the compositions and methods described herein include variants of plerixafor, such as the compounds described in U.S. Patent No. 5,583,131, which contain CXCR4 antagonists. is incorporated herein by reference as such. In some embodiments, the CXCR4 antagonist can be a compound selected from the group consisting of: 1,1′-[1,3-phenylenebis(methylene)]-
Bis-1,4,8,11-tetra-azacyclotetradecane; 1,1′-[1,4-phenylene-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane; Bis-zinc complex or bis-copper complex of 1,1′-[1,4-phenylene-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane; 1,1′-[ 3,3′-biphenylene-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane; 11,11′-[1,4-phenylene-bis-(methylene)]-bis- 1,4,7,11-tetraazacyclotetradecane; 1,11′-[1,4-phenylene-bis-(methylene)]-1,4,8,11-tetraazacyclotetradecane-1, 4,7 ,11-tetraazacyclotetradecane; 1,1′-[2,6-pyridine-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane; 1,1-[3,5 -pyridine-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane; 1,1′-[2,5-thiophene
-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane; 1,1′-[4,4′-(2,2
′-Bipyridine)-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane; 1,1
′-[2,9-(1,10-phenanthroline)-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane; 1,1′-[1,3-phenylene-bis -(methylene)]-bis-1,4,7,10-tetraazacyclotetradecane; 1,1′-[1,4-phenylene-bis-(methylene)]-bis-1,4,7,10- Tetraazacyclotetradecane; 1′-[5-nitro-1,3-phenylenebis(methylene)]bis-1,4,8,11-
Tetraazacyclotetradecane; 1′,1′-[2,4,5,6-tetrachloro-1,3-phenylenebis(
1,1′-[2,3,5,6-tetra-fluoro-1,4-phenylenebis(methylene)]bis-1, 4,8,11-tetraazacyclotetradecane; 1,
1′-[1,4-naphthylene-bis-(methylene)]bis-1,4,8,11-tetraazacyclotetradecane;
1,1′-[1,3-phenylenebis-(methylene)]bis-1,5,9-triazacyclododecane; 1,1′-[1
,4-phenylene-bis-(methylene)]-1,5,9-triazacyclododecane; 1,1′-[2,5-dimethyl
-1,4-phenylenebis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane; 1,1
′-[2,5-dichloro-1,4-phenylenebis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane; 1,1′-[2-bromo-1,4- Phenylenebis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane; and 1,1′-[6-phenyl-2,4-pyridinebis-(methylene)]-bis
-1,4,8,11-tetraazacyclotetradecane.

In some embodiments, the CXCR4 antagonist is a compound described in US 2006/0035829, the disclosure of which is incorporated herein by reference as it relates to CXCR4 antagonists. In some embodiments, the CXCR4 antagonist may be a compound selected from the group consisting of: 3,7,11,17-tetraazabicyclo(13.3.1)heptadeca-1(17), 13,15-triene; 4,7,10,17-tetraazabicyclo(13.3.1)
Heptadeca-1(17),13,15-triene; 1,4,7,10-tetraazacyclotetradecane; 1,4,7-triazacyclotetradecane; and 4,7,10-triazabicyclo (13.3. 1) Heptadeca-1(17),13
,15-triene.

Said CXCR4 antagonist may be a compound described in WO 2001/044229, the disclosure of which is incorporated herein by reference as it relates to CXCR4 antagonists. In some embodiments, the CXCR4 antagonist can be a compound selected from the group consisting of: N-[4-(11-fluoro-1,4,7-triazacyclotetradecanyl) -1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;
N-[4-(11,11-difluoro-1,4,7-triazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine; N-[4- (1,4,7-triazacyclotetradecan-2-onyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine; N-[12-(5-oxa-1,9-
diazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine; N-[4-(11-oxa-1,4,7-triazacyclotetradecanyl) -1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine; N-[4-(11-thia-1,4,7-triazacyclotetradecanyl)-1,4-phenylenebis (methylene)]-2-(aminomethyl)pyridine; N-[4-(11-sulfoxo-1,4,7-triazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2- (aminomethyl)pyridine; N-[4-(11-sulfono-1,4,7-triazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine; and N-[4-(3-carboxo-1,4,7-triazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine.

Additional CXCR4 antagonists useful in conjunction with the compositions and methods described herein include compounds described in WO 2000/002870, the disclosure of which, as related to CXCR4 antagonists, includes: Incorporated herein by reference. In some embodiments, the CXCR4 antagonist can be a compound selected from the group consisting of: N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylene Bis-(methylene)]-2-(aminomethyl)pyridine; N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4
-phenylenebis(methylene)]-N-methyl-2-(aminomethyl)pyridine; N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-4 -(aminomethyl)pyridine
; N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-3-(
N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-(2-aminomethyl-5-methyl)pyridine; N-[1 ,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-2-(aminoethyl)pyridine; N-[1,
4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-2-(aminomethyl)thiophene; N-[1,4,8,11-tetraazacyclotetra-decanyl- 1,4-phenylenebis(
methylene)]-2-(aminomethyl)mercaptan; N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-2-amino benzylamine; 1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-4-amino benzylamine; N
-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-4-(aminoethyl)imidazole; N-[1,4,8,11-tetraazacyclo Tetra-decanyl-1,4-phenylenebis(methylene)]-benzylamine; N-[4-(1,4,7-triazacyclotetra-decanyl)-1,4
-phenylenebis(methylene)]-2-(aminomethyl)pyridine; N-[7-(4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl) )-1,4-phenylenebis(methylene)]
-2-(aminomethyl)pyridine; N-[7-(4,7,10-triazabicyclo[13.3.1]heptadeca-1(17)
,13,15-trienyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine; N-[1-
(1,4,7-triazacyclotetra-decanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine; N-[4-[4,7,10,17-tetraaza Bicyclo[13.3.1]heptadeca-1(17),13,15-
trienyl]-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine; N-[4-[4,7,10
-triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(
methylene)]-2-(aminomethyl)pyridine; N-[1,4,8,11-tetraazacyclotetradecanyl-
1,4-phenylenebis(methylene)]-purine; 1-[1,4,8,11-tetraazacyclotetradecanyl
-1,4-phenylenebis(methylene)]-4-phenylpiperazine; N-[4-(1,7-diazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(amino methyl)pyridine; and N-[7-(4,
10-Diazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl)-1,4-phenylenebis(
methylene)]-2-(aminomethyl)pyridine.

In some embodiments, the CXCR4 antagonist is a compound selected from the group consisting of: 1-[2,6-dimethoxypyrid-4-yl(methylene)]-1,4,8,11 -Tetraazacyclotetradecane; 1-[2-chloropyrid-4-yl(methylene)]-1,4,8,11-tetraazacyclotetradecane; 1-[2,6-dimethylpyrid-4-yl(methylene)] -1,4,8,11-tetraazacyclotetradecane; 1-[2-methylpyrid-4-yl(methylene)]-1,4,8,11-tetraazacyclotetradecane; 1-[2,6-dichloropyrido -4-yl(methylene)]-1,4,8,11-tetraazacyclotetradecane; 1-[2-chloropyrid-5-yl(methylene)]-1,4,8,11-tetraazacyclotetradecane; and 7-[4-methylphenyl (methylene)]-4,7,10,17-tetraazabicyclo[13.3.1]
Heptadeca-1(17),13,15-triene.

In some embodiments, the CXCR4 antagonist is a compound described in U.S. Patent No. 5,698,546, the disclosure of which relates to CXCR4 antagonists.
Incorporated herein by reference. In some embodiments, the CXCR4 antagonist can be a compound selected from the group consisting of: 7,7′-[1,4-phenylene-
Bis(methylene)]bis-3,7,11,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-
Triene; 7,7′-[1,4-phenylene-bis(methylene)]bis[15-chloro-3,7,11,17-tetraazabicyclo[13.3.1]heptadeca-1 (17),13, 15-triene]; 7,7′-[1,4-phenylene-bis
(methylene)]bis[15-methoxy-3,7,11,17-tetraazabicyclo[13.3.1]heptadeca-1(17)
,13,15-triene]; 7,7′-[1,4-phenylene-bis(methylene)]bis-3,7,11,17-tetraazabicyclo[13.3.1]-heptadeca-13,16- Trien-15-one; 7,7′-[1,4-phenylene-bis(
8,8′-[1,4-phenylene-bis(methylene)]bis-4,7,10,17-tetraazabicyclo[13.3.1]-heptadeca-1(17),13,15-triene; )]bis-4,8,12,19-tetraazabicyclo[15.
3.1]nonadeca-1(19),15,17-triene; 6,6′-[1,4-phenylene-bis(methylene)]bis-3,6
,9,15-tetraazabicyclo[11.3.1]pentadeca-1 (15),11,13-triene; 6,6′-[1,3-phenylene-bis(methylene)]bis-3,6,9 ,15-tetraazabicyclo[11.3.1]pentadeca-1 (15
),11,13-triene; and 17,17′-[1,4-phenylene-bis(methylene)]bis-3,6, 14,17,23
,24-Hexaazatricyclo[17.3.1.1 ^8,12 ]tetracosa-1(23),8,10,12(24),19,21-hexaene.

In some embodiments, the CXCR4 antagonist is a compound described in U.S. Patent No. 5,021,409, the disclosure of which relates to CXCR4 antagonists.
Incorporated herein by reference. In some embodiments, the CXCR4 antagonist can be a compound selected from the group consisting of: 2,2′-bicyclam, 6,
6′-bicyclam; 3,3′-(bis-1,5,9,13-tetraazacyclohexadecane); 3,3′-(bis-1,5,8,11,14-pentaazacyclohexadecane); Methylene (or polymethylene)
Di-1-N-1,4,8,11-tetraaza cyclotetradecane; 3,3′-bis-1,5,9,13-tetraazacyclohexadecane; 3,3′-bis-1,5,8 ,11,14-pentaazacyclohexadecane; 5,5′-bis-1,4,8,11-tetraazacyclotetradecane; 2,5′-bis-1,4,8,11-tetraazacyclotetradecane; 2,6′-bis-1,4,8,11-tetraazacyclotetradecane; 11,11′-(1,2-ethanediyl)bis-1,4,8,11-tetraazacyclotetradecane; 11,11 ′-(1,2-propanediyl
)bis-1,4,8,11-tetraazacyclotetradecane; 11,11′-(1,2-butanediyl)bis-1,4,
8,11-tetraazacyclotetradecane; 11,11′-(1,2-pentanediyl)bis-1,4,8,11-tetraazacyclotetradecane; and 11,11′-(1,2-hexanediyl) Bis-1,4,8,11-tetraazacyclotetradecane.

In some embodiments, the CXCR4 antagonist is a compound described in WO 2000/056729, the disclosure of which is incorporated herein by reference as it relates to CXCR4 antagonists. In some embodiments, the CXCR4 antagonist is
It may be a compound selected from the group consisting of: N-(2-pyridinylmethyl)-N′-(
N-(2-pyridinylmethyl)-N′-(5,6,7, 8-Tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(6,7-dihydro-5H-cyclopenta[b]pyridin-7-yl)-1 ,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(1,2,3,4-tetrahydro-1-naphthalenyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl )-N
′-(1-naphthalenyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(8-quinolinyl)-1,4-benzenedimethaneamine; N-(2-pyridinylmethyl) -N′-[2-[(2-pyridinylmethyl)amino]ethyl]-N′-(1-methyl-1,2,3,4-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-[(1H-imidazol-2-ylmethyl)amino]ethyl]-N′-(1-methyl-1,2,3,4-tetrahydro-8-quinolinyl )-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(1,2,3,4-tetrahydro-8-quinolinyl)-1
,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-[(1H-imidazol-2-ylmethyl)amino]ethyl]-N′-(1,2,3,4-tetrahydro -1-naphthalenyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(2-phenyl-5,6,7,8-tetrahydro-8-quinolinyl)-1,4- Benzenedimethanamine; N,N′-bis(2-pyridinylmethyl)-N′-(2-phenyl-5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;

