WO2002058723A2 - Chimiokines utilisees comme adjuvants de la reponse immunitaire - Google Patents

Chimiokines utilisees comme adjuvants de la reponse immunitaire Download PDF

Info

Publication number
WO2002058723A2
WO2002058723A2 PCT/US2002/001849 US0201849W WO02058723A2 WO 2002058723 A2 WO2002058723 A2 WO 2002058723A2 US 0201849 W US0201849 W US 0201849W WO 02058723 A2 WO02058723 A2 WO 02058723A2
Authority
WO
WIPO (PCT)
Prior art keywords
antigen
mage
chemokine
mip
tumor
Prior art date
Application number
PCT/US2002/001849
Other languages
English (en)
Other versions
WO2002058723A3 (fr
Inventor
Alain P. Vicari
Christophe Caux
Drake Laface
Original Assignee
Schering Corporation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US09/768,917 external-priority patent/US7217700B2/en
Application filed by Schering Corporation filed Critical Schering Corporation
Priority to EP02705895A priority Critical patent/EP1392351A2/fr
Priority to MXPA03006587A priority patent/MXPA03006587A/es
Priority to CA002434320A priority patent/CA2434320A1/fr
Priority to JP2002559057A priority patent/JP2004520043A/ja
Publication of WO2002058723A2 publication Critical patent/WO2002058723A2/fr
Publication of WO2002058723A3 publication Critical patent/WO2002058723A3/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001169—Tumor associated carbohydrates
    • A61K39/00117—Mucins, e.g. MUC-1
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • A61K38/193—Colony stimulating factors [CSF]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • A61K38/195—Chemokines, e.g. RANTES
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • A61K38/20—Interleukins [IL]
    • A61K38/2026—IL-4
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001148—Regulators of development
    • A61K39/00115—Apoptosis related proteins, e.g. survivin or livin
    • A61K39/001151—Apoptosis related proteins, e.g. survivin or livin p53
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001154—Enzymes
    • A61K39/001156—Tyrosinase and tyrosinase related proteinases [TRP-1 or TRP-2]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/00118—Cancer antigens from embryonic or fetal origin
    • A61K39/001182—Carcinoembryonic antigen [CEA]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001184—Cancer testis antigens, e.g. SSX, BAGE, GAGE or SAGE
    • A61K39/001186—MAGE
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001184—Cancer testis antigens, e.g. SSX, BAGE, GAGE or SAGE
    • A61K39/001188—NY-ESO
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/00119—Melanoma antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/00119—Melanoma antigens
    • A61K39/001191—Melan-A/MART
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/00119—Melanoma antigens
    • A61K39/001192—Glycoprotein 100 [Gp100]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001193—Prostate associated antigens e.g. Prostate stem cell antigen [PSCA]; Prostate carcinoma tumor antigen [PCTA]; PAP or PSGR
    • A61K39/001194—Prostate specific antigen [PSA]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/08—Drugs for disorders of the urinary system of the prostate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/10—Antimycotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/04—Antineoplastic agents specific for metastasis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55522—Cytokines; Lymphokines; Interferons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55522—Cytokines; Lymphokines; Interferons
    • A61K2039/55527—Interleukins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
    • A61K2039/6031—Proteins
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence

