Anti third party central memory t cells, methods of producing same and use of same in transplantation and disease treatment

Inactive Publication Date: 2014-07-31
YEDA RES & DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides a method for generating a population of cells with a central memory T-lymphocyte (Tcm) phenotype, which are tolerance-inducing cells and / or endowed with anti-disease activity, that can homing to lymph nodes after transplantation. The method involves contacting peripheral blood mononuclear cells (PBMC) with a third party antigen or antigens in the presence of IL-21 to enrich antigen reactive cells, and then culturing these cells in an antigen-free environment with IL-21, IL-15, and IL-7 to allow proliferation of cells with the Tcm phenotype.

Problems solved by technology

However, in leukemia patients, the clinical outcome of T cell depleted BM was disappointing, as the benefit of GVHD prevention was offset by a markedly increased rate of graft rejection.
Nevertheless, this strategy is hampered by opportunistic infections due to slow immune reconstitution and considerable toxicities of the immunosuppressants.
While in high risk leukemia patients such transplant-related mortality can be acceptable, it would be intolerable if applied to patients with a long life expectancy.
Yet, the marked levels of host immune cells surviving the mild preparatory regimens, represents a difficult barrier for the engraftment of donor cells.
However, a significant increase in the BM inoculum has been difficult to achieve in humans.

Method used

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  • Anti third party central memory t cells, methods of producing same and use of same in transplantation and disease treatment
  • Anti third party central memory t cells, methods of producing same and use of same in transplantation and disease treatment
  • Anti third party central memory t cells, methods of producing same and use of same in transplantation and disease treatment

Examples

Experimental program
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Effect test

example 1

Generation and Optimization of Human Anti-Third Party T Central Memory (Tcm) Cells

[0272]In order to translate the mouse studies previously presented to clinical application, the procedure was optimized for generating human anti-3rd party cytotoxic T lymphocytes (CTLs). To that end, different parameters were evaluated including different reagents for the isolation of CD8 responder cells, the composition of the stimulators and the cytokine milieu.

[0273]Potentially, as found in the previously presented mouse model, treatment with central memory T cells (Tcm) could be valuable either in the context of autologous [Lask A et al., Blood (ASH Annual Meeting Abstracts), (2010) 116: 424] or in allogeneic bone marrow transplant (BMT) [Ophir E et al., Blood. (2010) 115(10): 2095-104; Ophir E., 37th EBMT annual meeting, Apr. 3-6, 2011, Paris, France. Oral Presentation Abstract Nr: 662].

[0274]In the human autologous setting (FIG. 1A) anti-3rd party Tcm can be administrated together with autologou...

example 2

Large Scale Preparation of Human Anti-3Rd Party Tcm in Plastic Bags Using GMP Grade Reagents

[0316]To simulate the conditions anticipated when using patients own PBMC for the generation of autologous Tcm, initially two large scale leukaphersis procedures were performed, from two normal donors, and a large number of mononuclear cells were cryopreserved (divided into several batches). Each batch was used for one large scale experiment. In the first experiment, several technical problems were encountered including difficulty in the generation of DCs from the frozen bags according to the Wurzburg protocol and sourcing of a new GMP grade IL-15 for which the biological activity was unclear. These problems, which resulted in a very poor CD8 T cell expansion (around 3 folds), were corrected in the subsequent experiments by using the presently described protocol for DCs (as described above) and by using the appropriate concentration of IL-15 (i.e. 300 U / ml).

[0317]As can be seen in FIG. 15, wh...

example 3

GVL Potential of the Human Anti-3rd Party Tcm Against an Established Cell Line

[0319]Considering that in the autologous setting the potential use of anti-3rd party Tcm is solely for eradicating residual tumor cells (in the allogeneic setting it also serves to enhance engraftment of the BM cells) it is important to develop a straightforward assay for cytotoxic capacity ex-vivo, which could be used for quality control prior to infusion of the Tcm to the patient. To that end, a TCR independent assay was used based on the demonstration by Lask et al. [Lask A et al., J. Immunol. (2011) 187(4):2006-14] that a MHC mutated line not recognizable by TCR due to this mutation, can still be killed by anti-3rd party CTLs through their TCR independent killing mechanism. Clearly, if applicable also for human Tcm, such a killing modality could serve to distinguish the Tcm killing from that exhibited by NK cells.

[0320]To address this question, a mixed lymphocyte reaction (MLR) was carried out with ant...

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Abstract

A method of generating an isolated population of cells comprising anti-third party cells having a central memory T-lymphocyte (Tcm) phenotype, the cells being tolerance-inducing cells and / or endowed with anti-disease activity, and capable of homing to the lymph nodes following transplantation is disclosed. The method comprising: (a) contacting peripheral blood mononuclear cells (PBMC) with a third party antigen or antigens in the presence of IL-21 so as to allow enrichment of antigen reactive cells; and (b) culturing the cells resulting from step (a) in the presence of IL-21, IL-15 and IL-7 in an antigen free environment so as to allow proliferation of cells comprising the central memory T-lymphocyte (Tcm) phenotype.

Description

FIELD AND BACKGROUND OF THE INVENTION[0001]The present invention, in some embodiments thereof, relates to tolerance inducing and / or graft versus leukemia reactive anti-third party cells comprising central memory T-lymphocyte phenotype and, more particularly, but not exclusively, to methods of generating same and to the use of same in transplantation and in disease treatment.[0002]Bone marrow (BM) transplantation offers a curative treatment for many patients with hematological malignancies and other hematological disorders. However, the BM graft contains donor T cells which respond to the host antigens (Ags) and cause multi-system graft-versus-host disease (GVHD). In the early 80's bone marrow transplant (BMT), without the deleterious effect of GVHD, was demonstrated in the haploidentical (three HLA loci mismatched) settings, in severe combined immunodeficiency (SCID) patients. The problem of GVHD, which is almost uniformly lethal in such settings, was completely prevented by T cell ...

Claims

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Application Information

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IPC IPC(8): A61K35/14A61K35/28A61K35/407A61K35/39A61K35/26A61K35/34A61K35/42A61K35/36A61K35/38C12N5/0783A61K35/22A61K35/12A61K39/00
CPCA61K35/17C12N5/0636A61K35/28A61K35/407A61K35/39A61K35/22A61K2035/122A61K35/42A61K35/36A61K35/38A61K35/26A61K2035/124A61K35/34A61K39/001C12N5/0637C12N2501/2307C12N2501/2315C12N2501/2321C12N2502/1121A61P31/12A61P35/00A61P35/02A61P37/02A61K2239/26A61K39/46434A61K39/461A61K39/4621A61K39/4611A61K2039/5158
Inventor REISNER, YAIREIDELSTEIN, YAKIOPHIR, ERANLASK, ASSAFAFIK, RANOR-GEVA, NOGABACHAR-LUSTIG, ESTHER
Owner YEDA RES & DEV CO LTD
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