N-(2-pyridinylmethyl)-N′-(5,6,7,8-tetrahydro-5-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(1H- imidazol-2-ylmethyl)-N′-(5,6
,7,8-tetrahydro-5-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(1H-imidazol-2-ylmethyl)-N′-(5,6, 7,8-tetrahydro-8-quinolinyl)-1
,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[(2-amino-3-phenyl)propyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl) -1,4-benzenedimethanamine; N-(2-
pyridinylmethyl)-N′-(1H-imidazol-4-ylmethyl)-N′-(5,6,7,8-tetrahydro-8
-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(2-quinolinylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl) -1,4-benzenedimethanamine;
N-(2-pyridinylmethyl)-N′-(2-(2-naphthoyl)aminoethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine ; N-(2-pyridinylmethyl)-N′-[(S)-(
2-acetylamino-3-phenyl)propyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,
4-Benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[(S)-(2-acetylamino-3-phenyl)propyl]-N′-(5,6,7,8-tetrahydro- 8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[3-((2-naphthalenylmethyl)amino)propyl]-N′-
(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-(S)-pyrrolidinylmethyl]-N′ -(5,6,7,8-tetrahydro-8-quinolinyl)
-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-(R)-pyrrolidinylmethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl )-1,4-benzenedimethanamine; N-(2-
N-(2-pyridinylmethyl)- N′-[2-pyrrolylmethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-
N-(2-pyridinylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N′-[2-thiazolylmethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2
-pyridinylmethyl)-N′-[2-furanylmethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl
)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-[(phenylmethyl)amino]ethyl]-N′-(5,6,7,8-tetrahydro-8 -quinolinyl)-1,4-benzenedimethanamine;
N-(2-pyridinylmethyl)-N′-(2-aminoethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2 -pyridinylmethyl)-N′-3-pyrrolidinyl-N′-
(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-4-piperidinyl-N′-(5,6,7, 8-Tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-[(phenyl)amino]ethyl]-N′-(5,6
,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(7-methoxy-1,2,3,4-tetrahydro-2-naphthalenyl) )-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(6-methoxy-1,2,3,4-tetrahydro-2-naphthalenyl
)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(1-methyl-1,2,3,4-tetrahydro-2-naphthalenyl)-1,4-benzenedimethanamine ; N-(2-pyridinylmethyl)-N′-
(7-methoxy-3,4-dihydronaphthalenyl)-1-(aminomethyl)-4-benzamide; N-(2-pyridinylmethyl)-N′-(6-methoxy-3,4-dihydronaphthalenyl )-1-(aminomethyl)-4-benzamide; N-(2-pyridinylmethyl)-N′-(1H-imidazol-2-ylmethyl)-N′-(7-methoxy-1,2,3,4- N-(2-pyridinylmethyl)-N′-(8-hydroxy-1,2,3,4-tetrahydro-2-naphthalenyl)-1,4 -Benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(1H-imidazol-2-ylmethyl)-N′-
(8-hydroxy-1,2,3,4-tetrahydro-2-naphthalenyl)-1,4-benzenedimethanamine; N
-(2-pyridinylmethyl)-N′-(8-fluoro-1,2,3,4-tetrahydro-2-naphthalenyl)-1,4-
Benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(1H-imidazol-2-ylmethyl
)-N′-(8-fluoro-1,2,3,4-tetrahydro-2-naphthalenyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(5,6, 7,8-tetrahydro-7-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(1H-imidazol-2-ylmethyl)-N′-(5
,6,7,8-tetrahydro-7-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-[(2-naphthalenylmethyl)amino]ethyl] -N′-(5,6,7,8-tetrahydro-8-
quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-(isobutylamino)ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl )-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-[(2-pyridinylmethyl)amino]ethyl]-N′-(5,6
,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-[(2-furanylmethyl)amino]ethyl]-N′-( 5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(2-guanidinoethyl
)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-[bis-[(2 -methoxy)phenylmethyl]amino]ethyl]-N′-(5,6,7
,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)
-N′-[2-[(1H-imidazol-4-ylmethyl)amino]ethyl]-N′-(5,6,7,8-tetrahydro-
8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-[(1H-imidazol-2-ylmethyl)amino]ethyl]-N′-(5,6 ,7,8-tetrahydro-8-quinolinyl)-1
,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-(phenylureido)ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1, 4-Benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[[N″-(n-butyl)carboxamido]methyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl )-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(carboxamidomethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4- Benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[(N″-phenyl)carboxamidomethyl]-N′-(5,6,7
,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)
-N′-(carboxymethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(phenyl N-(2-pyridinylmethyl)-N′-(1H-benzimidazole-2- N-(2-pyridinylmethyl)-N′-(5,6-dimethyl-1H -benzimidazole
-2-ylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine (hydrobromide salt); N-(2-pyridinylmethyl)-N′- (5-nitro-1H-benzimidazol-2-ylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl) -N′-[(1H)-5-azabenzimidazol-2-ylmethyl]-
N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N-(4-phenyl-1H-imidazol-2-ylmethyl )-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-[2-(2-pyridinyl)ethyl ]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(2-benzoxazolyl)- N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(trans-2-aminocyclohexyl)-N′ -(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(2-phenylethyl)-N′-(5, 6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(3
-phenylpropyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N′-(trans-2-amino cyclopentyl)-N′-(5,6,7,
8-Tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8 -tetrahydro-8-quinolinyl)-glycinamide; N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-
Tetrahydro-8-quinolinyl)-(L)-alaninamide; N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8 -quinolinyl)-(L)-aspartamide; N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl )-pyrazinamide; N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl)-(L)-proline Amide; N-
[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro
-8-quinolinyl)-(L)-lysinamide; N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl )-Benzamide; N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl
)-Picolinamide; N′-benzyl-N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl)- Urea; N′-phenyl-N-[[4-[[(2
-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl)-urea; N-(6,7,8,9-tetrahydro-5H-cyclohepta[bacterium Pyridin-9-yl)-4-[[(2-pyridinylmethyl)amino]methyl]benzamide; N-(5,6,7,8-tetrahydro-
8-quinolinyl)-4-[[(2-pyridinylmethyl)amino]methyl]benzamide; N,N′-bis(2-
N,N′-bis(2-pyridinylmethyl)-N′-(6,7 ,8,9-tetrahydro-5H-cyclohepta[
Bacterial pyridin-9-yl)-1,4-benzenedimethanamine; N,N′-bis(2-pyridinylmethyl)-N′-(6,7-dihydro-5H-cyclopenta[bacterial pyridin-7-yl) -1,4-benzenedimethanamine; N,N′-bis(2-pyridinylmethyl)-N′-(1,2,3,4-tetrahydro-1-naphthalenyl)-1,4-benzenedimethanamine; N,N′-bis(2-pyridinylmethyl)-N′-[(5,6,7
,8-tetrahydro-8-quinolinyl)methyl]-1,4-benzenedimethanamine; N,N′-bis(2-pyridinylmethyl)-N′[(6,7-dihydro-5H-cyclopenta[bacteriapyridine- 7-yl)methyl]-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N-(2-methoxyethyl)-N′-
(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(2-pyridinylmethyl)-N-[2-(4-methoxyphenyl)ethyl]-N′- (5,6,7,8-tetrahydro-8-quinolinyl)
-1,4-benzenedimethanamine; N,N′-bis(2-pyridinylmethyl)-1,4-(5,6,7,8-tetrahydro-8-quinolinyl)benzenedimethanamine; N-[( 2,3-dimethoxyphenyl)methyl]-N
′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N,N′-bis(2-pyridinylmethyl)-N-[ 1-(N″-phenyl-N″-methylureido)-
4-piperidinyl]-1,3-benzenedimethanamine; N,N′-bis(2-pyridinylmethyl)-N-[N
″-p-Toluenesulfonylphenylalanyl)-4-piperidinyl]-1,3-benzenedimethanamine; N,N′-bis(2-pyridinylmethyl)-N-[1-[3-(2-chlorophenyl) -5-methyl-isoxazol-4-oyl]-4-piperidinyl]-1,3-benzenedimethanamine;
N-[(2-hydroxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[bacterial pyridin-9-yl)-1,4- Benzenedimethanamine; N-[(4-
cyanophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[bacterial pyridin-9-yl)-1,4-benzenedimethanamine; N -[(4-cyanophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-
Benzenedimethanamine; N-[(4-acetamidophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedi Methanamine; N-[(4-phenoxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[bacterial pyridin-9-yl)-1 ,4-benzenedimethanamine; N-[(1-methyl-2-
carboxamido)ethyl]-N,N′-bis(2-pyridinylmethyl)-1,3-benzenedimethanamine
; N-[(4-benzyloxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[bacterial pyridin-9-yl)-1, 4-benzenedimethanamine;
N-[(thiophen-2-yl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5
H-cyclohepta[bacterial pyridin-9-yl)-1,4-benzenedimethanamine; N-[1-(benzyl)-3-pyrrolidinyl]-N,N′-bis(2-pyridinylmethyl)-1,3 -benzenedimethanamine;
N-[[1-methyl-3-(pyrazole-3-yl)]propyl]-N,N′-bis(2-pyridinylmethyl)-1,3-
Benzenedimethanamine; N-[1-(phenyl)ethyl]-N,N′-bis(2-pyridinylmethyl)-1,
3-benzenedimethanamine; N-[(3,4-methylenedioxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-diclohepta[b ]pyridin-9-yl)-1,4-benzenedimethanamine; N-[1-benzyl-3-carboxymethyl-4-piperidinyl]-N,N′-bis(
2-pyridinylmethyl)-1,3-benzenedimethanamine; N-[(3,4-methylenedioxyphenyl
)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(3-pyridinylmethyl)-N′ -(2-pyridinylmethyl)-N-(6,7,8,9-
Tetrahydro-5H-diclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine; N-[[
1-Methyl-2-(2-tolyl)carboxamido]ethyl]-N,N′-bis(2-pyridinylmethyl)-1,3-
Benzenedimethanamine; N-[(1,5-dimethyl-2-phenyl-3-pyrazolinon-4-yl)methyl
]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-[(4-propoxyphenyl)methyl]- N′-(2-pyridinylmethyl)-N-(6,7,8
,9-tetrahydro-5H-diclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine; N
-(1-phenyl-3,5-dimethylpyrazolin-4-ylmethyl)-N′-(2-pyridinylmethyl)-N-(5,
6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-[H-imidazole-4
-ylmethyl]-N,N′-bis(2-pyridinylmethyl)-1,3-benzenedimethanamine; N-[(3-methoxy-4,5-methylenedioxyphenyl)methyl]-N′-(2 -pyridinylmethyl)-N-(6,7,8,9-
Tetrahydro-5H-diclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;
3-cyanophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-diclohepta[b]pyridin-9-yl)-1,4-benzenedimethane Amine; N-[(3-cyanophenyl)
methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; )-N′-(2-pyridinylmethyl)
-N-(6,7,8,9-tetrahydro-5H-diclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine; N-(5-ethylthiophen-2-ylmethyl)-N ′-(2-pyridinylmethyl)-N-(5,6,7,8-
N-[(2,6-difluorophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9- Tetrahydro-5H-dichlorohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine; N-[(2,6-difluorophenyl)methyl]-N
′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-[(2-difluoromethoxyphenyl)methyl]-N′ -(2-pyridinylmethyl)-N-(6
,7,8,9-tetrahydro-5H-diclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine; N-(2-difluoromethoxyphenylmethyl)-N′-(2-pyridinylmethyl) -N-(5,6,7,8-
Tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(1,4-benzodioxane-6
-ylmethyl)-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-diclohepta[b]
Pyridin-9-yl)-1,4-benzenedimethanamine; N,N′-bis(2-pyridinylmethyl)-N-[1
-(N″-phenyl-N″-methylureido)-4-piperidinyl]-1,4-benzenedimethanamine; N
,N′-bis(2-pyridinylmethyl)-N-[N″-p-toluenesulfonylphenylalanyl)-4-piperidinyl]-1,4-benzenedimethanamine; N-[1-(3-pyridinecarboxamide) )-4-piperidinyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine; N-[1-(cyclopropylcarboxamide)-4-piperidinyl]-N,N′-bis (2-pyridinylmethyl)-1,4-benzenedimethanamine; N-[1-(1-phenylcyclopropylcarboxamide)-4-piperidinyl
]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine; N-(1,4-benzodioxan-6-ylmethyl)-N′-(2-pyridinylmethyl)-N-( 5,6,7,8-tetrahydro-8-quinolinyl
)-1,4-benzenedimethanamine; N-[1-[3-(2-chlorophenyl)-5-methyl-isoxazole-4-carboxamido]-4-piperidinyl]-N,N′-bis(2 -pyridinylmethyl)-1,4-benzenedimethanamine; N-[1-(2-thiomethylpyridine-3-carboxamide)-4-piperidinyl]-N,N
′-Bis(2-pyridinylmethyl)-1,4-benzenedimethanamine; N-[(2,4-difluorophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7, 8-Tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(1-methylpyrrol-2-ylmethyl)-N′-(2-pyridinylmethyl)
-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-[(2-hydroxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N -(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-[(3-methoxy-4,5-methylenedioxyphenyl)methyl]-N′-( N-(3-pyridinylmethyl)-N′-(2-pyridinylmethyl)-N -(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-[2-(N″-morpholinomethyl)
-1-cyclopentyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine; N-
[(1-methyl-3-piperidinyl)propyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine; N-(1-methylbenzimidazol-2-ylmethyl)-N′ -(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-[1-(benzyl)-3-pyrrolidinyl]-N ,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine; N-[[(1-phenyl-3-(N″-morpholino)]propyl]-N,N′-bis(2- pyridinylmethyl
)-1,4-benzenedimethanamine; N-[1-(iso-propyl)-4-piperidinyl]-N,N′-bis(2-
pyridinylmethyl)-1,4-benzenedimethanamine; N-[1-(ethoxycarbonyl)-4-piperidinyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8 -quinolinyl)-1,4-benzenedimethanamine; N-[(1-methyl-3-pyrazolyl)propyl]-N′-(2-pyridinylmethyl)
-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-[1-methyl-2
-(N″,N″-diethylcarboxamido)ethyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine; N-[(1-methyl-2-phenylsulfonyl)ethyl] -N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-[(2-
Chloro-4,5-methylenedioxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-
N-[1-methyl-2-[N″-(4-chlorophenyl)carboxamido]ethyl]-N′-(2-pyridinylmethyl)-N- (5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N-(1-acetoxyindol-3-ylmethyl)-N′-(2-pyridinylmethyl)-N-( 6,7,8,9-tetrahydro-5H-diclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine; N-[(3-benzyloxy-4-methoxyphenyl)
)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-diclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine; N- (3-quinolylmethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine; N
-[(8-hydroxy)-2-quinolylmethyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-diclohepta[b]pyridin-9-yl)- 1,4-Benzenedimethanamine; N-(2-quinolylmethyl)-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-diclohepta[b]pyridine-9 -yl)-1,4-benzenedimethanamine; N-[(4-acetamidophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-diclohepta [b]Pyridin-9-yl)-1,4-benzenedimethane; N-[1H-imidazol-2-ylmethyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethane Amine; N-(3-quinolylmethyl)-N′-
(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-diclohepta[b]pyridin-9-yl)-
1,4-benzenedimethanamine; N-(2-thiazolylmethyl)-N′-(2-pyridinylmethyl)-
N-(6,7,8,9-tetrahydro-5H-diclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine; N-(4-pyridinylmethyl)-N′-(2-pyridinylmethyl )-N-(6,7,8,9-tetrahydro-
5H-diclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine; N-[(5-benzyloxy)benzo[b]pyrrol-3-ylmethyl]-N,N′-bis(2 -pyridinylmethyl)-1,4-benzenedimethanamine; N-(1-methylpyrazol-2-ylmethyl)-N′-(2-pyridinylmethyl)-N-(
6,7,8,9-tetrahydro-5H-diclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine; N-[(4-methyl)-1H-imidazol-5-ylmethyl]- N,N′-bis(2-pyridinylmethyl)-1
,4-benzenedimethanamine; N-[[(4-dimethylamino)-1-naphthalenyl]methyl]-N,N′-
Bis(2-pyridinylmethyl)-1,4-benzenedimethanamine; N-[1,5-dimethyl-2-phenyl-
3-pyrazolinon-4-ylmethyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine; N-[1-[(1-acetyl-2-(R)-prolinyl]- 4-Piperidinyl]-N-[2-(2-pyridinyl)
ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine; N-[1-[2-acetamidobenzoyl-4-piperidinyl]-4-piperidinyl]-N-[2-(2- pyridinyl)ethyl]-N′-(2-
Pyridinylmethyl)-1,3-benzenedimethanamine; N-[(2-cyano-2-phenyl)ethyl]-N
′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-diclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine; N-[(N″- Acetyltryptophanyl)-4-piperidinyl
]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine
; N-[(N″-benzoylvalinyl)-4-piperidinyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-
N-[(4-dimethylaminophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-diclohepta)-1,3-benzenedimethanamine; [b]Pyridine-
9-yl)-1,4-benzenedimethanamine; N-(4-pyridinylmethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1 ,4-benzenedimethanamine; N-(1-methylbenzimidazol-2-ylmethyl)-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-diclohepta[b] Pyridin-9-yl)-1,4-benzenedimethanamine; N-[1-butyl-
4-Piperidinyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine; [2-(2-pyridinyl)ethyl]-N′-(
2-pyridinylmethyl)-1,3-benzenedimethanamine

;
N-[1-(benzyl)-3-pyrrolidinyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine; N-[(1 -methyl)benzo[b]pyrrol-3-ylmethyl]-N
-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine; N
-[1H-imidazol-4-ylmethyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine; N-[1-(benzyl) -4-piperidinyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,4-benzenedimethanamine; N-[1-methylbenzimidazol-2-ylmethyl] -N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)
-1,4-benzenedimethanamine; N-[(2-phenyl)benzo[b]pyrrol-3-ylmethyl]-N-[2
-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,4-benzenedimethanamine; N-[(
6-Methylpyridin-2-yl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro
-8-quinolinyl)-1,4-benzenedimethanamine; N-(3-methyl-1H-pyrazol-5-ylmethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8 -tetrahydro-8-quinolinyl)-1,3-benzenedimethanamine; N-[(2-methoxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7
,8-tetrahydro-8-quinolinyl)-1,3-benzenedimethanamine; N-[(2-ethoxyphenyl
)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-diclohepta[b]pyridin-9-yl)-1,3-benzenedimethanamine; N- (Benzyloxyethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,3-benzenedimethanamine; N-
[(2-ethoxy-1-naphthalenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,3-benzenedimethanamine; N -[(6-methylpyridin-2-yl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,3-benzenedimethanamine ; 1-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]guanidine; N-(2-pyridinylmethyl)-N-(8-methyl-8-azabicyclo[3.2.1]octane-3 -il)-1
,4-benzenedimethanamine; 1-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]homopiperazine; 1-[[3-[[(2-pyridinylmethyl)amino]methyl]phenyl ]methyl]
homopiperazine; trans and cis-1-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-3,5-piperidinediamine; N,N′-[1,4-phenylenebis(methylene )]Bis-4-
(2-pyrimidyl)piperazine; 1-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-1-(2-pyridinyl)methylamine; 2-(2-pyridinyl)-5-[ [(2-pyridinylmethyl)amino]methyl]-1,2,3,4-tetrahydroisoquinoline; 1-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-3,4-diaminopyrrolidine ; 1-[[4-[[(2-pyridinylmethyl)
8-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-2,5,8-triaza-3- Oxabicyclo [4.3.0
]nonane; and 8-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-2,5,8-
triazabicyclo[4.3.0]nonane.

Additional CXCR4 antagonists that may be used in conjunction with the compositions and methods described herein include WO 2001/085196, WO 1999/050461, WO 2001/094420, and WO 2003/0905.
12, the disclosures of each of which are incorporated herein by reference as they relate to compounds that inhibit CXCR4 activity or expression.

Expansion of Hematopoietic Stem and Progenitor Cells Prior to infusion into a patient, hematopoietic and progenitor cells may be expanded ex vivo, eg, by contacting the cells with an antagonist of the aryl hydrocarbon receptor.
Aryl hydrocarbon receptor antagonists useful in conjunction with the compositions and methods described herein include those described in U.S. Pat. No. 9,580,426, the disclosure of which includes: Incorporated herein by reference in its entirety.