Definitions

  • the invention relates to the use of human chemokines in the treatment of disease states, including cancer.
  • the administered chemokines direct the migration of either all antigen-presenting dendritic cells or a specific subset of dentritic cells.
  • disease-specific antigen(s) and/or a moiety designed to activate dentritic cells is administered in conjunction with the chemokine(s).
  • DC Dendritic cells
  • DC are represented by a diverse population of morphologically similar cell types distributed widely throughout the body in a variety of lymphoid and non- lymphoid tissues (Caux, et al., 1995, Immunology Today 16:2; Steinman, 1991 , Ann.
  • DC are bone marrow-derived and migrate as precursors through blood stream to tissues, where they become resident cells such as Langerhans cells in the epidermis.
  • immature DC such as fresh
  • Langerhans cells are recruited at the site of inflammation (Kaplan, et al., 1992, J. Exp. Med. 175:1717-1728; McWilliam, et al., 1994, J. Exp. Med. 179:1331-1336) where they capture and process antigens, (Inaba, et al., 1986. J. Exp. Med. 164:605-613; Streilein, et al., 1989, J. Immunol. 143:3925-3933; Romani, et al., 1989., J. Exp. Med. 169:1169-1178; Pure, et al., 1990. J. Exp. Med. 172:1459-1469; Schuler, et al., 1985, J. Exp. Med. 161 :526-546).
  • Antigen-loaded DC migrate from the peripheral tissue via the lymphatics to the T cell rich area of lymph nodes, where the mature DC are called interdigitating cells (IDC)
  • IDC interdigitating cells
  • mature DC such as IDC of lymphoid organs are poor in antigen capture and processing but markedly efficient in naive T cell priming (Inaba, et al., 1986. J. Exp. Med. 164:605-613; Streilein, et al., 1989, J. Immunol. 143:3925-3933; Romani, et al., 1989, J. Exp. Med. 169:1169-1178; Pure, et al., 1990, J. Exp. Med. 172:1459-1469; Sallusto, et al., 1995, J. Exp. Med. 182:389-400; Cella, et al., 1997, Current Opin. Immunol. 9:10-16).
  • TNF ⁇ and LPS Signals provided by TNF ⁇ and LPS are known to induce in vivo migration of resident DC from the tissues to the draining lymphoid organs (De Smedt, et al., 1996, J. Exp. Med. 184:1413-1424; MacPherson, et al., 1995, J. Immunol. 154:1317-1322; Roake, et al., 1995, J. Exp. Med. 181 :2237-2247; Cumberbatch et al., 1992, Immunology. 75:257-263; Cumberbatch, et al., 1995, Immunology. 84:31- 35).
  • Chemokines are small molecular weight proteins that regulate leukocyte migration and activation (Oppenheim, 1993, Adv. Exp. Med. Biol. 351 :183-186; Schall, et al., 1994, Curr. Opin. Immunol. 6:865-873; Rollins, 1997, Blood 90:909- 928; Baggiolini, et al., 1994, Adv. Immunol. 55:97-179). They are secreted by activated leukocytes themselves, and by stromal cells including endothelial cells and epithelial cells upon inflammatory stimuli (Oppenheim, 1993, Adv. Exp. Med. Biol.
  • chemokines such as monocyte chemotactic protein (MCP)-3, MCP-4, macrophage inflammatory protein (MIP)-1 ⁇ , MIP-1 ⁇ , RANTES (regulated on activation, normal T cell expressed and secreted), SDF-1 , Teck (thymus expressed chemokine) and MDC (macrophage derived chemokine) have been reported to attract DC in vitro (Sozzani, et al., 1995, J. Immunol. 155:3292-3295; Sozzani, et al., 1997, J. Immunol. 159:1993-2000; Xu, ef al., 1996, J. Leukoc. Biol. 60:365-371 ; MacPherson, et al., 1995, J. Immunol. 154:1317-1322; Roake, ef al., 1995, J. Exp. Med. 181 :2237- 2247).
  • MCP monocyte chemotactic protein
  • mice with MHC-compatible APC were capable of rejecting a tumor challenge.
  • Studies in humans have demonstrated a similar role for DC. It has been reported that peptide-specific CTL are readily induced from purified CD8 + T cells using peptide-pulsed DC, but are not elicited when peptide-pulsed monocytes are used (Mehta-Damani et al., 1994, J. Immunology 153:996-1003).
  • Dendritic cell therapy thus represents a very promising approach to the treatment of disease, in particular, cancer.
  • the present invention fulfills the foregoing need by providing materials and methods for treating disease states by facilitating or inhibiting the migration or activation of antigen-presenting dendritic cells. It has now been discovered that chemokines are useful therapeutic agents. Disease states which can be treated in accordance with the invention include parasitic infections, bacterial infections, viral infections, fungal infections, cancer, autoimmune diseases, graft rejection and allergy.
  • the invention provides a method of treating disease states comprising administering to an individual in need thereof an amount of chemokine sufficient to increase the migration of immature dendritic cells to the site of antigen delivery.
  • a chemokine such as MCP-1 , MCP-2, MCP-3, MCP-4, MIP-1 ⁇ , MIP-3 ⁇ , RANTES, SDF-1 , Teck, DC tactin- ⁇ , 6Ckine, MDC, MIP-5 or a combination thereof is administered.
  • a disease-associated antigen such as a tumor-associated antigen is administered in conjunction with the chemokine.
  • Another aspect of the invention provides a method of treating disease states comprising administering to an individual in need thereof an amount of chemokine sufficient to decrease the migration of immature dendritic cells to the site of antigen delivery.
  • cytokines in particular GM-CSF and
  • IL-4 are administered in combination, either before or concurrently, with the chemokine.
  • Administration of GM-CSF and IL-4 stimulates generation of DC from precursors, thereby increasing the number of DC available to capture and process antigen.
  • an activating agent such as TNF- ⁇ , IFN- ⁇ RANK-L or agonists of RANK, and agonists of the toll-like receptor family of molecules is administered to provide maturation signals which drives the migration of DC from tissues toward lymphoid organs through the draining lymph.
  • the present invention also provides a method of enhancing an immune response in a mammal comprising administering chemokine MCP-4 or a biologically active fragment of MCP-4 to a mammal.
  • Human MCP-4 (hMCP-4) is active on human blood dendritic cells, recruiting dendritic cells and dendritic cell precursors from blood.
  • the chemokine is recombinant.
  • the chemokine is administered with antigen, for instance, in the form of a fusion protein of recombinant chemokine and antigen.
  • antigens can be tumor associated, bacterial, viral or fungal.
  • the present invention provides a method of enhancing an immune response in a mammal comprising administering chemokine 6Ckine or a biologically active fragment of 6Ckine to a mammal.
  • Human 6Ckine is active on human blood dendritic cells, recruiting dendritic cells and dendritic cell precursors from blood. By virtue of recruiting dendritic cells, chemokine 6Ckine acts as an anti- tumor agent, and specifically is shown to exert an angiostatic effect on tumor vasculature.
  • the chemokine is recombinant.
  • the chemokine is administered with antigen, for instance, in the form of a fusion protein of recombinant chemokine and antigen.
  • antigens can be tumor associated, bacterial, viral or fungal.
  • cytokines in particular GM-CSF and IL-4 are administered in combination, either before or concurrently with the chemokine.
  • the invention provides fusion proteins comprising MCP-4 or a biologically active portion of MCP-4 and antigen and 6Ckine or a biologically active portion of 6Ckine and antigen.
  • fusion proteins can be administered to a mammal in the form of a plasmid, viral vector or in the form of a recombinant vector.
  • Fig. 1 shows that immunization with a plasmid containing MIP-3 ⁇ and a tumor associated antigen has a protective effect against tumor engraftment.
  • Fig. 2 shows greater CTL activity with the administration of chemokine MIP- 3 ⁇ .
  • Fig. 3 shows the nucleotide and partial amino acid sequence of chemokine hMCP-4.
  • Fig. 4 shows that hMCP-4 injection promotes the recruitment of dendritic cells in vivo in the mouse in a dose-dependent manner.
  • Fig. 5 shows that hMCP-4 is active in recruiting dendritic cells in human blood.
  • Fig. 6 shows that MCP-4 injection increases the antigen-specific humoral response following beta-galactosidase DNA immunization.
  • Fig. 7 shows that MCP-4 increases the anti-tumor effect induced by beta- galactosidase DNA immunization when mice are challenged with a C26 colon carcinoma cell line that expresses beta-galactosidase.
  • Fig. 8 shows that h6Ckine is active in recruiting dendritic cells in human blood.
  • Fig. 9 shows that C26 colon carcinoma tumor cells engineered to express m6Ckine are less tumorigenic and that this effect depends on CD8+ cells and Natural Killer cell activity, in vivo.
  • Fig. 10 shows that C26 tumors expressing m ⁇ Ckine are significantly infiltrated by dendritic cells and CD8+T cells compared with parental tumors.
  • Fig. 11 shows that C26 colon carcinoma tumor cells engineered to express m6Ckine are less angiogenic than the parental C26 tumor.
  • Fig. 12 shows that injection of h6Ckine slows tumor growth in mice in vivo.
  • Fig. 13 shows that 6Ckine inhibits tumor growth and spontaneous metastasis in established tumors in vivo.
  • chemokines can be used in accordance with the invention as adjuvants to attract selectively the immature DC subsets at the site of antigen delivery.
  • the invention provides a method of blocking DC functions by interfering with their migration through e.g., the development of CCR6, CCR7, and CCR2 agonists and antagonists.
  • DC can induce tolerance.
  • DC found in the medulla of the thymus play a role in the negative selection of developing self- reactive thymocytes (Brocker, et al., 1997, J. Exp. Med. 185(3):541-550).
  • DC can also tolerize self-reactive peripheral T cells (Kurts, ef al., 1997, J. Exp. Med. 186(2):239-245; Adler, ef al., 1998, J. Exp. Med. 187(10): 1555-1564).
  • tolerizing DC autoimmunity, allergy
  • DC-2 TH1 greater that TH2, i.e., in allergy
  • a chemokine for use in the invention is a natural protein of the body that is active on a restricted subset of DC, in particular, immature DC.
  • chemokines including, but not limited to, MIP-3 ⁇ , Teck, MDC and MCP-4, and 6Ckine have been identified by the inventors.
  • the chemokine used in practicing the invention may be a recombinant protein with an amino-acid sequence identical to the natural product, or a recombinant protein derived from the natural product but including modifications that changes its pharmacokinetic properties while keeping its original chemoattractant property.
  • the mode of delivery of the chemokine may be by injection, including intradermal, intramuscular and subcutaneous, or topical, such as an ointment or a patch.
  • the chemokine may also be delivered as a nucleic acid sequence by the way of a vector, such as a viral vector (e.g., adenovirus, poxvirus, retrovirus, lentivirus), or an engineered plasmid DNA.
  • a viral vector e.g., adenovirus, poxvirus, retrovirus, lentivirus
  • chemokine as used herein includes chemotactic agents.
  • a chemotactic agent may be a small chemical compound which is a selective agonist of a chemokine receptor expressed by immature DC.
  • CCR6 the natural receptor of the chemokine MIP-3 ⁇ is an example of such a receptor.
  • the chemokine is administered with a disease-associated antigen.
  • the antigen can be any molecular moiety against which an increase or decrease in immune response is sought. This includes antigens derived from organisms known to cause diseases in man or animal such as bacteria, viruses, parasites (e.g., Leishmania) and fungi. This also includes antigens expressed by tumors (tumor-associated antigens) and plant antigens (allergens).
  • Tumor associated antigens for use in the invention include, but are not limited to Melan-A, tyrosinase, p97, ⁇ -HCG, GalNAc, MAGE-1 , MAGE-2, MAGE-3, MAGE- 4, MAGE-12, MART-1 , MUC1 , MUC2, MUC3, MUC4, MUC18, CEA, DDC, melanoma antigen gp75, HKer 8, high molecular weight melanoma antigen, K19, Tyr1 and Tyr2, members of the pMel 17 gene family, c-Met, PSA, PSM, ⁇ - fetoprotein, thyroperoxidase, gp100, NY-ESO-1 , telomerase and p53.
