WO2005037866A2 - Igg3 anti rhesus-d dans la lignee yb2/0 ayant une forte activite en phagocytose - Google Patents
Igg3 anti rhesus-d dans la lignee yb2/0 ayant une forte activite en phagocytose Download PDFInfo
- Publication number
- WO2005037866A2 WO2005037866A2 PCT/FR2004/002657 FR2004002657W WO2005037866A2 WO 2005037866 A2 WO2005037866 A2 WO 2005037866A2 FR 2004002657 W FR2004002657 W FR 2004002657W WO 2005037866 A2 WO2005037866 A2 WO 2005037866A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- igg3
- use according
- iggl
- produced
- antibodies
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/34—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against blood group antigens
-
- 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
- A61P31/04—Antibacterial agents
- A61P31/06—Antibacterial agents for tuberculosis
-
- 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
- A61P31/08—Antibacterial agents for leprosy
-
- 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
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- 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
- A61P31/14—Antivirals for RNA viruses
- A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
-
- 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
- A61P31/14—Antivirals for RNA viruses
- A61P31/18—Antivirals for RNA viruses for HIV
-
- 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
- A61P31/20—Antivirals for DNA viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P33/00—Antiparasitic agents
- A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
-
- 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
- A61P35/02—Antineoplastic agents specific for leukemia
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
-
- 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
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/40—Immunoglobulins specific features characterized by post-translational modification
- C07K2317/41—Glycosylation, sialylation, or fucosylation
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Definitions
- the present invention relates to the use of chimeric, humanized or human IgG3 class monoclonal antibodies produced in a rat myeloma cell line, in particular YB2 / 0 (ATCC n ° CRL 1662) or a line derived or modified from YB2 / 0, for the preparation of a medicament intended for the treatment of various cancerous and infectious pathologies.
- YB2 / 0 ATCC n ° CRL 1662
- a line derived or modified from YB2 / 0 for the preparation of a medicament intended for the treatment of various cancerous and infectious pathologies.
- These antibodies exhibit strong phagocytosis activity and can be administered to treat cancers and infections.
- IgG3 notably have specific effector capacities and certainly play an important role in vivo. Although representing only 7% of IgG in human plasma, their proportion is increased during certain immune responses, for example after certain viral infections (Basic and clinical aspects of IgG subclasses. Volume editor, F. Shakib. Basel; New York: Karger, 1986 (Monographs in AUlergy; Vol 19. Pages 122-133), parasites (J Infect Dis. 2003 Mar l; 187 (5): 862-5, 2003. Immunoglobulin G (IgG) responses to Plasmodium falciparum glycosylphosphatidylinositols are short-lived and predominantly of the IgG3 subclass.
- the YB2 / 0 line has been selected for several years for its capacity to confer on the IgG1s produced improved functional properties.
- CD 16 Fc III receptor
- the glycan structures of said antibodies are of the biantennary type, characterized by short chains, low sialylation and low fucosylation.
- CD16 has the advantage of also inducing the production of cytokines, in particular the production of IFN and / or other cytokines or chemokines.
- the two characteristics mentioned above complement each other. Indeed, the production of IFN ⁇ or other cytokines and / or chemokines by effector cells induced by the selected antibodies can reinforce the therapeutic effect by stimulating effector mechanisms of the immune system other than ADCC in the treated patients. The mechanism of action of such stimulation is probably due to an autocrine upregulation of the effector cells. It can be postulated that the antibodies binding to CD 16 induce cytotoxic activity as well as the production of IFN ⁇ or other cytokines / chemokines which ultimately leads to an even greater increase in cytotoxic activity.
- anti-D IgG3 has been expressed in a rat myeloma line in order to assess whether this line, in particular YB2 / 0, can confer on the antibodies produced improved functional properties, as is the case for IgGl.
- the invention relates to chimeric, humanized or human monoclonal antibodies of class IgG3 characterized in that they are produced in a rat myeloma cell line.
- said IgG3 are produced in the line YB2 / 0 (ATCC n ° CRL 1662) or a line derived or modified from YB2 / 0.
- the glycan structure of the Fc region corresponds to a biantennary type, with short chains, weak sialylation and weak fucosylation.
- the level of intermediate GlcNac is not zero.
- Such antibodies are more particularly selected from the forms:
- the invention relates to monoclonal antibodies of class IgG3 in which the fucose content is less than 65%, 60%, 50%, 40%, or 35%.