In some embodiments, the aryl hydrocarbon receptor antagonist comprises the formula
aryl hydrocarbon receptor antagonist represented by (III),
here:
L is -NR _5a (CH ₂ ) _2-3 , -NR _5a (CH ₂ ) ₂ NR _5b -, -NR _5a (CH ₂ ) ₂ S-, -NR _5a CH ₂ CH(OH)- and NR _5a CH(C
_H3 ) _CH2- ; where _R5a and _R5b are independently selected from hydrogen and _C1-4 alkyl;
R ₁ is selected from thiophenyl, 1H-benzimidazolinyl, isoquinolinyl, 1H-imidazopyridinyl, benzothiophenyl, pyrimidinyl, pyridinyl, pyrazinyl, pyridazinyl, and thiazolyl; , said thiophenyl, 1H
- R ₁ of benzimidazolinyl, isoquinolinyl, 1H-imidazopyridinyl, benzothiophenyl, pyrimidinyl, pyridinyl, pyrazinyl, pyridazinyl, or thiazolyl, cyano, hydroxy, C _1-4 alkyl, C _1-4 alkoxy, Halo, halo-substituted -C _1-4 alkyl, halo-substituted -C _1-4 alkoxy, amino, -C(O)R _8a , -S(O) _0-2 R _8a , -C(O)OR _8a , and -C(O)NR _8a R _8b
optionally substituted with 1 to 3 radicals independently selected from; where R _8a and R _8b are independently selected from hydrogen and C _1-4 alkyl;
R ₂ is -S(O) ₂ NR _6a R _6b , -NR _6a C(O)R _6b -, -NR _6a C(O)R _6b R _6c , phenyl, 1H-pyrrolopyridin-3-yl, 1H -pyrrolopyridin-5-yl, 1H-indolyl thiophenyl, pyridinyl, 1H
-1,2,4-triazolyl, 2-oxoimidazolidinyl, 1H-pyrazolyl, 2-oxo-2,3-dihydro-1H-benzimidazolyl and 1H-indazolyl; where R _6a , R _6b and R _6c are independently selected from hydrogen and C _1-4 alkyl; and said phenyl, 1H-pyrrolopyridin-3-yl, 1H-pyrrolo[2,3-b]pyridin-5-yl, 1H -indolyl, thiophenyl, pyridinyl, 1H-1,2,4-triazolyl, 2-oxoimidazolidinyl, 1H-pyrazolyl, 2-
R ₂ of oxo-2,3-dihydro-1H-benzimidazolyl or 1H-indazolyl is hydroxy,
Halo, methyl, methoxy, amino, -O(CH ₂ ) ₂ NR _7a R _7b , -S(O) ₂ NR _7a R _7b , -OS(O) ₂ NR _7a R _7b ,
and -NR _7a optionally substituted with 1 to 3 radicals independently selected from S(O) ₂ R _7b ; where R _7a and R _7b are independently selected from hydrogen and C _1-4 alkyl. selected by;
R ₃ is selected from hydrogen, C _1-4 alkyl and biphenyl; and
R ₄ is C _1-10 alkyl, prop-1-en-2-yl, cyclohexyl, cyclopropyl, 2
-(2-oxopyrrolidin-1-yl)ethyl, oxetan-2-yl, oxetan-3-yl, benzhydryl, tetrahydro-2H-pyran-2-yl, tetrahydro-2H-pyran-3-yl, phenyl, tetrahydrofuran -3-yl, and benzyl, (4-pentylphenyl)(phenyl)methyl and 1-(1-(2-oxo-6,9,12-trioxa-3-azatetradecan-14-yl)-1H-1 ,2,3-triazol-4-yl)ethyl, wherein said alkyl is selected from cyclopropyl, cyclohexyl, 2-(2-oxopyrrolidin-1-yl)ethyl, oxetan-3-yl, oxetane- 2-
yl, benzhydryl, tetrahydro-2H-pyran-2-yl, tetrahydro-2H-pyran-3-yl, phenyl, tetrahydrofuran-3-yl, benzyl, (4-pentylphenyl)(phenyl
)methyl or 1-(1-(2-oxo-6,9,12-trioxa-3-azatetradecan-14-yl)-1H-1,2,3
-triazol-4-yl)ethyl optionally substituted with 1 to 3 radicals independently selected from hydroxy, C _1-4 alkyl and halo-substituted -C _1-4 alkyl; or salts thereof; There are things to do.

In some embodiments, aryl hydrocarbon receptor antagonists useful in conjunction with the compositions and methods described herein include SR-1 of formula (1) below.

Methods for Recombinant Expression of Peptides and Proteins The peptides and proteins described herein (e.g., Gro-β, Gro-β T, Gro-β N6
9D, Gro-β T N65D, and variants thereof) may be expressed in a host cell, for example, by delivering a nucleic acid encoding the corresponding peptide or protein into the host cell. The following sections describe various techniques that can be used to introduce nucleic acids encoding the peptides and proteins described herein into host cells for the purpose of recombinant expression.

transfection technology
CX such as Gro-β, Gro-β T, Gro-β N69D, Gro-β T N65D, or variants thereof
Techniques that can be used to introduce polynucleotides, such as nucleic acids encoding CR2 agonists, into cells (eg, mammalian cells, eg, human cells) are known to those skilled in the art. In some embodiments, electroporation can be used to permeabilize mammalian cells (eg, human cells) by applying an electrostatic potential to the cells of interest. Mammalian cells, such as human cells, exposed to an external electric field in this manner are then pretreated to incorporate foreign nucleic acids. Electroporation of mammalian cells, e.g.
Chu et al. (1987) Nucleic Acids Research 15:1311, the disclosure of which is incorporated herein by reference. A similar technique, nucleofection (Nuc)
leofection ( ^TM ) utilizes an applied electric field to stimulate the uptake of foreign polynucleotides into the nucleus of eukaryotic cells. Nucleofection ^TM and protocols useful for implementing this technique are described, for example, in Distler et al. (2005) Experimental Dermatology 14:315, and in the US
2010/0317114, the disclosures of each of which are incorporated herein by reference.

Additional techniques useful for transfecting host cells for the purpose of expressing recombinant peptides and proteins include the squeeze-poration method. This technique rapidly mechanically deforms cells to stimulate the uptake of foreign DNA through membrane pores that form in response to the applied stress. This technology is advantageous in that it does not require vectors to deliver nucleic acids to cells such as human cells. Squeeze-poration is described, for example, by Sharei et al. (2013) Journal of Visualized Expe.
riments 81:e50980, the disclosure of which is incorporated herein by reference.

Lipofection is another technique useful for transfecting cells. This method involves loading nucleic acids into liposomes that often display cationic functional groups, such as quaternary or protonated amines, toward the outside of the liposome. including. This promotes electrostatic interactions between the liposomes and the cells due to the anionic nature of the cell membranes, ultimately leading to direct fusion of the liposomes and the cell membranes, for example. or endocytosis of the complex results in the uptake of foreign nucleic acids. Lipofection is described in detail in, for example, US Pat. No. 7,442,386, the disclosure of which is incorporated herein by reference. Similar techniques that utilize ionic interactions with the cell membrane to induce uptake of foreign nucleic acids include contacting cells with cationic polymer-nucleic acid complexes. Exemplary cationic molecules associated with polynucleotides that impart a positive charge suitable for interaction with the cell membrane include activated dendrimers (e.g., Dennig (2003) Topics
in Current Chemistry 228:227, the disclosure of which is incorporated herein by reference), and diethylaminoethyl (DEAE)-dextran, whose use as a transfection reagent has been described, for example, by Gulick et al. 1997)Current Protocols in Molec
ular Biology 40:I:9.2:9.2.1, the disclosure of which is incorporated herein by reference). Magnetic beads are another tool that can be used to gently and efficiently transfect cells, as this method utilizes an applied magnetic field to uptake nucleic acids. This technique is described in detail in, for example, US 2010/0227406, the disclosure of which is incorporated herein by reference.

Another useful tool for inducing the uptake of foreign nucleic acids by cells is laserfection. This technique involves exposing the cells to specific wavelengths of electromagnetic radiation to gently permeabilize the cells and allow the polynucleotide to penetrate the cell membrane. This method is described, for example, by Rhodes et al. (2007) Met
Hods in Cell Biology 82:309, the disclosure of which is incorporated herein by reference.

Microvesicles represent another carrier that can be used to introduce nucleic acids encoding the peptides or proteins described herein into host cells for recombinant expression.
In some embodiments, microvesicles in which the glycoprotein VSV-G and a genome-modifying protein, such as a nuclease, are co-overexpressed are used to efficiently deliver proteins to cells, and then Site-specific cleavage of endogenous polynucleotide sequences may be catalyzed to cause the genome of the cell to become covalently integrated with a polynucleotide of interest, such as a gene or regulatory sequence. The use of such vesicles, also called Gesicles, to genetically modify eukaryotic cells has been described, for example, by Quinn et al.
Genetic Modification of Target Cells by Direct Delivery of Active Protein [abst
ract]. In: Methylation changes in early embryonic genes in cancer [abstract], in
: Proceedings of the 18th Annual Meeting of the American Society of Gene and Cel
l Therapy; 2015 May 13, Abstract No. 122.

Viral Vectors for Nucleic Acid Delivery Viral genomes include the CXCR2 agonists described herein, including Gro-β, Gro-β T, Gro-β N69D, Gro-β T N65D, and variants thereof. There is a rich source of vectors that can be used to efficiently deliver foreign nucleic acids encoding peptides and proteins into host cells for recombinant expression. Viral genomes are particularly useful vectors for gene delivery. This is because polynucleotides contained within such a genome may be integrated into the genome of a cell, for example, by performing generalized or specialized transduction. These processes may occur as part of the viral vector's natural replication cycle and do not require the addition of proteins or reagents to induce genetic integration. Examples of viral vectors that may be used to introduce nucleic acid molecules encoding the peptides or proteins described herein into host cells for recombinant expression include parvoviruses, e.g. viruses (AAV), retroviruses, adenoviruses (e.g. Ad5, Ad26, Ad34, A
d35, and Ad48), coronaviruses, -stranded RNA viruses such as orthomyxoviruses (e.g. influenza viruses), rhabdoviruses (e.g. rabies and vesicular stomatitis viruses), paramyxoviruses (e.g. measles and Sendai), + Stranded RNA viruses such as picornaviruses and alphaviruses, as well as double-stranded DNA viruses such as adenoviruses, herpesviruses (e.g., herpes simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus), and pox Viruses (e.g. vaccinia, modified vaccinia Ankara (MVA), fowlpox virus and canarypox virus)
etc.), are included. Other viruses useful for delivering polynucleotides encoding peptides or proteins described herein to host cells for recombinant expression include, for example, Norwalk virus, Togavirus, Flavivirus. , reovirus,
Includes papovavirus, hepadnavirus, and hepatitis virus. Examples of retroviruses include avian leukemia-sarcoma, mammalian type C and B viruses, type D viruses, HTLV-BLV group,
Lentivirus, spumavirus (Coffin, JM, Retroviridae: The viruses and thei
r replication, In Fundamental Virology, Third Edition, BN Fields, et al., Eds
., Lippincott-Raven Publishers, Philadelphia, 1996). Other examples include murine leukemia virus, murine sarcoma virus, murine breast cancer virus, bovine leukemia virus, feline leukemia virus, feline sarcoma virus, avian leukemia virus, human T-cell leukemia virus, baboon endogenous virus, and gibbon ape leukemia virus. , Masson-Pfizer simian virus, simian immunodeficiency virus, simian sarcoma virus, Rous sarcoma virus and lentiviruses. Examples of other vectors are described, for example, in US Pat. No. 5,801,030, which is incorporated herein by reference for gene delivery and expression of recombinant proteins and peptides.

Treatment Methods As described herein, hematopoietic stem cell transplantation therapy involves the implantation of one or more blood cell types, such as a defective or defective blood cell lineage, in a patient suffering from a stem cell disorder. It may be applied to subjects in need of treatment, such as replantation or replantation. Hematopoietic stem and progenitor cells exhibit multipotency and therefore include, but are not limited to, granulocytes (e.g.
promyelocytes, neutrophils, eosinophils, basophils), red blood cells (e.g. reticulocytes, red blood cells), platelets (th
rombocytes) (e.g., megakaryoblasts, platelet-producing megakaryocytes, platelets), monocytes (e.g., monocytes, macrophages), dendritic cells, microglia, osteoclasts, and lymphocytes (e.g., can differentiate into several different blood lineages, including NK cells, B-cells, and T-cells). Hematopoietic stem cells are also capable of self-renewal and, therefore, can give rise to daughter cells that have the same potential as the mother cell and can be reintroduced into the transplant recipient, upon which they home in the hematopoietic stem cell niche. , reestablishing productive and sustained hematopoiesis. Hematopoietic stem and progenitor cells thus represent a useful therapeutic modality for treating a wide range of diseases in patients lacking or deficient in certain cell types of the hematopoietic lineage. This deficiency or defect may be caused by a reduction in the population of endogenous cells of the hematopoietic system, e.g., due to the administration of chemotherapeutic agents (e.g., in blood cancers such as those described herein). (in the case of patients suffering from cancer). Said defect or defect may be caused by a reduction in the population of endogenous hematopoietic cells, for example due to the activity of autoreactive immune cells such as T-lymphocytes or B-lymphocytes that cross-react with self-antigens. (eg, in the case of patients suffering from autoimmune disorders such as those described herein). Additionally or alternatively, defects or defects in cellular activity may be caused by aberrant expression of enzymes (e.g., in patients suffering from various metabolic disorders such as those described herein). (in cases of patients with

Therefore, in a patient who is defective or deficient in one or more cell types of the hematopoietic lineage, the defective or deficient cell population can be reconstituted in vivo, thereby eliminating the defect or deficiency in the endogenous blood cell population. Hematopoietic stem cells may be administered to treat conditions associated with depletion.
Hematopoietic stem cells and progenitor cells can be used, for example, to treat hemoglobin disorders selected from the group consisting of non-malignant hemoglobinopathies (e.g., sickle cell anemia, thalassemia, Fanconi anemia, aplastic anemia, and Wiskott-Aldrich syndrome). abnormalities) can be treated. In these cases, for example, CXCR4 antagonists and/or CXCR2 agonists may be administered to the donor (e.g., in response to such treatment, populations of hematopoietic stem and progenitor cells are likely to be released from stem cell niches such as bone marrow into circulating peripheral blood). It may be administered to a donor who has become a donor. The hematopoietic stem and progenitor cells thus mobilized are then harvested from the donor and administered to the patient. In the patient, the cells can home to the hematopoietic stem cell niche and reconstitute the population of cells that are damaged or missing in the patient.

Additionally or alternatively, hematopoietic stem and progenitor cells can be used to treat immune deficiencies such as congenital immunodeficiencies. Additionally or alternatively, the compositions and methods described herein can be used to treat an acquired immunodeficiency (e.g., an acquired immunodeficiency selected from the group consisting of HIV and AIDS). . In these cases, for example, CXCR4 antagonists and/or
Donors with CXCR2 agonists (e.g., donors whose populations of hematopoietic stem and progenitor cells are susceptible to release from stem cell niches such as the bone marrow into circulating peripheral blood in response to such treatment)
It may be administered to The hematopoietic stem and progenitor cells thus mobilized are then harvested from the donor and administered to the patient. In the patient, the cells home to the hematopoietic stem cell niche and are associated with immune cells (e.g., T-lymphocytes, B-lymphocytes, NK cells, or other immune cells) that are damaged or missing in the patient. ) can be reconstituted.

Using hematopoietic stem cells and progenitor cells to treat metabolic disorders (e.g., metabolic disorders selected from the group consisting of glycogen storage diseases, mucopolysaccharidoses, Gaucher disease, Hurler disease, sphingolipid and metachromatic leukodystrophy). It can also be treated. In these cases, for example, CXCR4
Administering an antagonist and/or a CXCR2 agonist to a donor (e.g., a donor that is susceptible to release of a population of hematopoietic stem and progenitor cells from a stem cell niche, such as the bone marrow, into circulating peripheral blood in response to such treatment) Sometimes. The hematopoietic stem and progenitor cells thus mobilized are then harvested from the donor and administered to the patient. In the patient, the cells can home to a hematopoietic stem cell niche and reconstitute a population of hematopoietic cells that are damaged or missing in the patient.