  • This list is not intended to be exhaustive, but merely exemplary of the types of antigen which may be used in the practice of the invention.
  • Different combinations of antigens may be used that show optimal function with different ethnic groups, sex, geographic distributions, and stage of disease.
  • at least two or more different antigens are administered in conjunction with the administration of chemokine.
  • the antigen can by delivered or administered at the same site and the same time as the chemokine, or after a delay not exceeding 48 hours.
  • Concurrent or combined administration means the chemokine and antigen are administered to the subject either (a) simultaneously in time, or (b) at different times during the course of a common treatment schedule. In the latter case, the two compounds are administered sufficiently close in time to achieve the intended effect.
  • the antigen can be in the form of a protein, or one or several peptides, or of a nucleic acid sequence included in a delivery vector.
  • Both primary and metastatic cancer can be treated in accordance with the invention.
  • Types of cancers which can be treated include but are not limited to melanoma, breast, pancreatic, colon, lung, glioma, hepatocellular, endometrial, gastric, intestinal, renal, prostate, thyroid, ovarian, testicular, liver, head and neck, colorectal, esophagus, stomach, eye, bladder, glioblastoma, and metastatic carcinomas.
  • carcinoma refers to malignancies of epithelial or endocrine tissues including respiratory system carcinomas, gastrointestinal system carcinomas, genitourinary system carcinomas, prostatic carcinomas, endocrine system carcinomas, and melanomas.
  • Metastatic as this term is used herein, is defined as the spread of tumor to a site distant to from the primary tumor including regional lymph nodes.
  • a moiety designed to activate, induce or stimulate maturity of the DC may be advantageously administered.
  • Such agents provide maturation signals which promote migration from the tissues to the lymph nodes.
  • This moiety can be a natural product of the body such as TNF- ⁇ or RP-105, or an agonist antibody recognizing a specific structure on DC such as an anti-CD-40 antibody, or another substance.
  • the activating substance can be a sequence of nucleic acids containing unmethylated CpG motifs or agonist of a toll-like receptor known to stimulate DC.
  • these nucleic acid sequences may be part of the vector.
  • GM-CSF and IL-4 can advantageous be administered in combination with the chemokine and/or antigen.
  • the administration combination of GM-CSF and IL-4 stimulates generation of DC from precursors.
  • GM-CSF and IL-4 may be administered for purposes of increasing the number of circulating immature DC which might then be locally recruited locally be the subsequent injection of chemokine(s). This protocol would imply a systemic pre-treatment for a least five to seven days with GM-CSF and IL-4.
  • An alternative would be to favor by local administration of GM-CSF and IL-4 the local differentiation of DC-precursors (monocytes) into immature DC which could then pick up the antigen delivered at the same site.
  • chemokine(s) and/or antigen(s) and/or activating agent(s) and/or cytokine(s) are administered as pharmaceutical compositions comprising an effective amount of chemokine(s) and/or antigen(s) and/or activating agent(s) and/or cytokine(s) in a pharmaceutical carrier.
  • reagents can be combined for therapeutic use with additional active or inert ingredients, e.g., in conventional pharmaceutically acceptable carriers or diluents, e.g., immunogenic adjuvants, along with physiologically innocuous stabilizers and excipients.
  • a pharmaceutical carrier can be any compatible, non-toxic substance suitable for delivering the compositions of the invention to a patient.
  • compositions for administration are discussed therein and below, e.g., for intravenous, intraperitoneal, or intramuscular administration, transdermal diffusion, and others.
  • Pharmaceutically acceptable carriers will include water, saline, buffers, and other compounds described, e.g., in the Merck Index, Merck & Co., Rahway, New Jersey. Slow release formulations, or a slow release apparatus may be used for continuous administration.
  • Dosage ranges for chemokine(s) and/or antigen(s) and/or activating agent(s) would ordinarily be expected to be in amounts lower than 1 mM concentrations, typically less than about 10 ⁇ M concentrations, usually less than about 100 nM, preferably less than about 10 pM (picomolar), and most preferably less than about 1 fM (femtomolar), with an appropriate carrier.
  • treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under the circumstance is reached. Determination of the proper dosage and administration regime for a particular situation is within the skill of the art.
  • the preferred biologically active dose of GM-CSF and IL-4 in the practice of the claimed invention is that dosing combination which will induce maximum increase in the number of circulating CD14 + /CD13 + precursor cells; the expression of antigen presenting molecules on the surface of DC precursors and mature DC; antigen presenting activity to T cells; and/or stimulation of antigen-dependent T cell response consistent with mature DC function.
  • the amount of IL-4 to be used for subcutaneously administration typically ranges from about 0.05 to about 8.0 ⁇ g/kg/day, preferably 0.25 - 6.0 ⁇ g/kg/day, most preferably 0.50 - 4.0 ⁇ g/kg/day.
  • the amount of GM-CSF is to be used for subcutaneous administration typically ranges from about 0.25 ⁇ g/kg/day to about 10.0 ⁇ g/kg/day, preferably from about 1.0 - 8.0 ⁇ g/kg/day, most preferably 2.5 - 5.0 ⁇ g/kg/day.
  • An effective amount for a particular patient can be established by measuring a significant change in one or more of the parameters indicated above.
  • chemokine MCP-4 or a biologically active fragment of MCP-4 promotes the recruitment of dendritic cells in vivo in the mouse in a dose-dependent manner and is also active on human dendritic cells isolated from blood.
  • Biologically active fragment means a portion of the MCP-4 molecule which is sufficient to stimulate a measurable immune response. This response can be measured as an enhanced antigen specific stimulation of immunoglobulin levels in serum, typically known as a B-cell response.
  • a biologically active fragment of MCP-4 will stimulate the production of certain classes of immunoglobulins such as lgG2a that require an increase in T Cells.
  • a biologically active fragment of MCP-4 will enhance an antigen-specific anti-tumor response.
  • An enhanced response could be measured by a slower tumor growth or lower tumor incidence following challenge with a tumor expressing the antigen.
  • An enhanced immune response could also be measured by analyzing the antigen- specific cytotoxic response of defined populations of lymphocytes (blood, spleen, lymph nodes, tumor).
  • small molecules that are CCR2 agonists e.g., found by drug discovery screen
  • the rationale is that all MCPs (1-4) are natural CCR2 agonists, and subsequently an artificial, small molecule agonist may have the same effect.
  • Many current therapeutics are small molecules obtained by organic chemistry synthesis.
  • Preferred embodiments consist of but are not restricted to recombinant hMCP-4 protein alone or combined with substances allowing for its slow release at delivering site (depot); fusions proteins consisting of hMCP-4 or fraction of hMCP-4 and an antigen (peptide more than 9 amino acids or protein); DNA or viral vector encoding for hMCP-4 or fraction of hMCP-4 with or without an antigen (peptide more than 9 amino acids or protein), or a nucleic acid sequence included in a delivery vector.
  • Human MCP-4 belongs to the CC family of chemokines. Its sequence was first published in 1996. (Uguccioni ef al., 1996, Monocyte
  • MCP-4 Chemotactic Protein 4
  • Human MCP-4 is a peptide of 8.6 kDa that consists of 75 amino acid residues.
  • Fig. 3. It is also known as CK- ⁇ - 10, SCY-A13 and NCC-1 (Swiss-Prot accession number Q99616) and was renamed CCL13 in the new chemokine nomenclature. (Zlotnik ef al., 2000, Chemokines: A New Classification System and Their Role In Immunity, Immunity, 12:121-127).
  • 6Ckine belongs to the CC family of chemokines (Hedrick, ef al., 1997, J. Immunol. 159: 1589-1593.) It is also known as CK- ⁇ -9, exodus-2 and SLC (Swiss- Prot accession number O00585 for human protein) and was renamed CCL21. Human 6Ckine (h6Ckine) binds to the chemokine CCR7 while mouse 6Ckine
  • 6Ckine like MIP-3 ⁇ and MCP-4 induces the migration of mature DC.
  • 6Ckine, as well as MIP-3 ⁇ can induce the migration of all human DC populations after maturation, including CD1a+ Langerhans cells, CD14+ interstitial DC, monocyte-derived DC, circulating blood CD11c+ DC, monocytes, and circulating blood CD11c- plasmacytoid DC.
  • the response to 6Ckine is observed after maturation induced by several DC activators, including CD40-L, TNF- ⁇ , and LPS.
  • CCR7 is up-regulated during DC activation, via 6Ckine, likely explaining the response to 6Ckine.
  • chemokine h6Ckine could be used in cancer treatment.
  • Preferred embodiments consist of but are not restricted to: recombinant h6Ckine protein alone or combined with substances allowing for its slow release at delivering site (depot at tumor site); fusion proteins or constructs made by chemical ligation consisting of h6Ckine or fraction of h6Ckine and a targeting moiety allowing delivery of the construct into tumors (e.g., antibody or fragment of antibody, protein ligand, peptide of more than 10 amino acids); DNA or viral vector (e.g., adenovirus) encoding for h6Ckine or fraction of h6Ckine with or without a targeting moiety as described above.
  • EXAMPLES The invention can be illustrated by way of the following non-limiting examples, which can be more easily understood by reference to the following materials and methods.
  • GM-CSF specific activity: 2.10 6 U/mg, Schering-Plough Research Institute, Kenilworth, NJ
  • Recombinant human TNF ⁇ specific activity: 2x10 7 U/mg, Genzyme, Boston, MA
  • Recombinant human SCF specific activity: 4x10 5 U/mg, R&D Abington, UK
  • Recombinant human IL-4 (specific activity: 2.10 7 U/mg, Schering-Plough Research Institute, Kenilworth, NJ) was used at a saturating concentration of 50 U/ml.
  • Recombinant human chemokines MIP-1 ⁇ (specific activity: 2x10 5 U/mg, 9x10 12 U/M), RANTES (specific activity: 1x10 4 U/mg, 8x10 10 U/M), MIP-3 (specific activity: 4x10 5 U/mg, 3x10 12 U/M) and MIP-3 ⁇ (specific activity: 1x10 4 U/mg, 9x10 10 U/M) were obtained through R&D (Abington, UK).
  • LPS was used at 10 ng/ml (Sigma).
  • the murine CD40 ligand transfected cell line (CD40-L L cells) was used as a stimulator of DC maturation.
  • CD34 + cells were seeded for expansion in 25 to 75 cm ⁇ culture vessels (Linbro, ICN Biomedicals, Acron, OH) at 2x10 4 cells/ml. Optimal conditions were maintained by splitting these cultures at day 5 and 10 with medium containing fresh GM-CSF and TNF ⁇ (cell concentration: 1-3x10 5 cells/ml). At day 12, between 70 to 90% of the cells are CD1a + DC.
  • Monocytes were purified by immunomagnetic depletion (Dynabeads, Dynal Oslo, Norway) after preparation of PBMC followed by a 52% Percoll gradient. The depletion was performed with anti- CD3 (OKT3), anti-CD19 (4G2), anti-CD8 (OKT8), anti-CD56 (NKH1 , Coulter Corporation, Hialeah, FL) and anti-CD16 (ION 16, Immunotech) monoclonal antibodies. Monocyte-derived dendritic cells were produced by culturing purified monocytes for 6-7 days in the presence of GM-CSF and IL-4 (Sallusto, ef al., 1994, J. Exp. Med. 179:1109-1118).
  • CD34 + HPC were cultured until day 6 in presence of GM-CSF+TNF ⁇ and in presence of GM-CSF alone from day 6 to day 12 in order to preserve their immaturity.