- the fucose content is between 20% and 45% or even between 25% and 40%.
- the fucose content is less than 35%.
- the invention also relates to antibodies of class IgG3 having the glycosylation profile indicated above produced in equivalent biological systems, in particular in genetically modified or transformed non-human cells, plants or animals, for example by introduction of a sequence expressing a or many glycosyl transferase (s) so as to obtain antibodies having a profile essentially similar to the glycosylation profile obtained in YB 2 / O.
- s glycosyl transferase
- the IgG3 produced in the YB2 / 0 line have, relative to those produced in lines like CHO for example, specific functional characteristics: a) a strong binding to CD 16 which is comparable to that of the IgG1 produced in the same cell line. b) an ability to induce an inhibition of cytokine secretion induced by IgGl. c) a greater capacity of the IgG3 produced in YB2 / 0 to induce the secretion of gamma IFN, IL6 and TNF alpha than the IgG3 produced in CHO; and a lower capacity of IgG3 YB2 / 0 to induce the secretion of TNF alpha and IFN gamma than IgG1 YB2 / 0. d) potentiation of phagocytosis.
- the antibody of class IgG3 of the invention can be selected by way of example from the antibodies directed against: CD2, CD3, CD4, CD5, CD7, CD8, CDU, CD18, CD19, CD20, CD25, CD45 and CD52 as Campath-IH®, CD30, CD33, CD38 or CD44.
- Other antibodies can be selected from anti Ep-CAM, anti HER2, anti HER1, anti GD3, anti CA125, anti GD, anti GD2, anti CD-23 and anti ProteinC; anti-KIR3DL2, anti-EGFR, specific anti-idiotypes of inhibitors for example of coagulation factors, anti-virals: H1N, HBN, HCN and RSN.
- the invention relates to a process for the production of chimeric, humanized or human monoclonal antibodies of class IgG3 having the functional characteristics mentioned above comprising the transfection of a cell line of rat myeloma preferably, the line YB2 / 0 (ATCC No. CRL 1662) or a line derived or modified from YB2 / 0, with one or more vector (s) comprising the coding sequences for the heavy and light chains of antibodies to class IgG3, the expression of said antibodies in the transfected cell line, the extraction and purification of the antibodies.
- a cell line of rat myeloma preferably, the line YB2 / 0 (ATCC No. CRL 1662) or a line derived or modified from YB2 / 0, with one or more vector (s) comprising the coding sequences for the heavy and light chains of antibodies to class IgG3, the expression of said antibodies in the transfected cell line, the extraction and purification of the antibodies.
- a system with two expression vectors for example vectors derived from RSN
- one coding for the heavy chains and the other coding for the light chains is used, one coding for the heavy chains and the other coding for the light chains.
- a different selection marker is present in each vector. Specific constructions are shown in Figure 1.
- the invention also relates to the system described above in which the heavy and light chains are produced in equimolar amount.
- the construction of expression vectors can be implemented according to procedures known to those skilled in the art (Molecular Cloning: A Laboratory Manual, Second
- Co-transfection into the rat myeloma line of the two vectors can be carried out by means of equimolar amount by standard procedures of the calcium phosphate or lipofectin precipitation type. Then, the transfected lines are selected in the appropriate culture media.
- the invention relates to rat myeloma cell lines, in particular YB2 / 0 and derived lines, transfected with one or more vector (s) allowing the expression of a functional IgG3.
- the invention also relates to cells having been transfected with one or more vector (s) described above. These cells are characterized in that they produce IgG3 having the glycosylation profile mentioned above and at least one of the properties a) to d) described above.
- a cell derived from a line described above is also an object of the invention.
- the invention relates to the use of IgG3 described above, in particular of IgG3 expressed in YB2 / 0, for the preparation of a medicament.
- the invention relates to the use of chimeric, humanized or human monoclonal antibodies of class IgG3, produced in a rat myeloma cell line, in particular YB2 / 0 (ATCC N ° CRL 1662) or a line derived or modified of YB2 / 0, for the preparation of a medicament intended for the treatment of various cancerous pathologies, or of various infectious pathologies of infectious viral, bacterial or parasitic origin.
- these antibodies can be used for the preparation of a medicament intended for the prevention of fetal-maternal immunization.
- the patients concerned are patients who are weak responders to treatment with an IgG1 or an IgG3 expressed in CHO.