Additionally or alternatively, hematopoietic stem or progenitor cells can be used to treat malignancies or proliferative diseases (eg, hematological cancers or myeloproliferative diseases). In the case of cancer therapy, for example, CXCR4 antagonists and/or CXCR2 agonists may be released into the circulating peripheral blood from stem cell niches such as the bone marrow by populations of hematopoietic stem and progenitor cells in response to such therapy. It may be administered to donors who are more susceptible to The hematopoietic stem and progenitor cells thus mobilized are then harvested from the donor and administered to the patient. In the patient,
The cells home to a hematopoietic stem cell niche and are comprised of a population of hematopoietic cells that is damaged or missing in the patient (e.g., damaged by administering one or more chemotherapeutic agents to the patient). or a deficient hematopoietic cell population). In some embodiments, hematopoietic stem or progenitor cells may be infused into a patient to repopulate a population of cells that have been reduced during eradication of cancer cells, such as during systemic chemotherapy. . Exemplary blood cancers that can be treated by the methods of administering hematopoietic stem and progenitor cells according to the compositions and methods described herein include acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, leukemia, chronic lymphocytic leukemia, multiple myeloma, diffuse large cell type B-
and other cancer conditions, including cellular lymphoma and non-Hodgkin's lymphoma, as well as neuroblastoma.

The hematopoietic stem or progenitor cells mobilized into the peripheral blood of a subject can be obtained by any suitable technique.
It may be taken (e.g., recovered or collected) from the subject. For example, the hematopoietic stem cells or progenitor cells may be collected by blood sampling. In some embodiments, hematopoietic stem or progenitor cells mobilized into the peripheral blood of a subject as contemplated herein may be recovered (i.e., harvested) by using apheresis. . In some embodiments, apheresis may be used to enrich donor blood with mobilized hematopoietic stem or progenitor cells.

Additional diseases that may be treated by administering hematopoietic stem and progenitor cells to patients include, but are not limited to, adenosine deaminase deficiency and severe combined immunodeficiency, hyperimmunoglobulin M syndrome, Chediak- East disease, hereditary lymphohistiocytosis, osteopetrosis, osteogenesis imperfecta, storage disease, thalassemia major, systemic sclerosis, systemic lupus erythematosus
(systemic lupus erythematosus), multiple sclerosis, and juvenile rheumatoid arthritis.

Additionally, administration of hematopoietic stem and progenitor cells can be used to treat autoimmune diseases. In some embodiments, upon infusion into a patient, the transplanted hematopoietic stem and progenitor cells may home to a stem cell niche, such as bone marrow, and establish productive hematopoiesis. This, in turn, reduces the amount of autoimmune cell elimination that may occur through activation of autoreactive lymphocytes (e.g., autoreactive T-lymphocytes and/or autoreactive B-lymphocytes). Populations of cells can be reconstituted. Autoimmune diseases that can be treated by administering hematopoietic stem and progenitor cells to patients include, but are not limited to, psoriasis, psoriatic arthritis,
Type diabetes (type 1 diabetes), rheumatoid arthritis (RA), human systemic lupus erythematosus (SLE), multiple sclerosis (MS), inflammatory bowel disease (IBD), lymphocytic colitis, acute disseminated encephalomyelitis (ADEM), Addison's disease, generalized alopecia, ankylosing spondylitis, antiphospholipid antibody syndrome (APS), aplastic anemia, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease ( AIED), autoimmune lymphoproliferative syndrome (ALPS), autoimmune oophoritis, Barrow disease, Behcet's disease, bullous pemphigoid, cardiomyopathy, Chagas disease, chronic fatigue immunodeficiency syndrome (CFIDS), chronic inflammatory prolapse syndrome. Medullary polyneuropathy, Crohn's disease, cicatricial pemphigoid, celiac sprue dermatitis herpetiformis, cold agglutinin disease, CREST syndrome, Degos disease, discoid lupus erythematosus, autonomic neuropathy, endometriosis, mixed essential type cryoglobulinemia, fibromyalgia-fibromyositis, Goodpasture syndrome, Graves' disease, Guillain-Barre syndrome (GBS), Hashimoto's thyroiditis, hidradenitis suppurativa, idiopathic and/or acute thrombocytopenic purpura, Idiopathic pulmonary fibrosis, IgA neuropathy, interstitial cystitis, juvenile arthritis, Kawasaki disease, lichen planus, Lyme disease, Meniere's disease, mixed connective tissue disease (MCTD), myasthenia gravis, neuromyotonia, opso Clonus-myoclonus syndrome (OMS), optic neuritis, Ord's thyroiditis, pemphigus vulgaris, pernicious anemia, polychondritis,
polymyositis and dermatomyositis, primary biliary cirrhosis, polyarteritis nodosa, polyendocrine syndrome,
polymyalgia rheumatica, primary agammaglobulinemia,
Raynaud's phenomenon, Reiter's syndrome, rheumatic fever, sarcoidosis, scleroderma, Sjögren's syndrome, stiff person syndrome, Takayasu's arteritis, temporal arteritis (also known as giant cell arteritis), ulcerative colitis, collagen fibers These include genital colitis, uveitis, vasculitis, vitiligo, vulvodynia (vulvar vestibulitis), and Wegener's granulomatosis.

In some embodiments, methods of harvesting hematopoietic stem cells from a human subject are provided. The method includes administering a CXCR2 agonist and a CXCR4 antagonist to the human subject and recovering the hematopoietic stem cells from peripheral blood of the human subject.

In some embodiments, methods of transplanting hematopoietic stem cells into a human patient in need of a hematopoietic stem cell transplant are provided. The method includes administering a CXCR2 agonist and a CXCR4 antagonist to a hematopoietic stem cell donor, recovering the hematopoietic stem cells from peripheral blood of the donor, and transplanting the recovered hematopoietic stem cells into the patient. .

Donor and Patient Selection In some embodiments, said patient is said donor. In such cases, harvested hematopoietic stem or progenitor cells may be reinfused into the patient, so that the cells then home to hematopoietic tissue and establish productive hematopoiesis, thereby A set of cells that are defective or missing in the patient (e.g. megakaryocytes, thrombocytes)
, platelets, red blood cells, mast cells, myoblasts, basophils, neutrophils, eosinophils, microglia, granulocytes, monocytes, osteoclasts, antigen presenting cells, macrophages, dendritic cells, natural - Populations of killer cells, T-lymphocytes, and B-lymphocytes) may be engrafted or re-engrafted. In this scenario, the transplanted hematopoietic stem or progenitor cells are least likely to undergo graft rejection. This is because the injected cells are derived from the patient and express the same HLA class I and class II antigens that the patient expresses.

Alternatively, the patient and the donor may be different. In some embodiments, the patient and the donor may be related, eg, HLA matched. As described herein, HLA-matched donor-recipient pairs have a low risk of graft rejection. This is because endogenous T-cells and NK cells within the transplant recipient are less likely to recognize the incoming hematopoietic stem or progenitor cell graft as foreign and, as a result, mount an immune response against it. This is because the possibility is low. Exemplary HLA-matched donor-recipient pairs include genetically related donors and recipients, such as familial donor-recipient pairs (e.g., sibling donor-recipient pairs). It is.

In some embodiments, the patient and the donor are HLA-incompatible, which means that the donor and the recipient are HLA-incompatible with respect to at least one HLA antigen, particularly HLA-A, HLA-B, and HLA-DR. Occurs when there is a mismatch between To reduce the likelihood of graft rejection, for example, one haplotype may be matched between the donor and the recipient, and the other may be incompatible.

CD34 ^DIMM Cells When the donor and the patient are different, methods of treatment with hematopoietic ^stem cells, including CD34 ^DIMS , are based, in part, on the allo-reactive T
-It can inhibit the proliferation of lymphocytes, thereby preventing graft-versus-host disease (graft-versus-host disease).
rsus host disease (GVHD)).
'Aveni et al. (2015), supra.). In certain embodiments, administration of CXCR2 agonists and CXCR4 antagonists according to the methods disclosed herein mobilizes hematopoietic stem cells, including CD34 ^dim cells, from the bone marrow of the donor into peripheral blood. The CD34 ^dim cells are more abundant in peripheral blood compared to peripheral blood from non-mobilized mammals. In certain embodiments,
The CD34 ^dim cells are more present in peripheral blood than if the hematopoietic stem cells were mobilized using the CXCR4 antagonist alone.

Accordingly, the methods disclosed herein are useful for performing allogeneic hematopoietic stem cell transplantation in a patient in need thereof. For example, the method may include infusing into the patient a therapeutically effective amount of allogeneic hematopoietic stem cells, wherein the hematopoietic stem cells are isolated from human cells using the methods herein. It is mobilized from the donor's bone marrow into the peripheral blood of a human donor. In certain embodiments, the method includes: (i) about 50 μg/kg to about 1,000 μg/kg of a CXCR2 agonist selected from the group consisting of Gro-β, Gro-β T, and variants thereof;
and (ii) administering a CXCR4 antagonist to said donor.

Furthermore, in patients with standard-risk disease undergoing allogeneic hematopoietic cell transplantation, CD34 ^DIMM cells prolong overall survival (OS) and reduce non-relative mortality (OS).
apse mortality (NRM, i.e. time to death without relapse/relapse)) and infection (
For example, it has been shown to reduce the risk of cytomegalovirus (CMV) infection (Nakas
one et al. “CD34 ⁺ monocytes mobilized by G-CSF in donor PB and clinical outcomes
after all-HCT from related donors,” Poster presented at ^44th Annual Meeting of
the European Society for Blood and Marrow Transplantation, March 18-21, 2018, L
isbon, Portugal).

Accordingly, in certain embodiments, the methods of treating a stem cell disorder in a human patient disclosed herein include a therapeutically effective amount mobilized by any one of the methods disclosed herein. of hematopoietic stem cells into the patient, where the treatment increases OS, decreases NRM, and/or reduces the risk of infection (e.g., CMV infection). bring.

Additionally, the methods described herein may be used in preventing, reducing the risk of developing, or reducing the severity of post-transplant infections in patients in need thereof. The method may include infusing into the patient a therapeutically effective amount of hematopoietic stem cells, wherein the hematopoietic stem cells are from the human donor's bone marrow into the human donor's peripheral blood.
According to the methods described herein (e.g., (i) a CXCR2 agonist selected from the group consisting of Gro-β, Gro-β T, and variants thereof) at a concentration of about 50 μg/kg to about 1,000 μg/ (ii) administering a CXCR4 antagonist to the human donor) at a dose of 100 kg to the human donor. In certain embodiments, the infection is a CMV infection.

Furthermore, the present invention provides methods for preventing graft versus host disease (GVHD).
to reduce the risk of onset or reduce the severity of the disease in patients in need thereof;
Regarding the method. wherein the method includes infusing into the patient a therapeutically effective amount of hematopoietic stem cells, wherein the hematopoietic stem cells are transferred from the bone marrow of a mammalian donor into peripheral blood. (e.g., administering a CXCR2 agonist and a CXCR4 antagonist to said mammalian donor).

In certain embodiments, the hematopoietic stem and progenitor cells mobilized into the peripheral blood from the bone marrow of the donor are at least 1%, at least
2%, at least 5%, at least 10%, at least 15%, at least 20%, or at least 20%
or more CD34 ^dim cells. In certain embodiments, hematopoietic stem and progenitor cells mobilized from donor bone marrow into peripheral blood are about 1% to about 5%, about 1% compared to peripheral blood from non-mobilized mammals. % to approx. 10%, approx. 1% to approx. 15%, approx. 1% to approx. 20%, approx. 2% to approx. 25%
, 2% to about 5%, about 2% to about 10%, about 2% to about 15%, about 2% to about 20%, about 2% to about 25%, about 5% to about 10%, about 5
% to about 15%, about 5% to about 20%, about 5% to about 25%, about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about
15% to about 20%, contains about 15% to about 25% CD34 ^dim cells.

In certain embodiments, hematopoietic stem and progenitor cells mobilized from donor bone marrow into peripheral blood are at least 1.5 times, at least 2 times, at least 3 times more concentrated than peripheral blood from non-mobilized mammals. times, at least 4 times, at least 5 times, at least 6 times, at least 7 times
at least 8x, at least 9x, at least 10x, at least 11x, at least 12x, at least 13x, at least 14x, at least 15x, at least 20x, at least 30x, at least 50x more CD34 ^dim cells including. In certain embodiments, hematopoietic stem and progenitor cells mobilized from donor bone marrow into peripheral blood are about 1.5- to 30-fold, about 5-fold, compared to peripheral blood from an unmobilized mammal. ～about 25 times, about 10 times to about 20 times, or about 12 times to about 17 times
Contains twice as many CD34 ^dim cells.

In certain embodiments, the hematopoietic stem and progenitor cells mobilized from donor bone marrow into peripheral blood are at least 1%, at least 2% greater than when said hematopoietic stem cells were mobilized using said CXCR4 antagonist alone. , comprising at least 5%, at least 10%, at least 15%, at least 20%, or at least 20% or more CD34 ^dim cells. In certain embodiments, hematopoietic stem and progenitor cells mobilized into peripheral blood from the bone marrow of a donor combine said hematopoietic stem cells with said CX
about 1% to about 5%, about 1% to about 10%, about 1% more than when mobilized using CR4 antagonist alone
～about 15%, about 1% to about 20%, about 2% to about 25%, 2% to about 5%, about 2% to about 10%, about 2% to about 15%, about 2% to about 2
0%, about 2% to about 25%, about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 10% to about 15
%, about 10% to about 20%, about 10% to about 25%, about 15% to about 20%, about 15% to about 25% CD34 ^dim cells.

In certain embodiments, the hematopoietic stem and progenitor cells mobilized from donor bone marrow into peripheral blood are at least 1.5 times, at least 2 times as large as when said hematopoietic stem cells were mobilized using said CXCR4 antagonist alone. , at least 3 times, at least 4 times, at least 5 times,
At least 6x, at least 7x, at least 8x, at least 9x, at least 10x, at least 11x, at least 12x, at least 13x, at least 14x, at least 15x, at least 20x, at least 30x, at least 50x Contains twice as many CD34 ^dim cells. In certain embodiments, the hematopoietic stem and progenitor cells mobilized from donor bone marrow into peripheral blood are about 1.5-fold greater than when said hematopoietic stem cells were mobilized using said CXCR4 antagonist alone.
30 times, about 5 times to about 25 times, about 10 times to about 20 times, or about 12 times to about 17 times more CD34 ^dim cells.

Methods of genetically modifying hematopoietic stem and progenitor cells Hematopoietic stem cells obtained from a donor (or their progeny) prior to infusion into a patient, such as a patient with one or more stem cell disorders described herein may be genetically modified, for example, by disrupting endogenous genes. This strategy can be used, for example, to silence the expression of one or more major histocompatibility complex genes in hematopoietic stem cells that are allogeneic to the patient, thereby preventing graft rejection upon transplantation. It may reduce the possibility.

A wide range of methods have been established to disrupt target genes in cell populations.
In some embodiments, one such method involves the use of clustered regularly spaced cells, a system that originally evolved as an adaptive defense mechanism in bacteria and archaea against viral infections. short palindromic repetitions (clustered regularly interspac)
ed short palindromic repeats (CRISPR))/Cas system. Said
The CRISPR/Cas system includes palindromic repeat sequences within plasmid DNA and an associated Cas9 nuclease. This collection of DNA and proteins directs site-specific DNA cleavage at target sequences by first incorporating foreign DNA into the CRISPR locus. These foreign sequences and C
The polynucleotide containing the repeat-spacer elements of the RISPR locus is then transcribed in the host cell to generate a guide RNA, which subsequently anneals to the target sequence and binds the Cas
9 nuclease can be localized to this site. In this way, highly site-specific cas9-mediated DNA cleavage is caused in the foreign polynucleotide, as RNA:DNA hybridization dominates the interaction that brings cas9 into the vicinity of the target DNA molecule. It can be done. As a result, CRI to cut any target DNA molecule of interest
SPR/Cas systems can be designed theoretically. This technique was developed to edit the genome of eukaryotic cells (Hwang et al. (2013) Nature Biotechnology 31:227, the disclosure of which is incorporated herein by reference), and This technique may be used, for example, as an efficient method to site-specifically edit the genome of hematopoietic stem cells to cleave the DNA before incorporating the gene encoding the target protein. CRISPR/Cas
is described in US 8,697,359, for example,
This disclosure is incorporated herein by reference. Alternative methods for site-specific cleavage of genomic DNA prior to uptake of genes of interest in hematopoietic stem cells include zinc finger nucleases (ZFNs) and transcriptional activation. This includes the use of transcription activator-like effector nucleases (TALENs). Unlike the CRISPR/Cas system, these enzymes do not contain guide polynucleotides to localize to specific target sequences. Target specificity is instead controlled by DNA binding domains within these enzymes. For the use of ZFNs and TALENs in genome editing applications, see, for example, Urnov et al. (2010) Nature R
eviews Genetics 11:636; and Joung et al. (2013) Nature Reviews Molecular Cell B
iology 14:49, the disclosures of both of which are incorporated herein by reference.