  • Immature DC from CD34 + HPC or monocyte-derived DC were activated for 3h to 72h in presence of TNF ⁇ (2.5 ng/ml) or LPS (10 ng/ml) or CD40L transfected L cells (1 L cells for 5 DC) as described (Caux, et al., 1994, J. Exp. Med. 180:1263-1272) .
  • CD11c + DC were prepared as previously described from peripheral blood or tonsils (Grouard, ef al., 1996, Nature 384:364-367). Briefly, tonsils obtained from children undergoing tonsillectomy were finely minced and digested with collagenase IV and DNase I (Sigma). The collected cells were centrifuged through Ficoll-Hypaque with SRBC (BioMerieux, Lyon, France) for 15 min at 500 rpm, then for 30 min at 2000 rpm. Peripheral blood mononuclear cells (PBMC) were isolated by Ficoll-Hypaque.
  • SRBC BioMerieux, Lyon, France
  • CD3 + T cells OKT3
  • CD19 + B cells 4G7
  • CD14 + monocytes MOP9
  • a second depletion was performed with anti-NKH1 , anti- glycophorine A (Immunotech) and anti-CD20 (1 F54).
  • CD4 + CD11c + lineage- DC were isolated by cell sorting using a FACStarPlus® (laser setting: power 250 mW, excitation wavelength 488 nm). All the procedures of depletion, staining and sorting were performed in presence of 0.5 mM EDTA. Reanalysis of the sorted population showed a purity > 97%.
  • Chemotaxis assay Cell migration was evaluated using a chemotaxis microchamber technique (48-well Boyden microchamber, Neuroprobe, Pleasanton, CA) (Bacon, ef al., 1988, Br. J. Pharmacol. 95:966-974). Briefly, human recombinant MIP-3 ⁇ and MIP-3 ⁇ , MIP-1 ⁇ and RANTES were diluted to concentrations ranging from 1 ng/ml to 1000 ng/ml in RPMI 1640 medium, and were added to the lower wells of the chemotaxis chamber.
  • a chemotaxis microchamber technique 48-well Boyden microchamber, Neuroprobe, Pleasanton, CA
  • human recombinant MIP-3 ⁇ and MIP-3 ⁇ , MIP-1 ⁇ and RANTES were diluted to concentrations ranging from 1 ng/ml to 1000 ng/ml in RPMI 1640 medium, and were added to the lower wells of the chemotaxis chamber.
  • RNA was diluted in total RNA, 25 ng/ ⁇ l oligo dTi2-18 primers (Pharmacia, Orsay, France) and the Superscript kit (Superscript II RNase H- Reverse Transcripase, Gibco BRL), as described by the manufacturer.
  • Superscript kit Superscript II RNase H- Reverse Transcripase, Gibco BRL
  • RT-PCR analysis Semi-quantitative PCR was performed in a Perkin Elmer 9600 thermal cycler, in a final volume of 100 ⁇ l reaction mixture containing 2.5 U AmpliTaq enzyme (5U/ ⁇ l, Perkin Elmer, Paris, France) with its 1X buffer, 0.2 mM of each dNTP (Perkin Elmer, Paris, France), 5% DMSO, and 1 ⁇ M of each forward and reverse primers.
  • CCR6 (Accession No. Z79784) and CCR7 (Accession No. L08176) primers were designed within regions of lowest homology between the chemokine receptors.
  • PCR products were visualized on 1.2% agarose gels containing 0.5 ⁇ g/ml ethidium bromide.
  • Reaction products migrating at the predicted size (1 ,021 bp for CCR6 and 1 ,067 bp for CCR7) were gel purified and subcloned into pCRII TA cloning vector (Invitrogen, Leek, The Netherlands) for sequencing verification on an ABI 373A Sequencer (Applied Biosystems, Foster City, CA.) using dye terminator technology.
  • AGAGGAGCAGCAGTGAGCAA -3' were used as probes for hybridization with the PCR products separated on 1.2% agarose gel and blotted onto Hybond N + membranes (Amersham, Les Ulis, France).
  • Intracellular Ca2 + concentration was measured using the fluorescent probe lndo-1 , according to the technique reported by Grynkiewicz ef al. (J. Biol. Chem., 1985, 260:3440-3450) Briefly, cells were washed in PBS and resuspended at 10 7 cells/ml in complete RPMI 1640 medium (see above). Then, cells were incubated for 45 min at room temperature with 3 ⁇ g/ml lndo-1 AM (Molecular Probes) in the dark. After incubation, cells were washed and resuspended in HBSS/1 % FCS at 10 7 cells/ml.
  • In situ hybridization In situ hybridization was performed as described (Peuchmaur, et al., 1990, Am. J. Pathol. 136:383-390). Two couple primers were used for amplifying by RT-PCR the majority of the open reading frame of M I P-3 ⁇ (Accession No. D86955) and M I P-3 ⁇ 3 ⁇ (Accession No. U77180) genes. +77/MIP-3 ⁇ 5'- TTGCTCCTGGCTGCTTTG -3' forward primer and -425/MIP-3 ⁇ 5'- ACCCTCCATGATGTGCAAG -3' reverse primer, +25/MIP-3 ⁇ 5'-
  • CACACTCACACTCACACACAC -3' reverse primer were used as described above with an annealing temperature at 62°C. Then, PCR products were cloned into pCRII TA cloning vector (Invitrogen, Leek, The Netherlands) for the generation of sense and anti-sense probes with the adapted promoters. Sense and antisense 35 S- labeled probes of MIP-3 ⁇ and MIP-3 ⁇ , were obtained by run off transcription of the 367 bp and 435 bp fragments, respectively. Six ⁇ m human tonsil sections were fixed in acetone and 4% paraformaldehyde followed by 0.1 M triethanolamine/0.25% acetic anhydride. The sections were hybridized overnight, RNAse A treated and exposed for 24 days. After development sections were stained with hematoxylin.
  • DC were generated from CD34 + HPC cultured in the presence of GM-CSF+ TNF ⁇ , and tested at different days of culture for their ability to migrate in response to chemokines in Boyden microchambers.
  • MIP-3 ⁇ and MIP-3 ⁇ recruited 2 to 3 times more CD34 + - derived DC than MIP-1 ⁇ or RANTES.
  • MIP-3 ⁇ and MIP-3 ⁇ attracted DC collected at different time points of the culture.
  • the response to MIP-3 ⁇ was already detected at day 4, maximal at day 5-6 and lasted until day 10. At day 13 to 14, the response to MIP-3 ⁇ was usually lost.
  • CD34 + -derived DC were sorted by FACS at day 10 of culture according to CD86 expression into immature DC (CD1a + CD86 " ) and mature DC (CD1a + CD86 + ).
  • CD1a + CD86 " responded exclusively to MIP-3 ⁇ while
  • CD1a + CD86 + responded mainly to MIP-3 ⁇ . These observations also confirmed that the cells recruited by MIP-3 ⁇ and MIP-3 ⁇ were indeed DC (CD1a + ). The correlation between DC maturation and chemokine responsiveness was further illustrated when the immaturity of DC was preserved by removing TNF ⁇ from day 6 to day 12 and when their maturation was synchronized by addition of TNF ⁇ , LPS or CD40L. Response to MIP-3 ⁇ had strongly decreased upon 48h maturation with TNF ⁇ , LPS and CD40L. Meanwhile, the response to MIP-3 ⁇ was induced by all three signals, CD40L and LPS being more potent than TNF ⁇ .
  • CCR6 mRNA During DC development from CD34 + HPC, CCR6 mRNA was first detected at day 6, increased up to day 10 after when it decreased and became barely detectable at day 14. In contrast, CCR7 mRNA appeared at day 10 and steadily increased up to day 14. Moreover, CD40L-dependent maturation induced progressive down- regulation of CCR6 mRNA which became almost undetectable after 72h, and up- regulation of CCR7 mRNA as early as 24h. Similar results were obtained after either LPS or TNF ⁇ -induced DC maturation. The up-regulation of CCR7 mRNA following activation was confirmed by Southern blot analysis of cDNA libraries.
  • MIP-3 ⁇ In line with the migration assays, and the regulation of CCR6 and CCR7 expression, MIP-3 ⁇ induced a Ca2 + flux exclusively in resting/immature DC and MIP-3 ⁇ in mature DC only. Maximal Ca2 + fluxes were observed with 30 ng/ml of MIP-3 ⁇ and 30 ng/ml of MIP-3 ⁇ , on immature and mature DC, respectively. These results show that changes in responsiveness to MIP-3 ⁇ and MIP-3 ⁇ are linked to the regulation of CCR6 and CCR7 mRNA expression, and suggest that CCR6 and CCR7 are the major functional receptors expressed on DC for MIP-3 ⁇ and MIP-3 ⁇ , respectively.
  • the response to MIP-3 ⁇ is also induced upon maturation of monocyte-derived DC
  • Monocyte-derived DC generated by culturing monocytes in presence of GM-
  • CSF+IL-4 for 6 days are typically immature DC (CD1a + , CD14", CD80 low ,
  • CD86'ow ⁇ CD83- (Cella, ef al., 1997, Current Opin. Immunol. 9:10-16; Sallusto, ef al., 1994, J. Exp. Med. 179:1109-1118). They migrated in response to MIP-1 ⁇ and RANTES but neither to MIP-3 nor to MIP-3 ⁇ . The lack of response of monocyte- derived DC to MIP-3 ⁇ is in accordance with the absence of CCR6 expression on those cells (Power, ef al., 1997, J. Exp. Med. 186:825-835; Greaves, ef al., 1997, J. Exp. Med. 186:837-844).
  • Example 4 The physiological relevance of the findings reported in Example 4 was addressed through the analysis of MIP-3 ⁇ and MIP-3 ⁇ mRNA expressions by in situ hybridization on sections of inflamed tonsils.
  • mRNA for MIP-3 ⁇ was detected at high levels in inflamed epithelial crypts but not in T cell rich areas nor in B cell follicles.
  • MIP-3 ⁇ expression was restricted to cells lining the epithelial crypts.
  • expression of MIP-3 ⁇ mRNA was restricted to T cell rich areas. The strongest signal was present in scattered cells, with a distribution overlapping that of IDC. Outside the paracortical area, no signal could be detected in B cell follicles, nor in epithelial crypts.
  • Serial sections showed clear absence of MIP-3 ⁇ expression within epithelial crypts where MIP-3 ⁇ was abundantly present. Sense probes for MIP-3 ⁇ and MIP-3 ⁇ , did not generate background hybridization.
  • MIP-3 ⁇ expression is restricted to inflamed epithelium, at the site of antigen entry where immature DC should be recruited.
  • MIP-3 ⁇ is only detected in paracortical areas, where mature IDC home and generate primary T cell responses.
  • MIP-3 ⁇ was shown by the inventors to be a chemotactic factor for mouse immature dendritic cells in vitro, the ability of the chemokine MIP-3 ⁇ to attract immature DC in vivo and to modulate the antigen-specific immune response against a tumor in vivo was studied. If a tumor-associated antigen is delivered at the same time, more DC will be available to capture the antigen, and therefore the antigen- specific response against this antigen should be increased.
  • Chemokine was delivered in vivo via a plasmid vector (pcDNA3, InVitrogen), that contains the cDNA encoding mouse MIP-3 ⁇ under the control of the CMV promoter (PMIP-3 ⁇ ).
  • the antigen used was ⁇ -galactosidase isolated from E. coli.
  • the antigen was delivered in vivo via the same plasmid vector pcDNA3 (called pLacz).
  • the tumor was a C26 colon carcinoma syngeneic in BALB/c mice that has been stably transfected with the gene encoding for ⁇ -galactosidase. Therefore, in this system, ⁇ -galactosidase defines a tumor-associated antigen.
  • mice Groups of 6 female 6 week-old mice were injected with either the empty pcDNA3 plasmid (negative control), the plasmid pLacz encoding the antigen alone, or a mixture of pLacz and PMIP-3 ⁇ . Injections (50 ⁇ g of total plasmid) were performed in the hind footpad every week for 4 weeks. After that time, mice were injected subcutaneously with the C26 tumor cell line expressing ⁇ -galactosidase. Typically, all mice develop subcutaneous tumors after 10 days. The appearance of tumors in these groups of mice were monitored. It was found that the appearance of tumors was delayed after pLacz and pLacz+PMIP- ⁇ injection. (Fig.
  • mice T cell-mediated antigen-specific cytotoxicity (CTL activity). Therefore, the CTL activity in the same groups of mice was analyzed 30 days after tumor inoculation. Spleen cells were removed and stimulated for five days with irradiated syngeneic DC plus an immunodominant CTL peptide derived from ⁇ -galactosidase in the presence of interleukin-2.
  • CTL activity T cell-mediated antigen-specific cytotoxicity
  • Chemokine hMCP-4 administration in an in vivo mouse model The inventors have shown that hMCP-4 local injection can promote the recruitment of dendritic cells in vivo in the mouse in a dose-dependent manner. (Fig. 4)
  • mice 6- to 10-week-old female BALB/c mice were purchased from Charles River (Iffa-Credo, L'Arbresle, France) and maintained in our facilities under standard conditions. Procedures involving animals and their care were conducted in conformity with EEC (European Economic Community) Council Directive 86/609, OJL 358,1 , December 12, 1987. Recombinant human MCP-4 protein, >97% pure (Fig. 3), was obtained from Peprotech and resuspended in PBS (Gibco-BRL). Groups of three mice were injected with PBS alone or varying amounts of human MCP-4 in PBS, intracutaneously in the right hind footpad under a 50 ⁇ volume.