- “Weak responder patients” is understood to mean treated patients with a so-called stable condition, with less than 50% reduction and less than 25% increase in lesions, and no new lesions. This group of patients also includes patients for whom no response is observed (progression of the disease which can lead to death). For infectious diseases, these patients correspond to patients for whom the viral or bacterial load is reduced by less than 50% by conventional treatment.
- the antibody can be used in late diagnosed patients.
- the cancer pathologies which can be treated in a particularly advantageous manner by means of the antibodies according to the invention are chosen from the group comprising neuroectodermal tumors, colorectal cancers, melanomas, breast cancer, leukemias, in particular HCL (Hairy Cell LeuOkemia), lymphomas such as DLBCL (Primary Diffuse Large B-Cell Lympriomas), acute leukemias, osteosarcomas, cancer, in particular poximon cancer, this list is not exhaustive.
- the cancer pathologies treated according to the invention are associated with infections of viral or bacterial origin, such as prostate cancer (Lightfoot N, Conlon M, Kreiger N, Sass-Kortsak A, Purdham J, Darlington G. Medical history, sexual, and maturational factors and prostate cancer risk. Ann Epidemiol. 2004 Oct; 14 (9): 655-662; Huycke MM, Gaskins HR. Commensal bacteria, redox stress, and colorectal cancer: mechanisms and models. Exp Biol Med (Maywood). 2004 Jul; 229 (7): 586-97), leukemia, or E aposi's sarcoma.
- infectious agents found in infectious diseases associated with cancer can be Candida, Achromobacter or Alcaligenes (Aisenberg G, Rolston KN, Safdar A. Bacteremia caused by Achromobacter and Alcaligenes species in 46 patients with cancer (1989-2003). Cancer. 2004 Sep 23; Boktour MR, Kontoyiannis DP, Hanna HA, Hachem RY, Girgawy E, Bodey GP, Raad IL Multiple-species candidemia in patients with cancer. 2004 Aug 31;) or the Epstein Barr virus.
- infectious pathologies advantageously treated with the antibody according to the invention, mention may be made of diphtheria, viral hemorrhagic fevers, typhoid fever, influenza, hepatitis B and C, respiratory infections due to RSN, NIH infections and at CMN, Legionellosis, Leishmaniasis, leprosy, rabies, AIDS or even tuberculosis, this list is not exhaustive.
- the IgG3s of the invention are of interest with regard to these uses because of their strong attachment to the low affinity Fc receptor (CD 16) and / or their capacity to induce phagocytosis.
- the medicament according to the invention is intended to be used in combination with an IgG1.
- the use according to the invention of IgG3 as described above is particularly advantageous in this aspect of the invention for the capacity of these IgG3 to negatively modulate the secretion of cytokines induced by IgGl, in particular the levels of IFN gamma, TNF alpha and / or IL6.
- the IgG3s as described above are used for the preparation of a medicament for the treatment of cancer pathologies in patients having a "cytokine release syndrome", in particular in patients treated with a IgGl produced in YB2 / 0.
- This application takes advantage of the ability of said IgG3s to negatively modulate the secretion of cytokines.
- hypothermia acute renal necrosis and liver diseases due to the “cytokine release syndrome” induced by the administration of an anti-CD3 monoclonal antibody, for example 145-2C11 ( Alegre ML et al, J Immunol. 1991 Feb 15; 146 (4): 1184-91; Chatenoud L.
- Anti-CD3 antibodies towards clinical antigen-specific immunomodulation. Curr Opin Pharmacol. 2004 Aug; 4 (4): 403- 7; Yamada-Ohnishi Y, Azuma H, Urushibara N, Yamaguchi M, Fujihara M, Kobata T, Ikeda H. Cytotoxic Difference of T Cells Expanded with Anti-CD3 Monoclonal Antibody in the Présence and Absence of Anti- CD28 Monoclonal Antibody. Stem Cells Dev. 2004 Jun; 13 (3): 315-22.).
- the invention relates to the use of an IgG3 described above, which can be an anti-CD20, to prevent the appearance of the "cytokine-release syndrome" in patients treated with Rituximab® (IDEC-C2B8 ); Winkler U et al, Cytokine-release syndrome in patients with B-cell chronic lymphocytic leukemia and high lymphocyte counts after treatment with an anti-CD20 monoclonal antibody, Blood. 1999 Oct 1; 94 (7): 2217-24.