Additional genome editing techniques that can be used to integrate polynucleotides encoding target genes into the genome of hematopoietic stem cells include those that can be rationally designed to site-specifically cleave genomic DNA. Includes using ARCUS ^TM meganuclease. The use of these enzymes to integrate genes encoding target genes into the genome of mammalian cells relies on the fact that certain structure-activity relationships have been established for such enzymes. It is advantageous in To create a nuclease that selectively cuts DNA at a desired location and enables site-specific integration of a target gene into the nuclear DNA of hematopoietic stem cells, a single-stranded meganuclease is May be modified at amino acid positions. These single-stranded nucleases are described, for example, in US 8,021,867 and US 8,4
45,251, the disclosures of each of which are incorporated herein by reference.

Kinetics for Dosing CXCR2 Agonists and CXCR4 Antagonists In cases where the donor is administered both a CXCR4 antagonist and a CXCR2 agonist, the two drugs may be administered to the donor at the same time. In some embodiments, the CXCR4 antagonist and the CXCR2 agonist may be formulated together with each other and administered as the same pharmaceutical composition. Alternatively, the CXCR4 antagonist and the CXCR2 agonist may be formulated as separate pharmaceutical compositions and administered to the donor separately but simultaneously.

In some embodiments, the CXCR4 antagonist is administered to the donor prior to administering the CXCR2 agonist. In some embodiments, the CXCR4 antagonist is administered for about 30 minutes to about 180 minutes, such as about 40 minutes to about 160 minutes, about 5 minutes, before administering the CXCR2 agonist.
0 minutes to approximately 150 minutes, approximately 60 minutes to approximately 140 minutes, approximately 70 minutes to approximately 130 minutes, approximately 60 minutes to approximately 120 minutes, approximately 70 minutes to approximately 110 minutes, or approximately 80 minutes to approximately 100 minutes (for example, about 30 minutes, about 35 minutes before administering the CXCR2 agonist;
Approximately 40 minutes, approximately 45 minutes, approximately 50 minutes, approximately 55 minutes, approximately 60 minutes, approximately 65 minutes, approximately 70 minutes, approximately 75 minutes, approximately 80 minutes, approximately 85 minutes,
Approximately 90 minutes, approximately 95 minutes, approximately 100 minutes, approximately 105 minutes, approximately 110 minutes, approximately 115 minutes, approximately 120 minutes, approximately 125 minutes, approximately 130 minutes,
about 135 minutes, about 140 minutes, about 145 minutes, about 150 minutes, about 155 minutes, about 160 minutes, about 165 minutes, about 170 minutes, about 175 minutes, or about 180 minutes). In some embodiments, the CXCR4 antagonist is administered from about 30 minutes to about 60 minutes before administering the CXCR2 agonist (e.g., about 30 minutes, about 35 minutes, about 40 minutes before administering the CXCR2 agonist). (about 45 minutes, about 50 minutes, about 55 minutes, or about 60 minutes) to said donor. In some embodiments, the CXCR4 antagonist is
The CXCR2 agonist is administered to the donor approximately 45 minutes prior to administration.

Isolation of the population of hematopoietic stem cells or progenitor cells is performed for about 10 minutes to about 60 minutes (e.g., about 10 minutes to about 1.9 hours, about 20 minutes to about 1.8 hours) after the end of administration of the CXCR4 antagonist and the CXCR2 agonist. , about 25 minutes to about 1.7 hours, about 30 minutes to about 1.6 hours, about 40 minutes to about 1.5 hours (e.g., about 10 minutes, about 15 minutes, about 20 minutes after the end of administration of the CXCR4 antagonist and the CXCR2 agonist) minutes, about 25
minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes, about 60 minutes, or about 120 minutes)). In some embodiments, isolation of a population of hematopoietic stem or progenitor cells comprises
About 10 minutes to about 20 minutes after the end of the administration of the CXCR4 antagonist and the CXCR2 agonist (e.g., about 10 minutes, about 11 minutes after the end of the administration of the CXCR4 antagonist and the CXCR2 agonist)
minutes, approximately 12 minutes, approximately 13 minutes, approximately 14 minutes, approximately 15 minutes, approximately 16 minutes, approximately 17 minutes, approximately 18 minutes, approximately 19 minutes, or approximately 20 minutes)
may start. In some embodiments, isolation of a population of hematopoietic stem or progenitor cells begins about 15 minutes after the end of administration of said CXCR4 antagonist and said CXCR2 agonist.

In some embodiments, the population of hematopoietic stem or progenitor cells is grown for a period of about 15 minutes to about 6 hours, such as about 20 minutes to about 4.5 hours, about 30 minutes to about 4 hours, about 40 minutes to about 3.5 hours, about 50
minutes to about 3 hours, or about 1 hour to about 2 hours (for example, about 15 minutes, about 20 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes, about 60 minutes, about 65 minutes, about 70 minutes, about 75 minutes, about 80 minutes, about 85 minutes, about 90 minutes, about 95 minutes, about 100 minutes, about 105 minutes, about 110 minutes, about 115 minutes, about 120 minutes , about 180 minutes, about 240 minutes, about 300 minutes, or about 360 minutes) from the donor. In some embodiments,
Populations of hematopoietic stem and progenitor cells are incubated for a period of about 30 minutes to about 1 hour (e.g., about 30 minutes,
from the donor over a period of about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes, or about 60 minutes).

In some embodiments, the hematopoietic stem or progenitor cells may be collected by apheresis. In some embodiments, the hematopoietic stem or progenitor cells may be collected by collecting peripheral blood from the donor (ie, subject).

Routes of Administration for CXCR2 Agonists and CXCR4 Antagonists The CXCR4 antagonists and CXCR2 agonists described herein may be administered to a subject by a variety of routes (e.g., intravenous, subcutaneous, intramuscular, or parenteral). . The most suitable route of administration in any given case will depend on the particular drug being administered, the patient, the method of pharmaceutical formulation, the method of administration (e.g., time of administration and route of administration), age, weight, sex of the patient, and the disease being treated. depending on the severity of the disease, the patient's diet, and the patient's excretion rate. Preferably, the CXCR2 agonist (eg, Gro-β, Gro-β T, or a variant thereof) may be administered intravenously to the donor. Under these conditions, CXCR2 agonists as described herein inhibit CD34 ⁺ CD
It rapidly generates a population of cells enriched in 90 ⁺ CD45RA ⁻ cells (hematopoietic stem cells) and reduces the recruitment of other cell types (eg, leukocytes, neutrophils, lymphocytes, and monocytes). This property is described in further detail in Example 1 below.

Pharmaceutical Compositions The CXCR2 agonists and CXCR4 antagonists contemplated herein are each formulated into the same pharmaceutical composition for administration to a subject, such as a mammalian subject (e.g., a human subject). Sometimes. For example, contemplated herein are pharmaceutical compositions comprising a CXCR2 agonist and/or a CXCR4 antagonist in admixture with one or more suitable diluents, carriers, and/or excipients. Pharmaceutical compositions may include sterile aqueous suspensions. Conventional procedures and ingredients for selecting and preparing suitable formulations are described, for example, in Remington: The Science and Practical
e of Pharmacy(2012, ^22nd ed.) and The United States Pharmacopeia: The National Fo
rmulary (2015, USP 38 NF 33), the disclosure of which is incorporated herein by reference.

A pharmaceutical composition, alone or in combination with a pharmaceutically acceptable carrier, may be administered to a subject (e.g., a human subject) and the proportions of the active pharmaceutical ingredients (i.e., CXCR2 agonist and/or CXCR4 antagonist), the chosen route of administration, and standard pharmaceutical practice.

Administration and Dosing of CXCR2 Agonists and/or CXCR4 Antagonists The contemplated CXCR2 agonists and CXCR4 antagonists may be administered to a subject, such as a mammalian subject (e.g., a human subject), by one or more routes of administration. . For example, intended CXCR2 agonists and CXCR4 antagonists can be administered intravenously, intraperitoneally, intramuscularly, among others.
It may be administered to a subject by intraarterial or subcutaneous injection.

The intended CXCR2 agonist and CXCR4 antagonist may be administered to the subject at one or more doses. For example, CXCR2 agonists and/or CXCR4 antagonists may be administered as a single dose or in 2, 3, 4, 5, or more doses. If administered in multiple doses, subsequent doses may be administered on the same day as the first dose or within 1 day, 1 week, 1 month, or 1 year or more after administration. . For example, the contemplated CXCR2 agonists and CXCR4 antagonists described herein may be administered to a subject, such as a human subject, once a day, once a week, once a month, or once a year or more, e.g. Administration may depend on factors such as the subject's age, weight, sex, the subject's diet, and the subject's excretion rate. In certain embodiments, the contemplated CXCR2 agonist and CXCR4 antagonist are each
Administer in a single dose once daily.

The hematopoietic stem or progenitor cells and pharmaceutical compositions described herein may be administered to a subject in one or more doses. If multiple doses are administered, subsequent doses may be administered 1 day, 1 week, 1 month, or 1 year or more after the first dose is administered. For example, the hematopoietic stem cells and pharmaceutical compositions described herein can be used to treat one or more diseases described herein.
A subject, such as a human subject, suffering from a condition or disorder, for example, the subject's age, weight, gender,
It may be administered once a day, once a week, once a month, or more than once a year, depending on factors such as the severity of the disease being treated, the subject's diet, and the subject's excretion rate.

The following examples are presented to provide those skilled in the art with a description of how the compositions and methods described herein can be used, made, and evaluated, and are based solely on the present invention. It is intended to be illustrative of the invention and is not intended to limit the scope of what the inventors consider to be their invention.

Example 1. Effects of Gro-β T on hematopoietic stem cell mobilization in mice and rhesus monkeys Mobilized peripheral blood grafts are currently a major source of hematopoietic stem and progenitor cells (HSPCs) for both autologous and allogeneic transplantation. source of supply. The most common clinical hematopoietic stem cell mobilization protocol is filgrastim (G-CSF) for 5 days. This regimen requires daily injections, is associated with bone pain, and often has unexpectedly low yields. A rapid mobilization method that ideally requires only a single procedure and has robust and predictable kinetics would be a significant improvement over current standards of care. In mice, Gro-β T, a unique CXCR2 agonist, rapidly mobilizes stem and progenitor cells 15 minutes after a single injection. CX
When co-administered with the CR4 inhibitor plerixafor (AMD3100), recruitment is synergistically increased and grafts become enriched in long-term hematopoietic stem cells with increased engraftment potential (e.g.
FIG. 1A; see LT-HSC = Linc-kit ⁺ Sca-1 ⁺ CD150 ⁺ CD48 ⁺ ). Grafts containing cells recruited by Gro-β T and plerixafor had a higher relative number of competitive repopulation units at week 16 than grafts containing cells recruited by G-CSF alone.
(competitive repopulating unit) (CRU) (Figure 1B).

In this example, treatment with Gro-β T and AMD3100 in combination increases the recruitment of CD34 ⁺ cells and colony forming units ( We present data to demonstrate that CFU) is significantly enhanced.

As described below, hematopoietic stem cell mobilization was studied in rhesus macaques using Gro-β T and plerixafor (also called AMD3100).

Methods Male rhesus macaques were treated with AMD3100 alone or in combination with Gro-β T. Blood was collected immediately before treatment and 0.5, 1, 2, 4, and 24 hours after treatment and analyzed by multicolor flow cytometry to quantify HSPC numbers. Additional aliquots of mobilized blood were plated on methylcellulose plates and CFU were counted after 7 days.

Results Figure 2A shows the pharmacokinetic profile of various doses of Gro-β T when administered intravenously to rhesus monkeys. FIG. 2B shows the pharmacokinetic profile of various doses of Gro-β T when administered subcutaneously to rhesus monkeys. In both experiments, Gro-β T was administered to subjects at the same time as plerixafor.

As shown in Figures 3-6, Gro-β T and AMD3100 mobilize white blood cells into peripheral blood.
Animals were mobilized with AMD3100 alone or in combination with Gro-β T. peripheral blood. Peripheral blood was collected at the indicated time points and white blood cells were counted on a HESKA Hematology Analyzer. The total number of white blood cells, neutrophils, lymphocytes, and monocytes per 1 μL of peripheral blood was determined. Data shown in Figures 2-5 are expressed as mean ± SEM and represent 5 animals per group. Statistical significance was determined based on two-way analysis of variance and Dunnett's multiple comparison post hoc test (** p<0.01).

Importantly, as shown in Figures 7 and 8, Gro-β T and AMD3100 significantly reduce C into peripheral blood.
Induces strong recruitment of D34 ⁺ cells. The data shown in Figures 7 and 8 include the percentage of whole blood occupied by CD34 ⁺ hematopoietic stem and progenitor cells for each treatment group. Peripheral blood 1
Absolute numbers and fold changes of CD34 ⁺ cells per μL were quantified using a single platform quantification method. Data shown in Figures 7 and 8 are expressed as mean ± SEM and represent 5 animals per group. Statistical significance was determined based on two-way analysis of variance and Dunnett's multiple comparison post hoc test (* p<0.05, ** p<0.01, **** p<0.0001).

Furthermore, as shown in Figures 9 and 10, CD34 recruited in response to Gro-β T + AMD3100
⁺ cells are enriched with immature CD34 ⁺ CD90 + CD45RA ^- stem cells and progenitor cells. Figures 9 and 1
Data shown at 0 include the percentage of whole blood occupied by CD34 ⁺ CD90 ⁺ CD45RA ⁻ hematopoietic stem and progenitor cells for each treatment group. Absolute numbers and fold changes of CD34 ⁺ CD90 ⁺ CD45RA ⁻ cells per μL of peripheral blood are shown. Data shown in Figures 9 and 10 are expressed as mean ± SEM and represent 5 animals per group. Statistical significance was determined based on two-way analysis of variance and Dunnett's multiple comparisons post hoc test (* p<0.05.** p<0.01, *** p<0.001, **** p< 0.000
1).

Furthermore, as shown in FIG. 11, Gro-β T and AMD3100 mobilize hematopoietic stem cells and progenitor cells with colony-forming ability. The number of CFU per mL of peripheral blood was counted after 7 days of culture in methylcellulose. Data shown in Figure 11 are expressed as mean ± SEM and represent 3-5 animals per group. Statistical significance was determined based on two-way analysis of variance and Dunnett's multiple comparison post hoc test (*p<0.05). The ratio of MMP-9 to TIMP-1 is
and further increases after treatment with AMD3100 (Figures 12-14).

CD34 ⁺ cells in rhesus macaques using various doses of Gro-beta T and AMD3100 (e.g. CD34
Additional data summarizing the recruitment of ^{CD90 +} CD45RA ⁻ cells) are provided in Tables 8-11 below.
Quantities are listed in Tables 8-11 using the following notation: "median value (minimum value observed - maximum value observed)".

conclusion
A single treatment of Gro-β T in combination with AMD3100 induces robust mobilization of stem and progenitor cells within 4 hours of administration in non-human primates. Furthermore, compared to AMD3100 alone, Gro-β T in combination with AMD3100 resulted in 2-3 times more CD34 ⁺ CD90 ⁺ CD45RA ^- stem and progenitor cells, suggesting significantly improved graft quality. do.

Furthermore, these data demonstrate that when G-CSF is contraindicated or associated with adverse events, Gro-β T, when used in combination with AMD3100, is effective against sickle cell disease (SCD) and multiple sclerosis ( It has the potential to be a more reliable and safer alternative to G-CSF in autologous and allogeneic transplants, including in diseases such as MS).

Example 2. Hematopoietic stem cells mobilized with CXCR2 agonists and/or CXCR4 antagonists
Determining whether a population is suitable for ex vivo expansion and/or therapeutic use Using the compositions and methods described herein, one skilled in the art can A population of hematopoietic stem or progenitor cells may be mobilized in a donor. In some embodiments, the practitioner releases a population of hematopoietic stem cells into circulating peripheral blood while decreasing the recruitment of other cells of the hematopoietic system, such as leukocytes, neutrophils, lymphocytes, and monocytes. The CXCR4 antagonist and CXCR2 agonist can be administered in amounts sufficient to cause .