  • EEC European Economic Community
  • mice were sacrificed after 2 or 20 hours and the skin at the site of injection as well as the popliteal lymph node, draining the injection site, removed. Local cell recruitment in the skin was examined by immuno-histochemistry with specific monoclonal antibodies according to standard techniques.
  • Cell suspensions were prepared from lymph nodes in RPMI 1640 + 10% fetal calf serum (FCS) (Gibco-BRL). Cell were numerated and stained in PBS + 2% FCS with biotin-CD11c and FITC-CD11 b antibodies (Becton Dickinson), followed by PE-streptavidin (Dako), according to standard procedures. Expression of CD11 b and CD 11c, that define populations of mouse dendritic cells, was analyzed on a Facscan flow cytometer (Becton
  • hMCP-4 induces the recruitment of dendritic cells identified by the co-expression of CD11 b and CD11c, but only after a longer period (20 hours). This delay most likely corresponds to the maturation and migration time necessary for dendritic cells or their precursors, initially recruited in the skin, in order to migrate to the draining lymph node.
  • hMCP-4 is also active on human dendritic cells, including dendritic cells isolated from blood. (Fig. 5)
  • Panel A Human circulating blood CD11c+ DC were enriched by magnetic bead depletion, and studied in transwell (5 ⁇ m pore size) migration assay, in response to various chemokines. The migration was revealed after 2 hours by triple staining: lineage markers FITC, HLA-DR tricolor, and CD11c PE, and analyzed by Facs. Each chemokine was tested over a wide range of concentrations (1 to 1000 ng/ml) and only the optimal response is shown. Results are expressed as migration index and represent the mean values obtained from 3 to 10 independent experiments. Blood CD11c+ mainly respond to MCP-4 as well as to MCP1 , 2 and 3 (not shown). SDF-1 , lacking selectivity, being the only other chemokine strongly active on CD11c+ DC.
  • Panel B Different human DC and DC precursor populations including blood CD11c+ DC, monocytes, monocyte-derived DC, CD1a+ Langerhans cell precursors and CD14+ interstitial DC precursors were studied in transwell (5 ⁇ m pore size) migration assay, in response to MCP-1 and MCP-4. All populations respond to MCP-4 except CD1a+ Langerhans cell precursors. In addition monocyte-derived DC respond to MCP-4 but not to MCP-1 , through a receptor different from CCR2.
  • MCP-4 is active on human DC.
  • MCP-4 is the most potent chemokine inducing the migration of circulating blood CD11c+ DC.
  • MCP-1 , and MCP-2 and MCP-3 display a similar activity on blood DC. The MCPs likely recruit blood DC through CCR2 which is highly expressed on these cells.
  • MCP-4 is active on all DC or DC precursors populations (blood CD11c+DC, monocytes, monocyte-derived DC, CD14+ interstitial type DC precursors) except the CD1a+ Langerhans cell precursors which do not express CCR2.
  • MCP-4, but not MCP-1 induces the migration of monocyte derived DC, likely through a receptor different from CCR2.
  • Example 9 hMCP-4 and ⁇ -galactosidase administration in an in vivo mouse model
  • hMCP-4 can be used as adjuvant of an antigen-specific immune response induced by plasmid DNA vaccination.
  • hMCP-4 when used as adjuvant of plasmid DNA vaccination, it can increase the protection of mice subsequently challenged with a tumor expressing the antigen encoded by the plasmid DNA.
  • mice Groups of seven 6 to 8 week-old female BALB/c mice (Iffa-Credo, L'Arbresle, France) were injected with PBS alone or 100 ng of human MCP-4 in PBS, intracutaneously in the right hind footpad under a 50 ⁇ l volume. After three hours, mice were injected at the same site with 50 ⁇ g of control pcDNA3 plasmid
  • Serum was collected one day before the first immunization and one week after the last immunization.
  • Levels of beta-galactosidase specific immunoglobulins in serum were measured with specific ELISA assays as previously described (Mendoza et al., 1997, J. Immunol. 159:5777-5781 ).
  • MCP-4 injection increases the antigen-specific humoral response following beta-galactosidase DNA immunization (50 micrograms DNA injection 3 hours after 100 ng hMCP-4 injection in rear right footpad).
  • Figure 6 shows anti-betagalactosidase antibodies measured after 4 immunizations [significance hMCP-4 + pLacz compared with PBS + pLacz: Student's test.]
  • mice were challenged with a subcutaneous injection in the right flank of 5 x 10 4 C26-BAG colon carcinoma cells which express beta-galactosidase (a kind gift from Mario Colombo, Instituto Nazionale Tumori, Milan, Italy), under a 100 ⁇ l volume of RPMI-1640.
  • beta-galactosidase a kind gift from Mario Colombo, Instituto Nazionale Tumori, Milan, Italy
  • MCP-4 injection increases the anti-tumor effect induced by beta-galactosidase DNA immunization (50 micrograms DNA injection 3 hours after 100 ng hMCP-4 injection in rear right footpad, four immunizations prior to tumor challenge) when mice are challenged with a C26 colon carcinoma cell line that expresses beta-galactosidase [significance hMCP-4 + pLacz compared with PBS + pLacz : p ⁇ 0.05 logrank MCP-4 opp: hMCP-4 injected at distant site.]
  • Examples 7-9 thus indicate that the chemokine hMCP-4 can be used as adjuvant of immune responses, in particular anti-tumor responses.
  • the enhanced immune response as mediated by MCP-4 administration has been measured as enhanced antigen-specific immunoglobulin levels in serum.
  • B cell responses to MCP-4 administration.
  • immunoglobulins such as lgG2a, that require T Cell mediated help for switch, it is likely that there is an increase in T Cell mediated responses as well.
  • human 6Ckine (h ⁇ Ckine) is a chemotactic factor for all known subsets of dendritic cells in man, in vitro.
  • h6Ckine is active on human blood dendritic cells following a short 3 hour incubation with GM-CSF, IL-3 and CD40L. (Fig. 8)
  • CD34-derived DC precursors were isolated by Facs-sorting according to CD1a and CD14 expression after 6 days of culture in presence of GM-CSF+TNF and SCF. Cells were cultured until day 12 in GM-CSF alone (immature) or GM-CSF+CD40-L (mature) for the last two days.
  • Monocyte-derived DC were generated by culturing monocytes in presence of GM-CSF+IL-4 for 5 days and activated (mature) or not (immature) with CD40-L for the last 2 days.
  • CD11c+ DC and CD11c- plasmacytoid DC were cultured for three hours in presence of GM-CSF+IL-3 with (mature) or without (immature) CD40- L.
  • Migration assays were carried out during 1 to 3 hours using 5 or 8 ⁇ m pore size Transwell (6.5mm diameter, COSTAR, Cambridge, MA), and revealed by facs analysis. All populations respond to 6Ckine but only after CD40-L activation.
  • C26 colon carcinoma tumor cells engineered to express m6Ckine are less tumorigenic and that this effect depends on CD8+ cells and Natural Killer cell activity, in vivo. (Fig. 9);
  • mice 6- to 10-week-old female BALB/c (H-2 d ) mice were purchased from Charles
  • Fig. 10 shows that subcutaneous C26-6CK cell injection results in significantly delayed tumor intake compared to parental tumor cells (p ⁇ 0.01 ) by logrank analysis (A and B: C26 + control vs C26-6CK + control).
  • CD8+ cells A or Natural Killer cell activity (B) with specific antibodies in vivo partially reverts the delayed tumorigenicity of the C26-6CK tumor cells, indicating that CD8+ cells and NK cells play a role in delaying tumor growth.
  • Tumors were surgically removed when reaching an approximate size of 1 cm.
  • the tumor mass was minced into small fragments and incubated in collagenase A (Roche Molecular Biochemicals) solution for 30 min at 37°C under agitation.
  • the suspension was then washed several times in DMEM. Staining of cell suspensions was performed in PBS + 5% FCS.
  • Fc receptors Prior to incubation with FITC-, biotin- or PE- labeled specific antibodies, Fc receptors were blocked using Fc-BlockTM CD16/CD32 antibody (PharMingen, San Diego, CA).
  • the various antibodies all from
  • PharMingen used in this study were CD8 ⁇ (53-5.8), CD11c (HL3), anti-MHC class II l-A d /l-E d (269), CD3 (145-2C11). Biotinylated antibodies were revealed with PE- streptavidin (Becton Dickinson). Phenotypic parameters were acquired on a FacScan (Becton Dickinson, Mountain View, CA) and analyzed using the CellQuest software (Becton Dickinson). In Fig.
  • Data show a significant recruitment of both leukocyte subsets in C26-6CK tumors compared to C26 tumors (Student's t test).
  • tumors were removed from animals and embedded in OCT compound (Miles laboratory, Elkhart, IN) before being snap frozen in liquid nitrogen and store at -80°C until immunohistochemistry procedures.
  • OCT compound Miles laboratory, Elkhart, IN
  • Five- micrometer cryostat sections applied onto glass slides were fixed in acetone and incubated with 1% H 2 0 2 for 10 min at room temperature. Slides were then incubated with the biotin-blockTM and avidin-blockTM reagents (both from Vector, Burlingame, CA). All incubations were followed by three 2 min-washes in PBS (Gibco-BRL). Slides were then pre-incubated for 30 min with a 1/10 dilution of serum from the same species of the secondary antibody (Dako, Glostrup, Denmark). Slides were then incubated sequentially with 5 ⁇ g/ml of purified CD105 (clone MJ7/18,
  • chemokine 6Ckine could be used in cancer treatment through gene transfer.
  • Preferred embodiments consist of but are not restricted to: DNA or viral vector (e.g. adenovirus) encoding for m6Ckine or h ⁇ Ckine or fraction of m ⁇ Ckine or h6Ckine, with or without a targeting moiety (peptide or antibody).
  • mice 6- to 10-week-old female BALB/c (H-2 d ) mice were purchased from Charles
  • mice were injected intra-tumorally with 10 ng >97% pure recombinant human or mouse 6Ckine/SLC (R&D Systems, Minneapolis, MN) under 50 ⁇ l PBS.
  • Figure 1 shows that mice injected with h ⁇ Ckine or m ⁇ Ckine show improvement in survival compared with PBS vehicle alone (A). Injection of h6Ckine also decreased the growth of tumors (B).
  • mice Female mice (BALB/c ByJ; Jackson Laboratories) were injected by subcutaneous route with 3x10 15 4T1-p53 mammary tumor cells (syngeneic) in a volume of 0.2 ml (medium) into the left flank of animals. Animals received an intratumoral injection when the tumor grew to a size of 50-100 mm 3 of 100 ⁇ l of CMCB (1e10 PN/injection) in VPBS. Mice received 3 injections per week (Monday, Wednesday, Friday) for two weeks. The tumors were measured three times weekly using a caliper (length, width, depth), the tumor volume was calculated according to formula:
  • mice from each group were sacrificed, starting at the time when the tumors reached 50mm 3 (typically day 10), and the tumors and lungs were resected for tissue processing for the biochemical analyses described below and to assess the presence of metastases by gross and histological means.
  • 6Ckine inhibits tumor growth and spontaneous metastasis by in established tumors by augmenting immunity and suppressing angiogenesis.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Oncology (AREA)
  • Epidemiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Communicable Diseases (AREA)
  • Developmental Biology & Embryology (AREA)
  • Reproductive Health (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Urology & Nephrology (AREA)
  • Cell Biology (AREA)
  • Pregnancy & Childbirth (AREA)
  • Gynecology & Obstetrics (AREA)
  • Pulmonology (AREA)
  • Molecular Biology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Ophthalmology & Optometry (AREA)
  • Endocrinology (AREA)
  • Neurosurgery (AREA)
  • Neurology (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Dermatology (AREA)