- the IgG3 of the invention is useful for preventing the undesirable effects of the CAMPATH® antibody or OKT3.
- CAMPATH 1-H which binds to CD52 on lymphocytes and monocytes, induces the release of TNF, IFN and IL-6 leading to the "cytokine-release syndrome", Mark G. Wing et al, Mechanism of First-Dose Cytokine-Release Syndrome by CAMPATH 1-H: Involvement of CD16 (FcRIII) and CDlla / CD18 (LFA-1) on NK Cells, J. Clin. Invest, Volume 98, Number 12, December 1996, 2819-2826.
- OKT3 which binds to CD3, has also been described as inducing cytokine secretion syndrome (First MR, Schroeder TJ, Hariharan S. OKLT3- induced cytokine-release syndrome: renal effects (cytokine nephropathy). Transplant Proc. 1993 Apr; 25 (2 Suppl l): 25-6).
- Another object of the invention is to provide a method for modulating the secretion of cytokines induced by an IgGl by adding to the biological system containing said
- IgGl of IgG3 produced in a rat myeloma cell line, in particular
- the IgG1s whose cytokine secretion is modulated are produced in a rat myeloma cell line, in particular in YB2 / 0.
- the invention relates to a pharmaceutical composition of therapeutic antibodies comprising IgG1, IgG3 and at least one excipient.
- at least one of these IgGs is produced in a cell line of rat myeloma, in particular YB2 / 0.
- FIG. 3 Interaction with Jurkat CD16 cells of the coated antibodies on the red cells fixed in the microtiter plate.
- the x-axis represents the fixation of the antibodies on the red cells and the y-axis represents the fixation on the CD 16.
- FIG. 4 IgG3 binding to Jurkat CD16 cells in the absence of targets.
- Figure 5 IL-2 secretion induced by IgGl and IgG3 expressed in YB2 / 0 after interaction with Jurkat CD16 cells.
- Figure 7 ADCC activity in the presence of NK cells and anti-D antibodies
- Figure 8 Secretion of IL-2 by Jurkat CD16 cells induced by IgGl and
- Anti-Rhesus IgG3 (red cells in suspension). Effect of the addition of the various IgG3s on the secretion of IL21 induced by the IgGl YB2 / 0.
- NK or monocytes Figure 10 Percentage of THP 1 cells having phagocytosed one or more red blood cells
- Example 1 Obtaining various anti-D antibodies of class IgG3.
- the construction of the expression vectors to produce two recombinant antibodies is presented in FIG. 1.
- transformants producing IgG3 D29 with anti-D specificity were obtained in the lines YB2 / 0 and CHO.
- FIG. 2 The different antibodies thus produced are shown diagrammatically in FIG. 2:
- - Antibody 2 IgG3 expressed in the CHO line (reference line for the industrial production of recombinant proteins), D29-CHO.
- - Antibody 3 IgG3 (D29, produced by a B lymphocyte fused with P3x229), D29- P3X229
- Example 2 Study of the binding of the three antibodies of Example 1 on Jurkat CD16 cells (CFC).
- This test was set up to assess the ability of anti-D antibodies to bind to the CD 16 receptor (Fc gamma RHIa) expressed on Jurkat CD 16 cells.
- the first step consists in reacting the anti-D antibody with the Rhesus antigens expressed on the surface of Rhesus positive red cell membranes previously coated on 96-well plates with round bottom (fixation by the Fab). This fixation is simultaneously detected by an anti-human IgG antibody labeled with alkaline phosphatase.
- the second step (after fixing the antibody to its antigen), consists in adding Jurkat CD 16 cells which will be able to interact with the Fc part of the antibodies. After centrifugation, a score (fixation index of 1 to 3) corresponding to the Jurkat CD 16 cells which are fixed on the antibodies is estimated visually.
- an IgG3 in the YB2 / 0 line gives it an ability to bind to CD 16 which is comparable to that of an IgG1 expressed in the same cell (R297 expressed in YB2 / 0) and also comparable to that of an IgG3 purified from anti-D polyclonal antibodies.
- Example 3 Measurement by competition of the binding of the three antibodies of Example 1 to Jurkat CD16 cells by flow cytometry.
- Example 4 Measurement of the activation of Jurkat CD16 cells (experimental continuation of Example 2).