When administering a CXCR4 antagonist in combination with a CXCR2 agonist, the physician may administer the two agents to the donor at the same time or at different times. In some embodiments, the CXCR4 antagonist is administered from about 30 minutes to about 180 minutes before administration of the CXCR2 agonist.
For example, about 40 minutes to about 160 minutes, about 50 minutes to about 150 minutes, about 60 minutes to about 140 minutes, about 70 minutes to about 130 minutes, about 60 minutes
minutes to about 120 minutes, about 70 minutes to about 110 minutes, or about 80 minutes to about 100 minutes (e.g., about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes before administration of the CXCR2 agonist) minutes, about 55 minutes, about 60 minutes, about 65 minutes, about 70 minutes,
Approximately 75 minutes, approximately 80 minutes, approximately 85 minutes, approximately 90 minutes, approximately 95 minutes, approximately 100 minutes, approximately 105 minutes, approximately 110 minutes, approximately 115 minutes, approximately 12
0 minutes, approximately 125 minutes, approximately 130 minutes, approximately 135 minutes, approximately 140 minutes, approximately 145 minutes, approximately 150 minutes, approximately 155 minutes, approximately 160 minutes, approximately 1
65 minutes, about 170 minutes, about 175 minutes, or about 180 minutes).

To evaluate the effectiveness of the mobilization regimen, peripheral blood samples may be isolated from the subject after administration of the CXCR2 agonist and/or CXCR4 antagonist. The sample is then subjected to one or more parameters of the sample (e.g., the parameters listed in Table 2).
) may be characterized by obtaining input values for each of them. Exemplary parameters that may be used to assess the effectiveness of a regimen to mobilize hematopoietic stem cells include the ratio of hematopoietic stem cells to other cell types (e.g., leukocytes, neutrophils, lymphocytes, and monocytes);
and the relative frequency of hematopoietic stem cells in the sample. Input values for these parameters can be used, for example, by flow cytometry and fluorescence activated cell sorting.
immunological phenotyping methods known in the art, such as FACS (FACS) techniques.

When obtaining and analyzing input values for two or more parameters listed in Table 2, combinations of parameters may be analyzed. In some embodiments, in a sample obtained from a donor's peripheral blood after administration of a CXCR2 agonist and/or a CXCR4 antagonist,
The ratio of hematopoietic stem cells to white blood cells, the ratio of hematopoietic stem cells to neutrophils, the ratio of hematopoietic stem cells to lymphocytes, the ratio of hematopoietic stem cells to monocytes, and/or the relative frequency of hematopoietic stem cells may be analyzed. For example, combinations of parameters listed in any one of Tables 3-6 may be analyzed.

Once input values are obtained for each of the one or more parameters, the input value(s) may then be compared to a reference standard for each parameter. If the above reference criteria are fulfilled (
For example, if the ratio of hematopoietic stem cells to other hematopoietic cell types is sufficiently high, or if the relative frequency of hematopoietic stem cells in a sample obtained from peripheral blood of said donor is sufficiently high, said cells may be removed ex vivo. and/or for therapeutic use.

Example 3. Treatment of hematological disorders by administering hematopoietic stem cells or progenitor cell grafts
Using the compositions and methods described herein, one skilled in the art can treat the hematological conditions described herein by administering hematopoietic stem or progenitor cell grafts to a patient. Stem cell disorders such as: For example, a practitioner identifies a human donor of hematopoietic stem or progenitor cells that is likely to respond to a mobilization regimen, as outlined in Example 1, and then, for example, as described in Example 2. Populations of hematopoietic stem or progenitor cells may be mobilized. After mobilization, the physician may isolate a population of hematopoietic stem or progenitor cells from the donor. The isolation of said cells may be carried out using, for example, a CXCR4 antagonist and/or a CXCR2
about 10 minutes to about 60 minutes after the end of administration of the agonist, such as about 15 minutes to about 55 minutes, about 20 minutes to about 50 minutes, about 25 minutes to about 45 minutes, or about 30 minutes to about 40 minutes (for example, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes after the end of administration of the CXCR4 antagonist and CXCR2 agonist) , about 50 minutes, about 55 minutes, or about 60 minutes).

The isolation procedure can be carried out for about 15 minutes to about 6 hours, such as about 20 minutes to about 4.5 hours, about 30 minutes to about 4 hours, about 40 minutes.
over a period of minutes to about 3.5 hours, about 50 minutes to about 3 hours, or about 1 hour to about 2 hours (e.g., about 1
5 minutes, about 20 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes, about 60 minutes, about 65 minutes, about 7
0 minutes, about 75 minutes, about 80 minutes, about 85 minutes, about 90 minutes, about 95 minutes, about 100 minutes, about 105 minutes, about 110 minutes, about 115 minutes, or about 120 minutes) be. In some embodiments, the population of hematopoietic stem and progenitor cells is grown over a period of about 30 minutes to about 1 hour (e.g., about 30 minutes, about 35 minutes, about
from the donor over a period of 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes, or about 60 minutes).

Following this isolation process, the patient may then receive an infusion (eg, intravenous infusion) of mobilized, isolated hematopoietic stem or progenitor cells. The patient may be the donor, or a patient who is HLA matched to the donor, thereby reducing the likelihood of graft rejection. The patient may be, for example, a patient suffering from a cancer such as a blood cancer described herein. Additionally or alternatively, the patient may be suffering from an autoimmune disease or metabolic disorder as described herein. ^Physicians may collect mobilized and isolated hematopoietic stem ^{or progenitor} cells ^, e.g. 10 ⁷ CD34 ⁺ cells/kg, approximately 2 x ^{10 5} CD34 ⁺ cells
/kg ~ approx. 9 x 10 ⁶ CD34 ⁺ cells/kg, approx. 3 x 10 ⁵ CD34 ⁺ cells/kg ~ approx. 8 x 10 ⁶ CD34 ⁺ cells/kg, approx. 4 x 10
⁵ CD34 ⁺ cells/kg to about 7 x 10 ⁶ CD34 ⁺ cells/kg) depending on various factors such as patient age, weight, and severity of the disease being treated. There is.

The physician may, for example, collect blood samples from the patient after administering the implant and collect hematopoietic stem cells or cells of the hematopoietic lineage (e.g., megakaryocytes, thrombocytes).
tes), platelets, red blood cells, mast cells, myoblasts, basophils, neutrophils, eosinophils, microglia, granulocytes, monocytes, osteoclasts, antigen presenting cells, macrophages, dendritic cells ,
Engraftment of the hematopoietic stem cell transplant may be monitored by determining an increase in natural killer cells, T-lymphocytes, B-lymphocytes, etc.). This analysis
For example, about 1 hour to about 6 months, about 2 hours to about 5 months, about 3 hours to about 4 months, about 4 hours to about 3 months, about 10 hours to about 7 days, about 24 hours after hematopoietic stem cell transplantation therapy. hours to about 96 hours, or more (e.g.
1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 1
2 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 2
2 hours, 23 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks , 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16
It may take place every week, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, or more. The concentration of hematopoietic stem cells or cells of the hematopoietic lineage is increased after transplantation therapy compared to the concentration of the corresponding cell type before transplantation therapy (e.g., 1%, 2%, 3%, 4%, 5%, 6 %, 7
%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 500%, or more) provides one indication that the hematopoietic stem or progenitor cell transplantation therapy is effective in treating stem cell disorders.

Example 4: Activation of CD34 ^dim cells using CXCR2 agonists and/or CXCR4 antagonists
In this example, we present data showing that combination treatment with Gro-β T and AMD3100 (plerixafor) significantly reduced CD34 ^dim compared to that achieved with AMD3100 alone in non-human primates (NHPs).
We demonstrate that cell recruitment is significantly enhanced.

Cells were mobilized according to the method of Example 1. Peripheral blood samples were collected for CD34, CD11b, CD14, C
D16, CD45, and SSC markers were tested using a flow cytometer.

As shown in Figure 15, intravenous administration of 450 μg/kg Gro-β T and subcutaneous administration of 1 mg/kg AMD3100 surprisingly results in the recruitment of a population of cells that are CD34 ^dim CD11b ⁺ . Subpopulations within this population are CD14 ⁺ CD16 ⁺ , CD14 ^- CD16 ^- , and combinations thereof. Figure 16 shows CD3
4 shows that the recruitment of ^dim cells is significantly enhanced when Gro-β T and AMD3100 are administered in combination compared to AMD3100 alone.

Example 5: Gro-β T in combination with AMD3100 is effective against graft versus host disease.
This example demonstrates how co-treatment with Gro-β T and AMD3100 (plerixafor) resulted in the recruitment of CD34 ^dim cells, which have immunosuppressive properties. By becoming significantly enhanced and transplanting this cell population, we reduced the incidence of acute graft versus host disease (aGVHD) in a xenograft GVHD mouse model. We demonstrate a significant reduction in CD34 ^dim cells compared to transplantation of peripheral blood mononuclear cells with lower concentrations of CD34 dim cells.

Rhesus macaques were treated with Gro-β T, AMD3100, Gro-β T and AMD3100, or G-CSF alone. Peripheral blood samples were tested using flow cytometry. Gro-β T and AM
D3100-mobilized animals showed a >15-fold increase in CD34 ^dim cell numbers over baseline at 4 hours post-treatment (p<0.0001, n=13, Figure 17). In contrast, animals subjected to mobilization with G-CSF alone had only an approximately 9-fold increase in CD34 ^dim cell numbers over baseline 4 hours after the final dose of G-CSF (p<0.0001, n= 3, Figure 17).

Furthermore, the composition of unmobilized cells and grafts mobilized with G-CSF, Gro-β T and AMD3100, and AMD3100 alone is shown in FIG. As shown, grafts mobilized using Gro-β T and AMD3100 showed a 3-fold increase in CD34 ^dim cells and a 3-fold increase in T-cells compared to grafts mobilized using G-CSF. Showing a 3-fold increase.

To determine whether CD34 ^dim cells have immunosuppressive properties, we tested them with Gro-β
Sorted from the peripheral blood of rhesus macaques treated with T and AMD3100. CD34 ^dim cells, in
Cocultured with carboxyfluorescein succinimidyl ester (CFSE)-labeled autologous T-cells stimulated with anti-CD2/CD3/CD28-coated beads in vitro. As shown in Figure 19, Gro-β
CD34 ^dim cells mobilized with T and AMD3100 suppressed T-cell proliferation as measured by CFSE staining after 4 days.

To assess whether these immunosuppressive cells prevent GVHD, we developed a xenograft GVHD model in immunodeficient NOD scid gamma (NSG) mice. Gro-β
Peripheral blood (6 x ¹⁰⁶ PBMCs) mobilized with T and AMD3100 and containing a high proportion of CD34 ^dim cells was injected into sublethally irradiated NSG mice. This was compared to unmobilized primate PBMCs (6 x 10 ⁶ PBMCs) and G-CSF mobilized primate PBMCs (6 x ^{10 6} PBMCs), which contained relatively low numbers of CD34 ^dim cells. As shown in the survival curves in Figure 20, on day 24, all mice (13/13) transplanted with non-mobilized PBMCs died of aGVHD, whereas for comparison, mice transplanted with PBMCs mobilized with AMD3100 5/16 of the mice and 3/16 of the mice transplanted with G-CSF-mobilized PBMC died of aGVHD, as well as Gr.
No mice transplanted with o-β T and AMD3100 mobilized PBMCs died. At day 60 post-transplant, 15/16 of the mice transplanted with Gro-β T and AMD3100 remained alive, whereas only 10/16 of the mice transplanted with peripheral blood mobilized with AMD3100 remained alive. Only 11/16 mice transplanted with CSF-mobilized peripheral blood remained alive (see Figure 20). As shown in Figure 21A, mice transplanted with unmobilized PBMCs (“non-mobilized”) were compared to mice transplanted with PBMCs mobilized with Gro-β T and AMD3100 (“Gro-β T + plerixafor”). and showed more donor T-cells ("approximately 160 times", see Figure 21A) at day 14 post-transplant (*p<0.01, n=6-8). Furthermore, as shown in FIG. 21A, similarly, mice transplanted with PBMCs mobilized with AMD3100 (i.e., plerixafor) alone ("plerixafor") or mice transplanted with PBMCs mobilized with G-CSF alone ("plerixafor"). G-CSF') compared to mice transplanted with Gro-β T- and AMD3100-mobilized PBMCs, T-cells (approximately 60-fold higher ('p<0.05') and approximately 10-fold higher, respectively) High (“p<0.01”
))showed that.

To confirm that CD34 ^dim cells in grafts mobilized with Gro-β T and AMD3100 are involved in preventing aGvHD in the NSG mouse xenograft model shown in Figure 20, an additional cohort of mice was 6 x 10 ⁶ PBMCs from rhesus macaques mobilized with T and AMD3100
was transplanted. Here, first, CD34 ^dim cells were sorted using fluorescence activated cell sorting.
T-cell numbers and survival were compared to cohorts transplanted with Gro-β T and AMD3100-mobilized PBMCs and to cohorts transplanted with unmobilized PBMCs. Figure 2
As shown in 1B, 14 days after transplantation, mice transplanted with PBMCs mobilized with Gro-β T and AMD3100 (“Gro-β T + plerixafor”) were compared to mice transplanted with unmobilized PBMCs (“Gro-β T + plerixafor”). Non-mobilization”
), the number of rhesus CD45+ CD3+ T-cells in peripheral blood was significantly decreased (p<0.001,
n=10/group). In contrast, mice transplanted with Gro-β T- and AMD3100-mobilized CD34 ^dim cell-depleted PBMCs (n=10) compared to mice transplanted with unmobilized PBMCs (“non-mobilized”). ,
By day 14 post-transplant, there were no significant differences in T-cell numbers (see Figure 21B). As shown in Figure 21C, by day 44 after transplantation, mice transplanted with PBMCs mobilized with Gro-β T and AMD3100 (
``Gro-β T + plerixafor'') survived, whereas 0/10 of mice transplanted with unmobilized PBMCs (``non-mobilized'') survived (p<0.05); In addition, mice transplanted with PBMCs from which CD34 ^dim cells mobilized with Gro-β T and AMD3100 were removed (“Gro-β T + plerixafor CD34 ^{dim cells”)}
-removal"), only 4/10 survived (no significant difference compared to non-recruitment).

Therefore, these data demonstrate that co-administration of Gro-β T and AMD3100 not only rapidly and effectively mobilizes highly enriched HSCs, but also that these HSCs are more susceptible to mobilization than AMD3100 alone or current standards of care. is enriched with a population of CD34 ^dim cells that have potent immunosuppressive activity compared to G-CSF, and transplantation of this population of CD34 ^dim cells reduces the incidence or risk of aGVHD. This is proven.

Other Embodiments All publications, patents, and patent applications mentioned herein are as if each independent publication or patent application were specifically and individually indicated to be incorporated by reference. Incorporated herein by reference to the same extent.

Although the invention has been described in connection with specific embodiments thereof, the invention is susceptible to further modifications and this application generally describes the invention in accordance with the principles of the invention and which is well known in the art to which the invention pertains. Any modification, use, or It will be understood that adaptation is intended to be included.

Other embodiments are within the scope of the claims.