Abstract

Selon la présente invention, les cellules dendritiques jouent un rôle déterminant dans les réponses immunitaires spécifiques aux antigènes. Cette invention concerne des substances et des méthodes destinées à traiter des états pathologiques, tels que le cancer et les maladies auto-immunes, en favorisant ou en inhibant la migration ou l'activation de cellules dendritiques de présentation antigénique. Plus particulièrement, on utilise des chimiokines pour induire, amplifier ou moduler une réponse immunitaire. Dans un mode de réalisation, ces chimiokines sont utilisées pour attirer des cellules dendritiques vers le site d'administration d'antigènes. Un nombre accru de cellules dendritiques au niveau du site d'administration d'antigènes indique une augmentation de l'absorption antigénique et une modification de la réponse immunitaire.
PCT/US2002/001849 2001-01-24 2002-01-22 Chimiokines utilisees comme adjuvants de la reponse immunitaire WO2002058723A2 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
EP02705895A EP1392351A2 (fr) 2001-01-24 2002-01-22 Chimiokines utilisees comme adjuvants de la reponse immunitaire
MXPA03006587A MXPA03006587A (es) 2001-01-24 2002-01-22 Quimiocinas como adyuvantes de respuesta inmune.
CA002434320A CA2434320A1 (fr) 2001-01-24 2002-01-22 Chimiokines utilisees comme adjuvants de la reponse immunitaire
JP2002559057A JP2004520043A (ja) 2001-01-24 2002-01-22 免疫応答のアジュバントとしてのケモカイン