- FIG. 5 show that the interaction of IgG3 with Jurkat CD16 induces a much weaker secretion of IL2 than in the presence of IgGl.
- PIgGl YB2 / 0 opposite to IgG3 YB2 / 0 induces from the first fixation indices a secretion of IL2; however, the dose response curve obtained with IgG3 is less than that obtained with IgG1. Only a strong interaction of IgG3 YB2 / 0 with CD 16 (maximum binding index of 3) induces secretion of IL2 comparable to that obtained with the IgG1 produced by YB2 / 0.
- Example 5 Study of the cytotoxicity induced by anti-D antibodies against Rh positive red cells in the presence of PBMC or purified NK cells.
- PBMC cytolysis test quantifies the ability of antibodies to lyse Rhesus positive red cells in the presence of human mononuclear cells (PBMC) and polyvalent immunoglobulins (Tegeline).
- PBMC human mononuclear cells
- Tegeline polyvalent immunoglobulins
- the cytolytic activity of IgG3 expressed in CHO is comparable to that obtained with the antibody expressed by the fused B lymphocyte D29-P3X229.
- an IgG3 in YB2 / 0 potentiates its capacities to induce a lysis of red blood cells in the presence of mononucleated cells (PBL) by comparison with the same antibody produced in CHO or by heteromyeloma.
- the increase in the cytolytic activity of IgG3 expressed in YB20 is 2.8 times greater and also comparable to that induced by the polyclonal fraction IgG3 of WinRho. Nevertheless, the cytolytic activity of the IgG3 produced in YB2 / 0 is lower than that of the IgG1 produced in YB2 / 0 and of the anti-D polyclonal antibody (Poly-D WinRho).
- IgG3 YB2 / 0 induces lysis of red blood cells (55%) 5.5 times greater than that obtained with the same antibody produced in CHO (10%).
- the antibody produced by heteromyeloma P3X229 gives the lowest value (4%).
- the cytolytic activity of the IgG3 produced in YB2 / 0 is lower than that of the IgG1 produced in YB2 / 0 and of the polyclonal antibody WinRho.
- Example 6 Measurement of IL-2 secretion by Jurkat CD16 after fixing of the antibodies on red blood cells in suspension.
- IL2 secretion Jurkat CD 16 cells are incubated with Rhesus positive red cells, IgGl YB2 / 0 or the antibody IgG3 D29 expressed in the various expression systems (YB2 / 0, CHO, B-P3 X229). The secretion of I12 is measured in the supernatants after an overnight incubation by ELISA technique.
- the IgG3 expressed in YB2 / 0 and CHO do not induce secretion of IL2 unlike the IgG1 YB2 / 0, that at identical antibody concentration (FIG. 8).
- the fixation on CD 16 of an IgG3 expressed in YB2 / 0 does not induce IL2 secretion in the presence of red cells in solution and Jurkat CD16 cells .
- Example 4 in which the red cells were coated in microplates, showed that only a strong interaction with CD16 made it possible to induce secretion of IL2.
- the use of more physiological conditions in this example confirm the very low potential of IgG3 YB2 / 0 to induce secretion of IL2 from Jurkat CD 16, unlike IgGl YB2 / 0.
- IgG3 produced in CHO and P3X229 have no effect on the secretion of IL2 induced by IgGl YB2 / 0.
- PIgG3 produced in YB2 / 0 induces a reduction in the induction of IL2 ( Figure 8).
- Example 7 Induction of the secretion of cytokines by anti-D antibodies.
- NK cells or monocytes are incubated with Rhesus positive red cells, IgGl YB2 / 0 or the antibody IgG3 D29 expressed in YB2 / 0 or CHO.
- the secretion of different cytokines IL1 beta, IL6, IFN gamma, TNF alpha is measured in the supernatants after overnight incubation by ELISA technique.
- the levels of IL1 beta and of IFN gamma are identical for all the antibodies.
- the levels of IL6 produced by the monocytes are comparable for the IgG1 and IgG3 produced in YB2 / 0 but lower for the IgG3 produced in CHO.
- TNF alpha a slight decrease is observed in the presence of IgG3 produced by CHO.
- Example 8 Ability of anti-D IgG3 to induce phagocytosis of Rhesus positive red cells by THP-1 cells.
- Phagocytosis test THP-1 cells are incubated in the presence of Rhesus positive red cells and antibodies. The number of cells having phagocytosed at least one red blood cell is evaluated by counting under the microscope. The results are expressed as a percentage of cells having phagocytosed at least one red blood cell (see FIG. 10).