Claims (167)

A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
(i) a CXCR2 agonist selected from the group consisting of Gro-β, Gro-β T, and variants thereof at a dose of about 50 μg/kg to about 300 mg/kg; and (ii) a CXCR4 antagonist to said donor. A method comprising administering to 2. The method of claim 1, wherein the CXCR2 agonist is Gro-βT. 3. The method of claim 1 or 2, wherein the CXCR2 agonist is administered at a concentration of about 100 μg/kg to about 250 μg/kg.
A dose of μg/kg is administered to said donor. 4. The method of claim 3, wherein the CXCR2 agonist is administered at a concentration of about 125 μg/kg to about 225 μg/kg.
g doses to said donor. 5. The method of claim 4, wherein the CXCR2 agonist is administered to the donor at a dose of about 150 μg/kg. 6. The method of any one of claims 1-5, wherein said CXCR2 agonist is administered intravenously to said donor. 7. The method of any one of claims 1-6, wherein said CXCR4 antagonist is administered subcutaneously to said donor. The method of any one of claims 1 to 7, wherein the CXCR4 antagonist is plerixafor or a pharmaceutically acceptable salt thereof. 9. The method of claim 8, wherein said plerixafor or a pharmaceutically acceptable salt thereof is administered to said donor at a dose of about 50 μg/kg to about 500 μg/kg. 10. The method of claim 9, wherein said plerixafor, or a pharmaceutically acceptable salt thereof, is administered to said donor at a dose of about 200 μg/kg to about 300 μg/kg. 11. The method of claim 10, wherein said plerixafor or a pharmaceutically acceptable salt thereof is administered to said donor at a dose of about 240 μg/kg. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
a CXCR2 agonist and a CXCR4 antagonist sufficient to produce a population of cells in which, after administration of said CXCR2 agonist and said CXCR4 antagonist, the ratio of CD34+ cells to white blood cells in a sample of said donor's peripheral blood is from about 0.0008 to about 0.0021. administering to said donor an amount of said donor. 13. The method of claim 12, wherein the ratio of CD34+ cells to white blood cells in the sample is about 0.0010 to about 0.0018. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
CXCR2 agonists and CXCR4 antagonists were evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and CXCR4 method. In case, about 3.4:
administering to said donor an amount sufficient to enrich said donor's peripheral blood with CD34+ cells to white blood cells in a ratio of 1 to about 6.9:1. 15. The method of claim 14, wherein CD34+ to leukocytes is in a ratio of about 4.0:1 to about 6.0:1.
The donor's peripheral blood is enriched with cells. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
a CXCR2 agonist and a CXCR4 antagonist to produce a population of cells in which the ratio of CD34+ cells to neutrophils in a sample of peripheral blood of said donor is from about 0.0018 to about 0.0058 after administration of said CXCR2 agonist and CXCR4 antagonist; A method comprising administering to said donor in a sufficient amount. 17. The method of claim 16, wherein the ratio of CD34+ cells to neutrophils in the sample is about 0.0026 to about 0.0046. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
CXCR2 agonists and CXCR4 antagonists were evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and CXCR4 method. If approximately 2.1:
administering to said donor an amount sufficient to enrich said donor's peripheral blood with CD34+ cells to neutrophils in a ratio of 1 to about 8.1:1. 19. The method of claim 18, wherein the ratio of CD34+ to neutrophils is from about 5.4:1 to about 7.4:1.
The donor's peripheral blood is enriched with cells. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
a CXCR2 agonist and a CXCR4 antagonist sufficient to produce a population of cells that, after administration of said CXCR2 agonist and CXCR4 antagonist, has a ratio of CD34+ cells to lymphocytes in a sample of peripheral blood of said donor from about 0.0021 to about 0.0094. administering to said donor an amount of said donor. 21. The method of claim 20, wherein the ratio of CD34+ cells to lymphocytes in the sample is about 0.0025 to about 0.0035. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
CXCR2 agonists and CXCR4 antagonists were evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and CXCR4 method. If about 4.8:
administering to said donor an amount sufficient to enrich said donor's peripheral blood with CD34+ cells to lymphocytes to a ratio of 1 to about 8.4. 23. The method of claim 22, wherein CD34 to lymphocytes is in a ratio of about 5.0:1 to about 6.5:1.
+ Enrich the donor's peripheral blood with cells. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
A CXCR2 agonist and a CXCR4 antagonist are administered such that after administration of said CXCR2 agonist and CXCR4 antagonist, the ratio of CD34+ cells to monocytes in a sample of said donor's peripheral blood is approximately 0.
administering to said donor an amount sufficient to produce a population of cells that is from .0071 to about .0174. 25. The method of claim 24, wherein the ratio of CD34+ cells to monocytes in the sample is about 0.0100 to about 0.0140. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
CXCR2 agonists and CXCR4 antagonists were evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and CXCR4 method. administering peripheral blood of the donor in an amount sufficient to enrich the peripheral blood of the donor with CD34+ cells to monocytes in a ratio of about 1.1:1 to about 2.3:1. Method. 27. The method of claim 26, wherein the donor's peripheral blood is enriched with CD34+ cells to monocytes to a ratio of about 1.3:1 to about 1.9:1. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
After administration of the CXCR2 agonist and CXCR4 antagonist, the frequency of CD34+ cells in a sample of peripheral blood of the donor is between about 0.051% and about 0.1%.
administering to said donor an amount sufficient to produce a population of cells that is 40%. 29. The method of claim 28, wherein the frequency of CD34+ cells in the sample is between about 0.080% and about 0.
.120%. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
CXCR2 agonists and CXCR4 antagonists were evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and CXCR4 method. administering to said donor an amount sufficient to increase the frequency of CD34+ cells in said donor's peripheral blood by about 3.4-fold to about 7.1-fold when the donor has a CD34+ cell frequency. 31. The method of claim 30, wherein the frequency of CD34+ cells in peripheral blood of the donor increases from about 4.0-fold to about 6.0-fold after administration of the CXCR2 agonist and CXCR4 antagonist. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
A CXCR2 agonist and a CXCR4 antagonist are administered to CD34+ CD90+ CD45RA on leukocytes in a sample of peripheral blood of said donor after administration of said CXCR2 agonist and CXCR4 antagonist.
- administering to said donor an amount sufficient to produce a population of cells in which the proportion of cells is from about 0.0003 to about 0.0016. 33. The method of claim 32, wherein CD34+CD90+CD45 on leukocytes in the sample.
The ratio of RA- cells is about 0.0006 to about 0.0012. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
CXCR2 agonists and CXCR4 antagonists were evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and CXCR4 method. In case, about 5.5:
administering to said donor an amount sufficient to enrich peripheral blood of said donor with CD34+ CD90+ CD45RA- cells to white blood cells in a ratio of 1 to about 26.9:1. 35. The method of claim 34, wherein CD34+ to leukocytes is in a ratio of about 5.5:1 to about 6.5:1.
The donor's peripheral blood is enriched with CD90+ CD45RA- cells. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
A CXCR2 agonist and a CXCR4 antagonist are administered to CD34+ CD90+ CD45RA on neutrophils in a sample of peripheral blood of said donor after administration of said CXCR2 agonist and CXCR4 antagonist.
- administering to said donor an amount sufficient to produce a population of cells having a ratio of about 0.0007 to about 0.0043 cells. 37. The method of claim 36, wherein CD34+CD90+CD45 on neutrophils in the sample.
The proportion of RA- cells is about 0.0014 to about 0.0034. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
CXCR2 agonists and CXCR4 antagonists were evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and CXCR4 method. In case, about 3.5:
administering to said donor an amount sufficient to enrich peripheral blood of said donor with CD34+ CD90+ CD45RA- cells to neutrophils in a ratio of 1 to about 22.0:1. 39. The method of claim 38, wherein CD34+ to neutrophils is in a ratio of about 7.0:1 to about 9.0:1.
The donor's peripheral blood is enriched with CD90+ CD45RA- cells. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
A CXCR2 agonist and a CXCR4 antagonist are administered to CD34+ CD90+ CD45 lymphocytes in a sample of peripheral blood of said donor after administration of said CXCR2 agonist and CXCR4 antagonist.
A method comprising administering to said donor an amount sufficient to produce a population of cells having a ratio of RA- cells from about 0.0008 to about 0.0069. 41. The method of claim 40, wherein CD34+ CD90+ CD on lymphocytes in the sample.
The proportion of 45RA- cells is about 0.0011 to about 0.0031. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
CXCR2 agonists and CXCR4 antagonists were evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and CXCR4 method. In case, about 5.6:
administering to said donor an amount sufficient to enrich peripheral blood of said donor with CD34+ CD90+ CD45RA- cells to lymphocytes at a ratio of 1 to about 37.0:1. 43. The method of claim 42, wherein CD3 to lymphocytes is in a ratio of about 8.0:1 to about 10.0:1.
The donor's peripheral blood is enriched with 4+ CD90+ CD45RA- cells. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
A CXCR2 agonist and a CXCR4 antagonist are administered to CD34+ CD90+ CD45RA- monocytes in a peripheral blood sample of said donor after administration of said CXCR2 agonist and CXCR4 antagonist.
administering to said donor an amount sufficient to produce a population of cells having a cell ratio of about 0.0028 to about 0.0130. 45. The method of claim 44, wherein CD34+CD90+CD45RA on monocytes in the sample.
- The cell ratio is about 0.0063 to about 0.0083. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
CXCR2 agonists and CXCR4 antagonists were evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and CXCR4 method. In case, about 1.5:
administering to said donor an amount sufficient to enrich peripheral blood of said donor with CD34+ CD90+ CD45RA- cells to monocytes in a ratio of 1 to about 8.5:1. 47. The method of claim 46, wherein CD34+ CD to monocytes is in a ratio of about 1.3:1 to about 2.5:1.
The donor's peripheral blood is enriched with 90+ CD45RA- cells. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
A CXCR2 agonist and a CXCR4 antagonist are administered to CD34+ CD90+ CD4 cells in a peripheral blood sample of said donor after administration of said CXCR2 agonist and CXCR4 antagonist.
5RA- in an amount sufficient to produce a population of cells having a ratio of about 0.393 to about 0.745 to said donor. 49. The method of claim 48, wherein CD34+ CD90+ C on CD34+ cells in the sample
The ratio of D45RA- cells is about 0.625 to about 0.725. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
CXCR2 agonists and CXCR4 antagonists were evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and CXCR4 method. If approximately 1.1:
administering to said donor an amount sufficient to enrich peripheral blood of said donor with CD34+ CD90+ CD45RA- cells to CD34+ cells in a ratio of 1 to about 4.8:1. 51. The method of claim 50, wherein CD3 to CD34+ cells are in a ratio of about 1.1:1 to about 1.5:1.
The donor's peripheral blood is enriched with 4+ CD90+ CD45RA- cells. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
a CXCR2 agonist and a CXCR4 antagonist to produce a population of cells in which the frequency of CD34+ CD90+ CD45RA- cells in a sample of peripheral blood of said donor is about 0.020% to about 0.110% after administration of said CXCR2 agonist and CXCR4 antagonist; administering to said donor in an amount sufficient to. 53. The method of claim 52, wherein the frequency of CD34+ CD90+ CD45RA- cells in the sample is about 0.046% to about 0.086%. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
CXCR2 agonists and CXCR4 antagonists were evaluated by comparing a sample of the donor's peripheral blood after administration of the CXCR2 agonist and CXCR4 antagonist to a sample of the donor's peripheral blood before administration of the CXCR2 agonist and CXCR4 method. administering to said donor an amount sufficient to increase the frequency of CD34+ CD90+ CD45RA- cells in said donor's peripheral blood by about 5.1-fold to about 25.7-fold, if applicable. 55. The method of claim 54, wherein the frequency of CD34+ CD90+ CD45RA- cells in peripheral blood of the donor increases from about 5.1-fold to about 7.1-fold after administration of the CXCR2 agonist and CXCR4 antagonist. A method for mobilizing a population of hematopoietic stem cells from the bone marrow of a mammalian donor into peripheral blood, the method comprising:
The method includes:
a. administering to said donor a mobilizing amount of a CXCR2 agonist and a CXCR4 antagonist;
b. obtaining input values for each of the one or more parameters listed in Table 2 characterizing the donor peripheral blood sample; and
c. for expanding hematopoietic stem cells ex vivo, or in a mammalian patient, if the input values for each of said one or more parameters meet the corresponding reference standard for each of said one or more parameters. For use in treating stem cell disorders including:
providing said sample; 57. The method of claim 56, wherein the one or more reference parameters are any one of Tables 3-6.
This is a combination of the parameters listed in . 58. The method of claim 56 or 57, further comprising expanding the hematopoietic stem cells ex vivo when the sample is provided for expansion ex vivo. If said sample is provided for use in treating one or more stem cell disorders, the method further comprises injecting said hematopoietic stem cells or their progeny into a mammal suffering from said one or more stem cell disorders. 59. A method according to any one of claims 56 to 58, comprising: 60. The method of any one of claims 12-59, wherein the sample is isolated from the donor about 3 hours to about 5 hours after administration of the CXCR2 agonist and CXCR4 antagonist. 61. The method of claim 60, wherein the sample is isolated from the donor about 4 hours after administration of the CXCR2 agonist and CXCR4 antagonist. 62. The method of any one of claims 12-61, wherein the CXCR2 agonist is Gro-
β T or a variant thereof. 63. The method of claim 62, wherein the CXCR2 agonist is SEQ ID NO: 2
A peptide having at least 85% sequence identity to the amino acid sequence of 64. The method of claim 63, wherein the amino acid sequence of the CXCR2 agonist differs from the amino acid sequence of SEQ ID NO: 2 only by one or more conservative amino acid substitutions. 64. The method of claim 63, wherein the CXCR2 agonist is Gro-βT. 62. The method of any one of claims 12-61, wherein the CXCR2 agonist is Gro-
β or a variant thereof. 67. The method of claim 66, wherein the CXCR2 agonist is SEQ ID NO: 1
A peptide having at least 85% sequence identity to the amino acid sequence of 68. The method of claim 67, wherein the amino acid sequence of the CXCR2 agonist differs from the amino acid sequence of SEQ ID NO: 1 only by one or more conservative amino acid substitutions. 68. The method of claim 67, wherein the CXCR2 agonist is Gro-β. 70. The method of any one of claims 12-69, wherein the CXCR2 agonist is
A dose of μg/kg to about 1 mg/kg is administered to the donor. 71. The method of claim 70, wherein the CXCR2 agonist is administered at a concentration of about 50 μg/kg to about 300 μg/kg.
kg dose to said donor. 72. The method of claim 71, wherein the CXCR2 agonist is comprised between about 100 μg/kg and about 250 μg.
/kg to said donor. 73. The method of claim 72, wherein the CXCR2 agonist is administered to the donor at a dose of about 150 μg/kg. 74. The method of any one of claims 1-73, wherein said CXCR2 agonist is administered intravenously to said donor. 75. The method of any one of claims 12-74, wherein said CXCR2 agonist and CXCR4
An antagonist is simultaneously administered to said donor. 76. The method of any one of claims 12-75, wherein the CXCR4 antagonist is
Formula (I)
Z -Linker- Z' (I)
or a pharmaceutically acceptable salt thereof, where Z is:
(i) is a cyclic polyamine containing from 9 to 32 ring members, where 2 to 8 ring members are
are nitrogen atoms separated from each other by two or more carbon atoms; or
(ii) An amine represented by formula (IA)
where A comprises a monocyclic or bicyclic fused ring system containing at least one nitrogen atom, and B is H or a substituent consisting of 1 to 20 atoms;
and where Z' is:
(i) is a cyclic polyamine containing from 9 to 32 ring members, where 2 to 8 ring members are
are nitrogen atoms separated from each other by two or more carbon atoms;
(ii) An amine represented by formula (IB)
where A' comprises a monocyclic or bicyclic fused ring system containing at least one nitrogen atom, and B' is H or a substituent consisting of 1 to 20 atoms; or
(iii) Substituent represented by formula (IC)
-N(R)-(CR ₂ ) _n -X (IC)
where each R is independently H or _C1 - _C6 alkyl, n is 1 or 2, and X is an aryl or heteroaryl group or a mercaptan;
wherein said linker is a bond, an optionally substituted C ₁ -C ₆ alkylene, an optionally substituted C ₁ -C ₆ heteroalkylene, an optionally substituted C ₂ -C ₆ alkenylene, optionally substituted _C2 - _C6 heteroalkenylene, optionally substituted _C2 - _C6 alkynylene, optionally substituted _C2 - _C6 heteroalkynylene, optionally substituted optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene, or optionally substituted heteroarylene. 77. The method of claim 76, wherein Z and Z' are each independently a cyclic polyamine containing from 9 to 32 ring members, and from 2 to 8 of the 9 to 32 ring members. Individuals are nitrogen atoms separated from each other by two or more carbon atoms. 78. The method of claim 76 or 77, wherein Z and Z' are the same substituent. A method according to any one of claims 76 to 78, wherein Z is a cyclic polyamine containing 10 to 24 ring members. 80. The method of claim 79, wherein Z is a cyclic polyamine containing 14 ring members. 81. A method according to any one of claims 76 to 80, wherein Z comprises 4 nitrogen atoms. A method according to any one of claims 76 to 81, wherein Z is 1,4,8,11-tetraazocyclotetradecane. 83. The method of any one of claims 76-82, wherein the linker is represented by the formula (ID):

where Ring D is an optionally substituted aryl group, an optionally substituted heteroaryl group, an optionally substituted cycloalkyl group, or an optionally substituted heterocycloalkyl group is; and
X and Y are each independently optionally substituted _C1 - _C6 alkylene, optionally substituted _C1 - _C6 heteroalkylene, optionally substituted _C2 -C ₆ alkenylene, optionally substituted _C2 - _C6 heteroalkenylene, optionally substituted _C2 - _C6 alkynylene, or optionally substituted _C2 - _C6 heteroalkynylene . 84. The method of claim 83, wherein the linker has the formula (IE):