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US09/768,917 2001-01-24
US09/768,917 US7217700B2 (en) 1998-07-16 2001-01-24 Chemokines as adjuvants of immune response

Publications (2)

Publication Number Publication Date
WO2002058723A2 true WO2002058723A2 (fr) 2002-08-01
WO2002058723A3 WO2002058723A3 (fr) 2003-12-18

Family

ID=25083870

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2002/001849 WO2002058723A2 (fr) 2001-01-24 2002-01-22 Chimiokines utilisees comme adjuvants de la reponse immunitaire

Country Status (5)

Country Link
EP (1) EP1392351A2 (fr)
JP (2) JP2004520043A (fr)
CA (1) CA2434320A1 (fr)
MX (1) MXPA03006587A (fr)
WO (1) WO2002058723A2 (fr)

Cited By (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003035106A1 (fr) * 2001-10-24 2003-05-01 Chemocentryx Procedes et compositions servant a induire une reponse immune
WO2005108584A1 (fr) * 2004-05-10 2005-11-17 Dongguan Haofa Bio-Engineering Development Co., Ltd. Vaccin chimiokine-antigene recombinant
JP2006517970A (ja) * 2003-02-14 2006-08-03 ユニバーシティ オブ サザン カリフォルニア 癌免疫療法のための組成物及び方法
JP2007525444A (ja) * 2003-03-25 2007-09-06 ヘリコニア コーポレイション 治療応答を誘導するための組成物
US9370558B2 (en) 2008-02-13 2016-06-21 President And Fellows Of Harvard College Controlled delivery of TLR agonists in structural polymeric devices
US9381235B2 (en) 2009-07-31 2016-07-05 President And Fellows Of Harvard College Programming of cells for tolerogenic therapies
US9446107B2 (en) 2005-12-13 2016-09-20 President And Fellows Of Harvard College Scaffolds for cell transplantation
US9486512B2 (en) 2011-06-03 2016-11-08 President And Fellows Of Harvard College In situ antigen-generating cancer vaccine
US9675561B2 (en) 2011-04-28 2017-06-13 President And Fellows Of Harvard College Injectable cryogel vaccine devices and methods of use thereof
US9770535B2 (en) 2007-06-21 2017-09-26 President And Fellows Of Harvard College Scaffolds for cell collection or elimination
WO2017190074A1 (fr) * 2016-04-28 2017-11-02 The University Of Chicago Lymphangiogenèse pour l'immunomodulation thérapeutique
US9821045B2 (en) 2008-02-13 2017-11-21 President And Fellows Of Harvard College Controlled delivery of TLR3 agonists in structural polymeric devices
US9937249B2 (en) 2012-04-16 2018-04-10 President And Fellows Of Harvard College Mesoporous silica compositions for modulating immune responses
US10045947B2 (en) 2011-04-28 2018-08-14 President And Fellows Of Harvard College Injectable preformed macroscopic 3-dimensional scaffolds for minimally invasive administration
US10682400B2 (en) 2014-04-30 2020-06-16 President And Fellows Of Harvard College Combination vaccine devices and methods of killing cancer cells
US11150242B2 (en) 2015-04-10 2021-10-19 President And Fellows Of Harvard College Immune cell trapping devices and methods for making and using the same
US11202759B2 (en) 2010-10-06 2021-12-21 President And Fellows Of Harvard College Injectable, pore-forming hydrogels for materials-based cell therapies
US11419928B2 (en) 2017-10-25 2022-08-23 The Johns Hopkins University Methods and compositions for treating cancer
US11555177B2 (en) 2016-07-13 2023-01-17 President And Fellows Of Harvard College Antigen-presenting cell-mimetic scaffolds and methods for making and using the same
US11732029B2 (en) 2017-11-13 2023-08-22 The University Chicago Methods and compositions for the treatment of wounds
US11752238B2 (en) 2016-02-06 2023-09-12 President And Fellows Of Harvard College Recapitulating the hematopoietic niche to reconstitute immunity
US11786457B2 (en) 2015-01-30 2023-10-17 President And Fellows Of Harvard College Peritumoral and intratumoral materials for cancer therapy

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030077247A1 (en) * 2001-09-20 2003-04-24 Schering Corporation Chemokines as adjuvants of immune response
CA2448995A1 (fr) * 2003-11-12 2005-05-12 James Keenan Dispositif et methode pour attirer les cellules malades et les substances etrangeres

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999046392A1 (fr) * 1998-03-12 1999-09-16 The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Methodes et compositions utilisant des proteines de fusion chemokine-antigene tumoral comme vaccins anti-cancer
EP0974357A1 (fr) * 1998-07-16 2000-01-26 Schering-Plough Chemokines comme adjuvants de la réponse immunitaire

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999046392A1 (fr) * 1998-03-12 1999-09-16 The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Methodes et compositions utilisant des proteines de fusion chemokine-antigene tumoral comme vaccins anti-cancer
EP0974357A1 (fr) * 1998-07-16 2000-01-26 Schering-Plough Chemokines comme adjuvants de la réponse immunitaire

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
CAUX C ET AL: "DENDRITIC CELL BIOLOGY AND REGULATION OF DENDRITIC CELL TRAFFICKINGBY CHEMOKINES" SPRINGER SEMINAR IN IMMUNOPATHOLOGY, SPRINGER VERLAG, DE, vol. 22, no. 4, 2000, pages 345-369, XP000982553 ISSN: 0344-4325 *
DIEU-NOSJEAN MARIE-CAROLINE ET AL: "Regulation of dendritic cell trafficking: A process that involves the participation of selective chemokines." JOURNAL OF LEUKOCYTE BIOLOGY, vol. 66, no. 2, August 1999 (1999-08), pages 252-262, XP008012618 ISSN: 0741-5400 *
KELLERMANN S A ET AL: "The CC chemokine receptor-7 ligands 6Ckine and macrophage inflammatory protein-3 beta are potent chemoattractants for in vitro- and in vivo-derived dendritic cells." JOURNAL OF IMMUNOLOGY (BALTIMORE, MD.: 1950) UNITED STATES 1 APR 1999, vol. 162, no. 7, 1 April 1999 (1999-04-01), pages 3859-3864, XP002227532 ISSN: 0022-1767 *
NAKAMURA KOICHIRO ET AL: "Keratinocyte-derived monocyte chemoattractant protein 1 (MCP-1): Analysis in a transgenic model demonstrates MCP-1 can recruit dendritic and Langerhans cells to skin." JOURNAL OF INVESTIGATIVE DERMATOLOGY, vol. 105, no. 5, 1995, pages 635-643, XP008009489 ISSN: 0022-202X *
VECCHI A ET AL: "DIFFERENTIAL RESPONSIVENESS TO CONSTITUTIVE VS. INDUCIBLE CHEMOKINES OF IMMATURE AND MATURE MOUSE DENDRITIC CELLS" JOURNAL OF LEUKOCYTE BIOLOGY, FEDERATION OF AMERICAN SOCIETIES FOR EXPERIMENTAL, US, vol. 66, no. 3, 1999, pages 489-494, XP000906790 ISSN: 0741-5400 *
XU LOU LING ET AL: "Human recombinant monocyte chemotactic protein and other c-c chemokines bind and induce directional migration of dendritic cells in vitro." JOURNAL OF LEUKOCYTE BIOLOGY, vol. 60, no. 3, 1996, pages 365-371, XP008009488 ISSN: 0741-5400 *
ZHU KEJIAN ET AL: "Human monocyte-derived dendritic cells expressing both chemotactic cytokines IL-8, MCP-1, RANTES and their receptors, and their selective migration to these chemokines." CHINESE MEDICAL JOURNAL (ENGLISH EDITION), vol. 113, no. 12, December 2000 (2000-12), pages 1124-1128, XP008009490 ISSN: 0366-6999 *

