Method for proliferation of antigen-specific T cells
Inventors
Karlsson-Parra, Alex • Wallgren, Anna-Carin • Andersson, Bengt
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-9789174-B2
Publication Date
2017-10-17
Expiration Date
Abstract
The present invention relates to an in vitro method for priming genetically modified T cells suitable for administration to a patient having a tumor. The invention is also directed to the composition obtained by the method and uses thereof.
Core Innovation
The invention relates to antigen specific CD4+ and/or CD8+ T cells obtained by in vitro priming of genetically modified antigen specific CD4+ and/or CD8+ T cells suitable for administration to a patient having a tumor. The priming comprises co-culturing tumor antigen receptor expressing target T cells from the patient to be treated, mature dendritic cells, anti-CD3 antibodies and lymphocytes sensitized against MHC class I and/or MHC class II antigens on antigen presenting cells (APCs).
The priming uses allo-sensitized allogeneic lymphocytes (ASALs) generated by mixed leukocyte reaction and sensitized for MHC class I and/or MHC class II. During priming, the sensitized lymphocytes act to upregulate DC CD70 and drive cytokine production including IL-12, IFN-γ and IL-2, enriching CD27+ CD8+ T cells and reducing apoptosis as indicated by lower Annexin-V. This supports enhanced proliferation upon restimulation.
In disclosed experimental support, the primed antigen-specific T cells show increased proliferative capacity after restimulation and reduced apoptotic death. They further show improved resistance to apoptosis-inducing H2O2 and doxorubicin, preserved killing with comparable expansion of CAR (e.g., GD2 CAR) compared to a standard REP approach, higher CD8:CD4 expansion, and improved resistance to tumor suppressive factors including IL-10 and TGF-β (± H2O2). The document further characterizes a culture process involving dendritic cell maturation and inflammatory/TLR3 stimulation such as poly-I:C and/or other TLR ligands, together with compositions/medicaments for tumor treatment.
Claims Coverage
The partial content provides two independent claims. Across these claims, the inventive features include the specific in vitro priming co-culture components and sensitization criteria for APC-presented MHC class I and/or MHC class II antigens, and the post-priming cellular suitability/quality constraints for patient administration, including proliferation ability, Annexin-V staining threshold, and/or surface expression of CD27 and/or CD28.
In vitro priming co-culture with MHC-sensitized APC lymphocytes
Antigen specific CD4+ and/or CD8+ T cells obtained by in vitro priming of genetically modified antigen specific CD4+ and/or CD8+ T cells, wherein the priming comprises co-culturing tumor antigen receptor expressing target T cells from the patient to be treated, mature dendritic cells, anti-CD3 antibodies and lymphocytes sensitized against MHC class I and/or MHC class II antigens on antigen presenting cells (APCs).
Patient-administration cell state constraints including Annexin-V and CD27/CD28
Antigen specific CD4+ and/or CD8+ T cells suitable for administration to a patient wherein said CD4+ and/or CD8+ T cells have the ability to proliferate, no more than 40% of the cells should exhibit positive staining for Annexin-V, and/or express CD27 and/or CD28 at their cell surface.
Together, the independent claims define antigen-specific CD4+ and/or CD8+ T cell products generated by a defined in vitro priming co-culture including mature dendritic cells, anti-CD3 antibodies, patient tumor antigen receptor target T cells, and MHC class I and/or MHC class II-sensitized lymphocytes on APCs, and further restrict the administered cells by proliferation ability and by specified viability/phenotype criteria (Annexin-V threshold and/or CD27/CD28 surface expression).
Stated Advantages
Enhanced proliferation on restimulation.
Reduced apoptotic death as indicated by lower Annexin-V positivity.
Improved resistance to apoptosis-inducing H2O2 and doxorubicin.
Preserved killing.
Comparable CAR expansion to a standard REP approach while showing higher CD8:CD4 expansion.
Improved resistance to tumor suppressive factors including IL-10 and TGF-β (± H2O2).
Documented Applications
Administration to a patient having a tumor using the disclosed antigen specific CD4+ and/or CD8+ T cell preparation (medicament) for tumor treatment.
Interested in licensing this patent?