where Ring D is an optionally substituted aryl group, an optionally substituted heteroaryl group, an optionally substituted cycloalkyl group, or an optionally substituted heterocycloalkyl group is; and
X and Y are each independently optionally substituted _C1 - _C6 alkylene, optionally substituted _C1 - _C6 heteroalkylene, optionally substituted _C2 -C ₆ alkenylene, optionally substituted _C2 - _C6 heteroalkenylene, optionally substituted _C2 - _C6 alkynylene, or optionally substituted _C2 - _C6 heteroalkynylene . 85. The method of claim 83 or 84, wherein X and Y are each independently optionally substituted _C1 - _C6 alkylene. A method according to any one of claims 83 to 85, wherein X and Y are the same substituent. 87. The method of claim 86, wherein X and Y are each a methylene group. 88. The method of any one of claims 76-87, wherein the CXCR4 antagonist is
Plerixafor or a pharmaceutically acceptable salt thereof. 89. The method of claim 88, wherein said plerixafor or a pharmaceutically acceptable salt thereof is administered subcutaneously to said donor. 90. The method of claim 88 or 89, wherein said plerixafor, or a pharmaceutically acceptable salt thereof, is administered to said donor at a dose of about 50 μg/kg to about 500 μg/kg. 91. The method of claim 90, wherein said plerixafor, or a pharmaceutically acceptable salt thereof, is administered to said donor at a dose of about 200 μg/kg to about 300 μg/kg. 92. The method of claim 91, wherein said plerixafor, or a pharmaceutically acceptable salt thereof, is administered to said donor at a dose of about 240 μg/kg. A pharmaceutical composition comprising a population of hematopoietic stem cells or progeny thereof isolated from a mammalian donor, wherein the ratio of CD34+ cells to white blood cells in the population is from about 0.0008 to about 0.0021. 94. The pharmaceutical composition of claim 93, wherein the ratio of CD34+ cells to white blood cells in the population is about 0.0010 to about 0.0018. A pharmaceutical composition comprising a population of hematopoietic stem cells or progeny thereof isolated from a mammalian donor, wherein the ratio of CD34+ cells to neutrophils in the population is from about 0.0018 to about 0.0058. 96. The pharmaceutical composition of claim 95, wherein the ratio of CD34+ cells to neutrophils in the population is about 0.0026 to about 0.0046. A pharmaceutical composition comprising a population of hematopoietic stem cells or progeny thereof isolated from a mammalian donor, wherein the ratio of CD34+ cells to lymphocytes in the population is from about 0.0021 to about 0.0094. 98. The pharmaceutical composition of claim 97, wherein the ratio of CD34+ cells to lymphocytes in the population is about 0.0025 to about 0.0035. A pharmaceutical composition comprising a population of hematopoietic stem cells or progeny thereof isolated from a mammalian donor, wherein the ratio of CD34+ cells to monocytes in the population is from about 0.0071 to about 0.0174. 100. The pharmaceutical composition of claim 99, wherein the ratio of CD34+ cells to monocytes in the population is about 0.0100 to about 0.0140. A pharmaceutical composition comprising a population of hematopoietic stem cells or progeny thereof isolated from a mammalian donor, wherein the frequency of CD34+ cells in said population is from about 0.051% to about 0.140%. 102. The pharmaceutical composition of claim 101, wherein the frequency of CD34+ cells in the population is about 0.080%.
~0.120%. A pharmaceutical composition comprising a population of hematopoietic stem cells or progeny thereof isolated from a mammalian donor, wherein the ratio of CD34+ CD90+ CD45RA- cells to white blood cells in said population is from about 0.0003 to about 0.0016.
It is. 104. The pharmaceutical composition of claim 103, wherein CD34+CD90+ on leukocytes in said population.
The proportion of CD45RA- cells is about 0.0006 to about 0.0012. A pharmaceutical composition comprising a population of hematopoietic stem cells or their progeny isolated from a mammalian donor, wherein the ratio of CD34+ CD90+ CD45RA- cells to neutrophils in said population is from about 0.0007 to about 0.0043.
It is. 106. The pharmaceutical composition of claim 105, wherein CD34+CD90+ on neutrophils in said population.
The proportion of CD45RA- cells is about 0.0014 to about 0.0034. A pharmaceutical composition comprising a population of hematopoietic stem cells or their progeny isolated from a mammalian donor, wherein the ratio of CD34+ CD90+ CD45RA- cells to lymphocytes in said population is from about 0.0008 to about 0.00.
It is 69. 108. The pharmaceutical composition of claim 107, wherein CD34+CD90 on lymphocytes in said population.
The proportion of +CD45RA- cells is about 0.0011 to about 0.0031. A pharmaceutical composition comprising a population of hematopoietic stem cells or progeny thereof isolated from a mammalian donor, wherein the ratio of CD34+ CD90+ CD45RA- cells to monocytes in said population is from about 0.0028 to about 0.0130. 110. The pharmaceutical composition of claim 109, wherein CD34+ CD90+ CD on monocytes in said population.
The proportion of 45RA- cells is about 0.0063 to about 0.0083. A pharmaceutical composition comprising a population of hematopoietic stem cells or progeny thereof isolated from a mammalian donor, wherein the ratio of CD34+ CD90+ CD45RA- cells to CD34+ cells in said population is from about 0.393 to about 0.74.
It is 5. 112. The pharmaceutical composition of claim 111, wherein CD34+ CD9 to CD34+ cells in said population
The proportion of 0+ CD45RA- cells is about 0.625 to about 0.725. A pharmaceutical composition comprising a population of hematopoietic stem cells or their progeny isolated from a mammalian donor, wherein the frequency of CD34+CD90+CD45RA- cells in said population is about 0.020% to about 0.110%. 114. The pharmaceutical composition of claim 113, wherein the frequency of CD34+ CD90+ CD45RA- cells in the population is about 0.046% to about 0.086%. A method of treating a stem cell disorder in a mammalian patient, the method comprising:
a. mobilizing a population of hematopoietic stem cells in a mammalian donor according to the method of any one of claims 1-92; and
b. Infusing a therapeutically effective amount of hematopoietic stem cells or their progeny into said patient. 93. A method of treating a stem cell disorder in a mammalian patient, the method comprising infusing said patient with a therapeutically effective amount of hematopoietic stem cells or their progeny, mobilized by the method of any one of claims 1-92. A method including: 115. A method of treating a stem cell disorder in a mammalian patient, the method comprising administering to said patient a pharmaceutical composition according to any one of claims 93-114. 118. The method of any one of claims 115-117, wherein the stem cell disorder is a hemoglobinopathic disorder. 119. The method of claim 118, wherein the hemoglobinopathic disorder is selected from the group consisting of sickle cell anemia, thalassemia, Fanconi anemia, aplastic anemia, and Wiskott-Aldrich syndrome. 118. The method of any one of claims 115-117, wherein the stem cell disorder is a myelodysplastic disorder. 118. The method of any one of claims 115-117, wherein said stem cell disorder is an immunodeficiency disorder. 122. The method of claim 121, wherein the immunodeficiency disorder is a congenital immunodeficiency disorder. 122. The method of claim 121, wherein the immunodeficiency disorder is an acquired immunodeficiency disorder. 124. The method of claim 123, wherein the acquired immunodeficiency is human immunodeficiency virus or acquired immunodeficiency syndrome. 118. The method of any one of claims 115-117, wherein said stem cell disorder is a metabolic disorder. 126. The method of claim 125, wherein the metabolic disorder is selected from the group consisting of glycogen storage disease, mucopolysaccharidosis, Gaucher disease, Hurler disease, sphingolipid dosage, and metachromatic leukodystrophy. 118. The method of any one of claims 115-117, wherein said stem cell disorder is cancer. 128. The method of claim 127, wherein the cancer is leukemia, lymphoma, multiple myeloma,
and neuroblastoma. 124. The method of claim 123, wherein the cancer is a hematological cancer. 128. The method of claim 127, wherein the cancer is acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, multiple myeloma, diffuse large B-cell lymphoma. , or non-Hodgkin's lymphoma. 118. The method of any one of claims 115-117, wherein the stem cell disorder is adenosine deaminase deficiency and severe combined immunodeficiency, hyperimmunoglobulin M syndrome, Chediak-Higashi disease, hereditary lymphoma. histiocytosis, osteopetrosis, osteogenesis imperfecta, storage diseases, thalassemia major, systemic sclerosis, systemic lupus erythematosus
s), multiple sclerosis, and juvenile rheumatoid arthritis. 118. The method of any one of claims 115-117, wherein said stem cell disorder is an autoimmune disorder. 133. The method of claim 132, wherein the autoimmune disorder is multiple sclerosis, human systemic lupus erythematosus, rheumatoid arthritis, inflammatory bowel disease, treatment of psoriasis, type 1 diabetes, acute disseminated encephalomyelitis, Addison's disease, generalized alopecia, ankylosing spondylitis, antiphospholipid antibody syndrome, aplastic anemia, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome, Autoimmune oophoritis, Barrow disease, Behcet's disease, bullous pemphigoid, cardiomyopathy, Chagas disease, chronic fatigue immunodeficiency syndrome, chronic inflammatory demyelinating polyneuropathy, Crohn's disease, cicatricial pemphigoid, celiac disease. coeliac sprue-dermatiti
s herpetiformis), cold agglutinin disease, CREST syndrome, Degos disease, discoid lupus erythematosus,
Autonomic neuropathy, endometriosis, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, Goodpasture syndrome, Graves' disease, Guillain-Barre syndrome, Hashimoto's thyroiditis, hidradenitis suppurativa, idiopathic and /or acute thrombocytopenic purpura, idiopathic pulmonary fibrosis, IgA neuropathy, interstitial cystitis, juvenile arthritis, Kawasaki disease, lichen planus, Lyme disease, Meniere's disease, mixed connective tissue disease, myasthenia gravis neuromyotonia, opsoclonus-myoclonus syndrome, optic neuritis, ord's thyroiditis, pemphigus vulgaris, pernicious anemia,
Polychondritis, polymyositis and dermatomyositis, primary biliary cirrhosis, polyarteritis nodosa, polyendocrine syndrome, polymyalgia rheumatica, primary agammaglobulinemia
bulinemia), Raynaud's phenomenon, Reiter's syndrome, rheumatic fever, sarcoidosis, scleroderma,
selected from the group consisting of Sjögren's syndrome, stiff person syndrome, Takayasu's arteritis, temporal arteritis, ulcerative colitis, uveitis, vasculitis, vitiligo, vulvodynia, and Wegener's granulomatosis. 134. The method of any one of claims 115-133, wherein the hematopoietic stem cells are autologous to the patient. 134. The method of any one of claims 115-133, wherein the hematopoietic stem cells are allogeneic to the patient. 136. The method of claim 135, wherein the hematopoietic stem cells are HLA matched to the patient. 137. The method of any one of claims 115-136, wherein the hematopoietic stem cells are genetically modified to disrupt endogenous genes. 138. The method of claim 137, wherein the endogenous gene encodes a major histocompatibility complex protein. 139. The method of any one of claims 115-138, wherein the hematopoietic stem cells or their progeny are injected into the patient at least 2 days after the hematopoietic stem cells or their progeny are injected into the patient. maintains potential. 140. The method of any one of claims 115-139, wherein the hematopoietic stem cells or their progeny are localized in hematopoietic tissue after infusion of the hematopoietic stem cells or their progeny into the patient, and/or Reestablish hematopoiesis. 141. The method of any one of claims 115-140, wherein the hematopoietic stem cells or their progeny, when injected into the patient, contain megakaryocytes, thrombocytes, platelets, red blood cells, mast cells, Recovering a population of cells selected from the group consisting of myoblasts, basophils, neutrophils, eosinophils, microglia, granulocytes, monocytes, osteoclasts, and antigen presenting cells. 93. The method of any one of claims 1-92, wherein said mammalian donor is a human donor. A pharmaceutical composition according to any one of claims 93 to 114, wherein said mammalian donor is a human donor. 142. The method of any one of claims 115-141, wherein the mammalian donor is a human donor and the mammalian patient is a human patient. 7. The method of any one of the preceding claims, further comprising isolating the hematopoietic stem cells or their progeny by collecting peripheral blood from the donor. 7. The method of any one of the preceding claims, further comprising collecting the hematopoietic stem cells or their progeny from the donor using apheresis. The method of any one of the preceding claims, wherein the Gro-β, Gro-β T and variants thereof have a purity of at least about 95% relative to deamidated versions of these peptides. . An enriched preparation of human blood cells comprising a population of hematopoietic stem cells or progeny thereof, wherein the ratio of CD34+ cells to white blood cells in said preparation is from about 0.0008 to about 0.0021. An enriched preparation of human blood cells comprising a population of hematopoietic stem cells or progeny thereof, wherein the ratio of CD34+ CD90+ CD45RA- cells to neutrophils in said preparation is from about 0.0007 to about 0.0043. A human blood cell preparation comprising hematopoietic stem cells or their progeny, prepared using the method of any one of claims 1-92. A method of mobilizing CD34 ^dim cells from the bone marrow of a human donor into peripheral blood, the method comprising:
(i) a CXCR2 agonist selected from the group consisting of Gro-β, Gro-β T, and variants thereof at a dose of about 50 μg/kg to about 1000 μg/kg; and (ii) a CXCR4 antagonist to said donor. A method comprising administering to A method of performing an allogeneic hematopoietic stem cell transplant in a patient in need thereof, the method comprising injecting into the patient a therapeutically effective amount of allogeneic hematopoietic stem cells, the hematopoietic stem cells comprising: (i) a CXCR2 agonist selected from the group consisting of Gro-β, Gro-β T, and variants thereof at a dose of about 50 μg/kg to about 1000 μg/kg; and (ii) a CXCR4 antagonist to said donor. mobilizing from the bone marrow of a human donor into the peripheral blood of a human donor by a method comprising administering to a human donor. A method of preventing, reducing the risk of developing, or reducing the severity of post-transplant infection in a patient in need thereof, the method comprising: infusing said patient with a therapeutically effective amount of hematopoietic stem cells; Here, the hematopoietic stem cells are treated with (i) Gro-β, Gr at a dose of about 50 μg/kg to about 1000 μg/kg;
a CXCR2 agonist selected from the group consisting of o-β T, and variants thereof; and (ii
) A CXCR4 antagonist is mobilized from the bone marrow of a human donor into the peripheral blood of a human donor by a method comprising administering to said donor a CXCR4 antagonist. A method of preventing, reducing the risk of developing, or reducing the severity of graft-versus-host disease (GVHD) in a patient in need thereof, the method comprising infusing said patient with a therapeutically effective amount of hematopoietic stem cells. wherein the hematopoietic stem cells are treated with (i) a dose of about 50 μg/kg to about 1000 μg/kg;
by a method comprising administering to said donor a CXCR2 agonist selected from the group consisting of Gro-β, Gro-β T, and variants thereof, and (ii) a CXCR4 antagonist;
A method of mobilizing human donor bone marrow into human donor peripheral blood. 155. The method of any one of claims 151-154, wherein the CD34 ^dim cells are in an amount at least 2-10 times higher than when the hematopoietic stem cells are mobilized using the CXCR4 antagonist alone. present in peripheral blood. 156. The method of any one of claims 151-155, wherein the CD34 ^dim cells are capable of inhibiting the proliferation of alloreactive T-lymphocytes when administered to a recipient. 157. The method of any one of claims 151-156, wherein the CXCR2 agonist is G
ro-βT. 158. The method of any one of claims 151-157, wherein the CXCR2 agonist comprises:
A dose of about 100 μg/kg to about 250 μg/kg is administered to the donor. 159. The method of claim 158, wherein the CXCR2 agonist is administered at a concentration of about 125 μg/kg to about 225 μg/kg.
A dose of μg/kg is administered to said donor. 160. The method of claim 159, wherein the CXCR2 agonist is administered to the donor at a dose of about 150 μg/kg. 161. The method of any one of claims 151-160, wherein the CXCR2 agonist comprises:
Administer intravenously to said donor. 162. The method of any one of claims 151-161, wherein said CXCR4 antagonist is administered subcutaneously to said donor. 163. The method of any one of claims 151-162, wherein the CXCR4 antagonist is plerixafor or a pharmaceutically acceptable salt thereof. 164. The method of claim 163, wherein said plerixafor, or a pharmaceutically acceptable salt thereof, is administered to said donor at a dose of about 50 μg/kg to about 500 μg/kg. 165. The method of claim 164, wherein said plerixafor, or a pharmaceutically acceptable salt thereof, is administered to said donor at a dose of about 200 μg/kg to about 300 μg/kg. 166. The method of claim 165, wherein said plerixafor, or a pharmaceutically acceptable salt thereof, is administered to said donor at a dose of about 240 μg/kg. A population of CD34 ^dim cells derived from the method of any one of claims 151-165.