Cited By (39)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003035106A1 (fr) * 2001-10-24 2003-05-01 Chemocentryx Procedes et compositions servant a induire une reponse immune
JP2006517970A (ja) * 2003-02-14 2006-08-03 ユニバーシティ オブ サザン カリフォルニア 癌免疫療法のための組成物及び方法
JP2007525444A (ja) * 2003-03-25 2007-09-06 ヘリコニア コーポレイション 治療応答を誘導するための組成物
AU2005240691B2 (en) * 2004-05-10 2009-06-04 Cancer Institute, Chinese Academy Of Medical Sciences Recombinant chemokine-antigen vaccine
EP1748075A1 (fr) * 2004-05-10 2007-01-31 Dongguan Bio-Engineering Development Co. Ltd. Vaccin chimiokine-antigene recombinant
EP1748075A4 (fr) * 2004-05-10 2008-01-02 Cancer Inst Chinese Academy Of Vaccin chimiokine-antigene recombinant
JP4838238B2 (ja) * 2004-05-10 2011-12-14 キャンサー インスティテュート，チャイニーズ アカデミー オブ メディカル サイエンシーズ 組換え走化抗原遺伝子ワクチン
US8133873B2 (en) 2004-05-10 2012-03-13 Cancer Institute, Chinese Academy Of Medical Sciences Recombinant chemokine-antigen vaccine
WO2005108584A1 (fr) * 2004-05-10 2005-11-17 Dongguan Haofa Bio-Engineering Development Co., Ltd. Vaccin chimiokine-antigene recombinant
US11096997B2 (en) 2005-12-13 2021-08-24 President And Fellows Of Harvard College Scaffolds for cell transplantation
US10149897B2 (en) 2005-12-13 2018-12-11 President And Fellows Of Harvard College Scaffolds for cell transplantation
US10137184B2 (en) 2005-12-13 2018-11-27 President And Fellows Of Harvard College Scaffolds for cell transplantation
US9446107B2 (en) 2005-12-13 2016-09-20 President And Fellows Of Harvard College Scaffolds for cell transplantation
US10695468B2 (en) 2007-06-21 2020-06-30 President And Fellows Of Harvard College Scaffolds for cell collection or elimination
US9770535B2 (en) 2007-06-21 2017-09-26 President And Fellows Of Harvard College Scaffolds for cell collection or elimination
US10258677B2 (en) 2008-02-13 2019-04-16 President And Fellows Of Harvard College Continuous cell programming devices
US9821045B2 (en) 2008-02-13 2017-11-21 President And Fellows Of Harvard College Controlled delivery of TLR3 agonists in structural polymeric devices
US10568949B2 (en) 2008-02-13 2020-02-25 President And Fellows Of Harvard College Method of eliciting an anti-tumor immune response with controlled delivery of TLR agonists in porous polymerlc devices
US9370558B2 (en) 2008-02-13 2016-06-21 President And Fellows Of Harvard College Controlled delivery of TLR agonists in structural polymeric devices
US10328133B2 (en) 2008-02-13 2019-06-25 President And Fellows Of Harvard College Continuous cell programming devices
US10080789B2 (en) 2009-07-31 2018-09-25 President And Fellows Of Harvard College Programming of cells for tolerogenic therapies
US9381235B2 (en) 2009-07-31 2016-07-05 President And Fellows Of Harvard College Programming of cells for tolerogenic therapies
US11202759B2 (en) 2010-10-06 2021-12-21 President And Fellows Of Harvard College Injectable, pore-forming hydrogels for materials-based cell therapies
US10045947B2 (en) 2011-04-28 2018-08-14 President And Fellows Of Harvard College Injectable preformed macroscopic 3-dimensional scaffolds for minimally invasive administration
US9675561B2 (en) 2011-04-28 2017-06-13 President And Fellows Of Harvard College Injectable cryogel vaccine devices and methods of use thereof
US10406216B2 (en) 2011-06-03 2019-09-10 President And Fellows Of Harvard College In situ antigen-generating cancer vaccine
US9486512B2 (en) 2011-06-03 2016-11-08 President And Fellows Of Harvard College In situ antigen-generating cancer vaccine
US11278604B2 (en) 2012-04-16 2022-03-22 President And Fellows Of Harvard College Mesoporous silica compositions comprising inflammatory cytokines comprising inflammatory cytokines for modulating immune responses
US9937249B2 (en) 2012-04-16 2018-04-10 President And Fellows Of Harvard College Mesoporous silica compositions for modulating immune responses
US10682400B2 (en) 2014-04-30 2020-06-16 President And Fellows Of Harvard College Combination vaccine devices and methods of killing cancer cells
US11786457B2 (en) 2015-01-30 2023-10-17 President And Fellows Of Harvard College Peritumoral and intratumoral materials for cancer therapy
US11150242B2 (en) 2015-04-10 2021-10-19 President And Fellows Of Harvard College Immune cell trapping devices and methods for making and using the same
US11752238B2 (en) 2016-02-06 2023-09-12 President And Fellows Of Harvard College Recapitulating the hematopoietic niche to reconstitute immunity
WO2017190074A1 (fr) * 2016-04-28 2017-11-02 The University Of Chicago Lymphangiogenèse pour l'immunomodulation thérapeutique
US10980877B2 (en) * 2016-04-28 2021-04-20 The University Of Chicago Method for treating melanoma using lymphangiogenesis inducers and a melanoma-specific antigen
US20190099485A1 (en) * 2016-04-28 2019-04-04 The University Of Chicago Lymphangiogenesis for therapeutic immunomodulation
US11555177B2 (en) 2016-07-13 2023-01-17 President And Fellows Of Harvard College Antigen-presenting cell-mimetic scaffolds and methods for making and using the same
US11419928B2 (en) 2017-10-25 2022-08-23 The Johns Hopkins University Methods and compositions for treating cancer
US11732029B2 (en) 2017-11-13 2023-08-22 The University Chicago Methods and compositions for the treatment of wounds

Also Published As

Publication number Publication date
EP1392351A2 (fr) 2004-03-03
WO2002058723A3 (fr) 2003-12-18
MXPA03006587A (es) 2003-09-22
JP2008133296A (ja) 2008-06-12
JP2004520043A (ja) 2004-07-08
CA2434320A1 (fr) 2002-08-01

Similar Documents

Publication Publication Date Title
US20070128159A1 (en) Chemokines as adjuvants of immune response
WO2002058723A2 (fr) Chimiokines utilisees comme adjuvants de la reponse immunitaire
Furumoto et al. Induction of potent antitumor immunity by in situ targeting of intratumoral DCs
Mocellin et al. Cytokines and immune response in the tumor microenvironment
US20070166280A1 (en) Chemokines as adjuvants of immune response
Kirk et al. T cell-dependent antitumor immunity mediated by secondary lymphoid tissue chemokine: augmentation of dendritic cell-based immunotherapy
Yang et al. Intratumoral administration of dendritic cells overexpressing CCL21 generates systemic antitumor responses and confers tumor immunity
Guiducci et al. Redirecting in vivo elicited tumor infiltrating macrophages and dendritic cells towards tumor rejection
Benchetrit et al. Interleukin-17 inhibits tumor cell growth by means of a T-cell–dependent mechanism
Vicari et al. Antitumor effects of the mouse chemokine 6Ckine/SLC through angiostatic and immunological mechanisms
EP1255554B1 (fr) Vaccin contre les tumeurs specifiques du rein, dirige contre l'antigene g-250 de la tumeur du rein
Kim et al. AIMP1/p43 protein induces the maturation of bone marrow-derived dendritic cells with T helper type 1-polarizing ability
WO2002032378A2 (fr) Cellules de fusion et compositions cytokiniques pour le traitement de maladies
AU2002225990A1 (en) Fusion cells and cytokine compositions for treatment of disease
JP2007523901A (ja) 癌治療のための融合細胞とインターロイキン−１２の複合免疫療法
Hüttner et al. Generation of potent anti-tumor immunity in mice by interleukin-12-secreting dendritic cells
US20030008840A1 (en) Methods for treating cancer
Liu et al. Dendritic cells engineered to express the Flt3 ligand stimulate type I immune response, and induce enhanced cytoxic T and natural killer cell cytotoxicities and antitumor immunity
Wang et al. Potential involvement of monocyte chemoattractant protein (MCP)-1/CCL2 in IL-4-mediated tumor immunity through inducing dendritic cell migration into the draining lymph nodes
US20020114812A1 (en) Methods and compositions for modulating regulation of the cytotoxic lymphocyte response by macrophage migration inhibitory factor
Pajtasz-Piasecka et al. Generation of anti-tumor response by JAWS II mouse dendritic cells transduced with murine interleukin 12 genes
Kwon et al. Co-expression of bfl-1 enhances host response in the herpes simplex virus-thymidine kinase/ganciclovir gene therapy system
Neeson et al. Effects of the tumor microenvironment on the efficacy of tumor immunotherapy
WO2023192542A1 (fr) Compositions et procédés pour activation de cellules immunitaires
Puskas et al. Chemokine Gene-Modified Dendritic Cells for Cancer Therapy

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CZ DE DK DM DZ EC EE ES FI GB GD GE HR HU ID IL IN IS JP KG KR KZ LC LK LR LT LU LV MA MD MG MK MN MX MZ NO NZ PH PL PT RO RU SE SG SI SK SL TJ TM TN TR TT TZ UA UZ VN YU ZA ZM

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 2002705895

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2434320

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: PA/a/2003/006587

Country of ref document: MX

WWE Wipo information: entry into national phase

Ref document number: 2002559057

Country of ref document: JP

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

WWP Wipo information: published in national office

Ref document number: 2002705895

Country of ref document: EP