- the IgG of the anti-D polyclonal antibody WinRho have the strongest capacity (43.4%) to induce phagocytosis of Rhesus positive red cells by the THP-1 cell.
- IgGl YB2 / 0 is not very active (14.6%).
- IgG3 YB2 / 0 induces a phagocytosis of 34.5%, superior to purified polyclonal IgG3 (IgG3 WinRlio).
- the weakest phagocytosis activities are obtained with the IgG3 produced by the fused B lymphocyte (D29 P3 X229) and the IgG3 produced in CHO.
- the expression of an IgG3 in YB2 / 0 potentiates its capacities to induce phagocytosis, this property being able to be particularly interesting in infectious diseases as well as in Alzeimer's disease (McGeer PL, McGeer E. Immunotherapy for Alzheimer's disease, Sci Aging Knowledge Environ. 2004 Jul 07; 2004 ⁇ _and this in comparison with an IgG3 expressed in CHO or secreted by a heteromyeloma.
- the particular glycan profile of PIgG3 produced in YB2 / 0, that is to say, short forms, not sialylated, and with a fucose level of less than 35% gives it original properties as demonstrated above: - strong fixation CD 16 which is comparable to that of IgGl, potentiation of phagocytosis an increase in ADCC activity in the presence of PBMC or NK cells compared to IgG3 produced in CHO. an ability to compete with IgGl at the level of their binding to CD 16 and thus negatively modulate the secretion of cytokines.
Abstract
Description
Claims
Priority Applications (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2006534803A JP2007533639A (ja) | 2003-10-16 | 2004-10-18 | 強い食作用活性を有するYB2/0細胞株の抗RHESUS−DIgG3 |
EP04817229A EP1675616A2 (fr) | 2003-10-16 | 2004-10-18 | Nouvelles igg3 utiles pour stimuler la phagocytose |
US10/575,218 US20070218052A1 (en) | 2003-10-16 | 2004-10-18 | Novel lgG3 Antibodies for Stimulating Phagocytosis |
AU2004281220A AU2004281220A1 (en) | 2003-10-16 | 2004-10-18 | IGG3 anti rhesus-D in the line YB2/0 having a high phagocytosis activity |
CA002542508A CA2542508A1 (fr) | 2003-10-16 | 2004-10-18 | Igg3 anti rhesus-d dans la lignee yb2/0 ayant une forte activite en phagocytose |
BRPI0415420-7A BRPI0415420A (pt) | 2003-10-16 | 2004-10-18 | igg3 úteis para estimular a fagocitose |
IL174949A IL174949A0 (en) | 2003-10-16 | 2006-04-11 | ANTI RHESUS-D IgG3 IN A YB2/0 CELL LINE HAVING A STRONG PHAGOCYTOSIS ACTIVITY |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR0312087A FR2861078B1 (fr) | 2003-10-16 | 2003-10-16 | NOUVELLES IgG3 UTILES POUR STIMULER LA PHAGOCYTOSE |
FR0312087 | 2003-10-16 |
Publications (3)
Publication Number | Publication Date |
---|---|
WO2005037866A2 true WO2005037866A2 (fr) | 2005-04-28 |
WO2005037866A3 WO2005037866A3 (fr) | 2005-07-21 |
WO2005037866A8 WO2005037866A8 (fr) | 2006-06-01 |
Family
ID=34385205
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/FR2004/002657 WO2005037866A2 (fr) | 2003-10-16 | 2004-10-18 | Igg3 anti rhesus-d dans la lignee yb2/0 ayant une forte activite en phagocytose |
Country Status (9)
Country | Link |
---|---|
US (1) | US20070218052A1 (fr) |
EP (1) | EP1675616A2 (fr) |
JP (1) | JP2007533639A (fr) |
AU (1) | AU2004281220A1 (fr) |
BR (1) | BRPI0415420A (fr) |
CA (1) | CA2542508A1 (fr) |
FR (1) | FR2861078B1 (fr) |
IL (1) | IL174949A0 (fr) |
WO (1) | WO2005037866A2 (fr) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005053742A1 (fr) * | 2003-12-04 | 2005-06-16 | Kyowa Hakko Kogyo Co., Ltd. | Medicament contenant une composition a base d'anticorps |
FR2957598B1 (fr) | 2010-03-17 | 2015-12-04 | Lfb Biotechnologies | Nouveau peptide signal, et son utilisation pour la production de proteines recombinantes |
