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Anti-cd277 antibodies and uses thereof

a technology of anti-cd277 and antibodies, which is applied in the field of anti-cd277 antibodies, can solve the problems of not being able to overcome the immunosuppressive mechanisms observed in cancer patients, and no satisfactory approach has been proven to induce potent immune responses against vaccines, etc., and achieves the effect of increasing activity and/or survival

Inactive Publication Date: 2015-12-10
INST NAT DE LA SANTE & DE LA RECHERCHE MEDICALE (INSERM) +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0012]The present invention relates to an anti-CD277 antibody, which activates or inhibits the cytolytic function, cytokine production and proliferation of Vγ9 / Vδ2 T cells. The present invention also relates to an anti-CD277 antibody, which costimulates T cells together with CD3-TCR, and / or which costimulates T cells in addition to CD28-B7 costimulation. The present invention also relates to an anti-CD277 antibody, which increases the activity and / or survival of monocytes and dendritic cells.

Problems solved by technology

To date, no satisfactory approach has been proven to induce potent immune responses against vaccines, especially in cancer patients.
Methods have yet to be devised to overcome the immunosuppressive mechanisms observed in cancer patients, and during chronic infections.
Treatment of autoimmune diseases and prevention of transplantation rejection in graft versus host diseases (GVHD) depends on immunosuppressive agents that have serious side effects, or are not always effective.

Method used

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  • Anti-cd277 antibodies and uses thereof
  • Anti-cd277 antibodies and uses thereof
  • Anti-cd277 antibodies and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

Materials and Methods

[0220]Cell Cultures

[0221]Blood samples were obtained from 5 healthy volunteers enrolled among the staff members, after informed consent. Monocytes were separated from peripheral blood mononuclear cells, and isolated using the MACS CD14 isolation kit (Miltenyi Biotech, Auburn, Calif. USA). Monocytes were cultured for 5 days with 200 ng / ml of recombinant human GM-CSF and 10 ng / ml recombinant human IL-4 (Schering-Plough Research Institute). These cells were termed immature dendritic cells (iDC). To achieve the stimulation, monocytes and iDC were cultured with 10 ng / ml of LPS, ligand for TLR4, 30 μg / ml of R848 (Sigma-Aldrich, Milano, Italy), ligand for TLR7 / 8 and with 2.5 mg / ml of poly (I:C) (Sigma-Aldrich), ligand for TLR3, and were harvested after 48 h. The cells were cultured in RPMI 1640 supplemented with 10% heat-inactivated FCS, 5 mM L-glutamine and 50 IU / ml penicillin-streptomycin (from here on referred to as complete medium).

[0222]Flow Cytometry and Antibodi...

example 2

Materials and Methods

Cell Preparation

[0240]Peripheral blood mononuclear cells (PBMCs) were obtained from healthy volunteer donors provided by the “Etablissement Français du Sang” (EFS-Marseille-France) and isolated by fractionation over a density gradient of Lymphoprep© (Abcys). Human CD4+ T cells were negatively selected from isolated PBMCs by depletion of non-CD4+ T cells with magnetic beads using the T cell isolation kit II from Miltenyi Biotec®. Isolated CD4 cells were used for further experiments when purity was superior to 90%.

Generation of Monoclonal Antibodies (mAbs)

[0241]The mouse anti-human programmed death-1 (PD-1) mAB and three different clones of mouse anti-human CD277 were purified from ascites in our laboratory: Anti-PD-1 (clone PD1 3.1 with an IgG1 isotype) (ghiotto et al. Int Immunol., 2010), Anti-CD277 (clone 103.2 with an IgG2a isotype, clones 20.1 and 7.2 both with an IgG1 isotype) (compte et al.). The anti-CD277 (clone 20.1) mAb was labelled with Alexa Fluor 64...

example 3

Materials and Methods

Antibodies and Fab Fragmentation

[0279]Anti-CD277: anti-BT3-20.1 and 103.2 mAb were generated and validated as previously described [10]. Fab fragments of anti-BT3-20.1 were generated and purified with the Immunopure Fab Preparation Kit following the manufacturer's recommendation (Pierce). Protein purity was assessed by nonreducing SDS-PAGE.

Construction of Phylogenetic Trees

[0280]Phylogenetic analyses were performed using the automated genomic annotation platform FIGENIX (FIGENIX Annotation Platform: [http: / / figenix2.up.univmrs.fr / Figenix / index.jsp]) to retrieve sequences and alignments and perform phylogenetic reconstruction. The pipeline used applied three different methods of phylogenetic tree reconstruction, i.e. Maximum Parsimony [38], Maximum likelihood [39] and Neighbour Joining [40], and a midpoint rooted consensus tree was built. Bootstrapping was carried out with 1000 replications. Bootstrap values are reported for each method (for a detailed descriptio...

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Abstract

Described herein are anti-CD277 antibodies which: activates or inhibit the cytolytic function of Vγ9/Vδ2 T cells, and/or costimulates T cells together with CD3-TCR, and/or costimulates T cells in addition to CD28-B7 costimulation, and/or increases the activity and/or survival of monocytes and dendritic cells. The use of said antibodies in therapy is also described.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]The present application is a continuation-in-part of U.S. patent application Ser. No. 13 / 994,277 filed Sep. 24, 2013, which is a 371 national stage application based on PCT / EP2011 / 072787 filed Dec. 14, 2011, which claims priority to International Patent Application No. PCT / IB2010 / 003417 filed on Dec. 15, 2010, which is incorporated herein by reference in its entirety.FIELD OF THE INVENTION[0002]The invention relates to anti-CD277 antibodies and uses thereof.BACKGROUND OF THE INVENTION[0003]White blood cells are cells of the immune system involved in defending the body against pathogens. Among these cells, lymphocytes, monocytes, and dendritic cells can be cited. Monocytes may migrate from the bloodstream to other tissues and differentiate into tissue resident macrophages or dendritic cells. Dendritic cells play a role as antigen presenting cells (APC) that activate lymphocytes. Among lymphocytes, T cells can be divided into γδ T cells and...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07K16/28A61K39/00A61K39/395
CPCC07K16/2896A61K2039/505A61K39/0011A61K39/3955A61K39/00A61K39/001129C07K16/2818C07K16/2827C07K2317/55C07K2317/74C07K2317/75
Inventor OLIVE, DANIELBONNEVILLE, MARCSCOTET, EMMANUELHARLY, CHRISTELLEGUILLAUME, YVES
Owner INST NAT DE LA SANTE & DE LA RECHERCHE MEDICALE (INSERM)
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