FR3003171B1 (fr) * | 2013-03-15 | 2015-04-10 | Lab Francais Du Fractionnement | Nouveaux medicaments comprenant une composition d'anticorps enrichie en isoforme de charge majoritaire |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5653978A (en) * | 1987-09-18 | 1997-08-05 | The National Blood Authority | Human anti-Rh(D) monoclonal antibodies and methods of use of antibodies in immunotherapy |
WO2001077181A2 (fr) * | 2000-04-12 | 2001-10-18 | Laboratoire Francais Du Fractionnement Et Des Biotechnologies | Anticorps monoclonaux anti-rhesus d |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
BR9305557A (pt) * | 1992-06-26 | 1997-03-25 | Aetsrn | Anticorpos monoclonais humanos anti Rhesus-D e composiçao farmacêutica que os contém |
DE10001372A1 (de) * | 2000-01-14 | 2001-08-02 | Deutsches Krebsforsch | Anti-CD3-Einzelketten-Antikörper mit humanem Cmu3- und Cmu4- Domänen |
US7605235B2 (en) * | 2003-05-30 | 2009-10-20 | Centocor, Inc. | Anti-tissue factor antibodies and compositions |
-
2003
- 2003-10-16 FR FR0312087A patent/FR2861078B1/fr not_active Expired - Fee Related
-
2004
- 2004-10-18 WO PCT/FR2004/002657 patent/WO2005037866A2/fr active Application Filing
- 2004-10-18 BR BRPI0415420-7A patent/BRPI0415420A/pt not_active IP Right Cessation
- 2004-10-18 CA CA002542508A patent/CA2542508A1/fr not_active Abandoned
- 2004-10-18 AU AU2004281220A patent/AU2004281220A1/en not_active Abandoned
- 2004-10-18 JP JP2006534803A patent/JP2007533639A/ja active Pending
- 2004-10-18 EP EP04817229A patent/EP1675616A2/fr not_active Withdrawn
- 2004-10-18 US US10/575,218 patent/US20070218052A1/en not_active Abandoned
-
2006
- 2006-04-11 IL IL174949A patent/IL174949A0/en unknown
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5653978A (en) * | 1987-09-18 | 1997-08-05 | The National Blood Authority | Human anti-Rh(D) monoclonal antibodies and methods of use of antibodies in immunotherapy |
WO2001077181A2 (fr) * | 2000-04-12 | 2001-10-18 | Laboratoire Francais Du Fractionnement Et Des Biotechnologies | Anticorps monoclonaux anti-rhesus d |
Non-Patent Citations (5)
Title |
---|
BREDIUS R G M ET AL: "ROLE OF NEUTROPHIL FCGRIIA (CD32) AND FCGRIIB (CD16) POLYMORPHIC FORMS IN PHAGOCYTOSIS OF HUMAN IGG1- AND IGG3-OPSONIZED BACTERIA AND ERYTHROCYTES" IMMUNOLOGY, OXFORD, GB, vol. 83, no. 4, 1994, pages 624-630, XP000914745 ISSN: 0019-2805 * |
KLEIN P ET AL: "HUMAN RECOMBINANT ANTI-RH(D) MONOCLONAL ANTIBODIES: IMPROVEMENT OF BILOGICAL PROPERTIES BY IN VITRO CLASS-SWITCH" HUMAN ANTIBODIES, AMSTERDAM, NL, vol. 8, no. 1, 1997, pages 17-25, XP001015726 ISSN: 1093-2607 * |
SCHARF O ET AL: "Immunoglobulin G3 from polyclonal human immunodeficiency virus (HIV) immune globulin is more potent than other subclasses in neutralizing HIV type 1" JOURNAL OF VIROLOGY, THE AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 75, no. 14, juillet 2001 (2001-07), pages 6558-6565, XP002219983 ISSN: 0022-538X * |
SHINKAWA T ET AL: "The absence of fucose but not the presence of galactose or bisecting N-acetylglucosamine of human IgG1 complex-type oligosaccharides shows the critical role of enhancing antibody-dependent cellular cytotoxicity" JOURNAL OF BIOLOGICAL CHEMISTRY, AMERICAN SOCIETY OF BIOLOGICAL CHEMISTS, BALTIMORE, MD, US, vol. 278, no. 5, 31 janvier 2003 (2003-01-31), pages 3466-3473, XP002965857 ISSN: 0021-9258 * |
SHITARA KENYA ET AL: "A new vector for the high level expression of chimeric antibodies in myeloma cells" JOURNAL OF IMMUNOLOGICAL METHODS, vol. 167, no. 1-2, 1994, pages 271-278, XP002911931 ISSN: 0022-1759 * |
Also Published As
Publication number | Publication date |
---|---|
WO2005037866A3 (fr) | 2005-07-21 |
FR2861078A1 (fr) | 2005-04-22 |
FR2861078B1 (fr) | 2007-09-21 |
US20070218052A1 (en) | 2007-09-20 |
IL174949A0 (en) | 2006-08-20 |
EP1675616A2 (fr) | 2006-07-05 |
AU2004281220A1 (en) | 2005-04-28 |
WO2005037866A8 (fr) | 2006-06-01 |
CA2542508A1 (fr) | 2005-04-28 |
JP2007533639A (ja) | 2007-11-22 |
BRPI0415420A (pt) | 2006-12-05 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP1537147B1 (fr) | Anticorps anti hla-dr ayant une adcc augmentée et induisant la production de cytokines. | |
TWI660972B (zh) | 抗mcam抗體及相關使用方法 | |
EP3159008B1 (fr) | Utilisation d'anticorps optimises en adcc pour traiter les patients faibles repondeurs | |
CA2498383C (fr) | Traitement des pathologies echappant a la reponse immune par des anticorps optimises | |
TWI359671B (en) | Cd40 antibody formulation and methods | |
KR101297146B1 (ko) | 인간화 FcγRIIB 특이적 항체 및 그의 사용 방법 | |
CN111727197A (zh) | 抗pd-1抗体和治疗方法 | |
KR20190117489A (ko) | 항-bcma 중쇄-단독 항체 | |
FR2915398A1 (fr) | "ensemble de moyens pour le traitement d'une pathologie maligne, d'une maladie auto-immune ou d'une maladie infectieuse" | |
WO2019115773A1 (fr) | Variants avec fragment fc ayant une affinité augmentée pour fcrn et une affinité augmentée pour au moins un récepteur du fragment fc | |
JP2022516301A (ja) | Cd137アゴニスト性抗体とその使用 | |
WO2005037866A2 (fr) | Igg3 anti rhesus-d dans la lignee yb2/0 ayant une forte activite en phagocytose | |
EP1537419B1 (fr) | Procédé d'évaluation de l'efficacité ADCC médiée par le CD16 d'anticorps monoclonaux ou polyclonaux | |
HUT77342A (hu) | Immunoszupresszív aktivitással rendelkező monoklonális antitest fragmensek | |
CA2542928A1 (fr) | Utilisation de cations metalliques pour cristalliser le fragment fc d'un anticorps rhesus d | |
WO2018109210A1 (fr) | Combinaison d'anticorps anti-cd303 et anti-her2 | |
FR2910896A1 (fr) | Anticorps monoclonal dirige contre le recepteur humain des ldl |
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 BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A2 Designated state(s): BW GH GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LU MC NL PL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG |
|
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: 174949 Country of ref document: IL Ref document number: 2004817229 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2542508 Country of ref document: CA Ref document number: 2004281220 Country of ref document: AU |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2006534803 Country of ref document: JP |
|
ENP | Entry into the national phase |
Ref document number: 2004281220 Country of ref document: AU Date of ref document: 20041018 Kind code of ref document: A |
|
WWP | Wipo information: published in national office |
Ref document number: 2004281220 Country of ref document: AU |
|
CFP | Corrected version of a pamphlet front page |
Free format text: UNDER (54) PUBLISHED TITLE IN ENGLISH REPLACED BY CORRECT TITLE AND UNDER (57) PUBLISHED ABSTRACT IN ENGLISH REPLACED BY CORRECT ABSTRACT |
|
WWP | Wipo information: published in national office |
Ref document number: 2004817229 Country of ref document: EP |
|
ENP | Entry into the national phase |
Ref document number: PI0415420 Country of ref document: BR |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2007218052 Country of ref document: US Ref document number: 10575218 Country of ref document: US |
|
WWP | Wipo information: published in national office |
Ref document number: 10575218 Country of ref document: US |