Tumor immunotherapy across MHC barriers using allogeneic T-cell precursors

Abstract

We present a strategy for adoptive immunotherapy using T-lineage committed lymphoid precursor cells generated by Notch1-based culture. We found that allogeneic T-cell precursors can be transferred to irradiated individuals irrespective of major histocompatibility complex (MHC) disparities and give rise to host-MHC restricted and host-tolerant functional allogeneic T cells, improving survival in irradiated recipients as well as enhancing anti-tumor responses. T-cell precursors transduced to express a chimeric receptor targeting hCD19 resulted in significant additional anti-tumor activity, demonstrating the feasibility of genetic engineering of these cells. We conclude that ex vivo generated MHC-disparate T-cell precursors from any donor can be used universally for 'off-the-shelf' immunotherapy, and can be further enhanced by genetic engineering for targeted immunotherapy.

Stable gene transfer and expression in cord blood–derived CD34+ hematopoietic stem and progenitor cells by a hyperactive Sleeping Beauty transposon system

“…3,4 Gene therapy targeting HSCs has shown promise in the treatment of HIV infections and cancer. [5][6][7][8] The potential of HSC-based gene therapy is enormous as HSCs are able to self-renew and undergo differentiation into progenitor populations, ultimately leading to the generation of mature cells of multiple lineages with diverse functions. Therefore, ex vivo stable gene transfer into HSCs followed by transplantation could result in the long-term persistence of genetically modified HSCs in the recipient, providing a potential cure to a number of disorders affecting components of the hematopoietic system.…”

Extrathymic development of murine T cells after bone marrow transplantation

“…Having observed extrathymic development of BM-derived T cell progenitors after BMT, we sought to determine whether preTs also underwent extrathymic development (16,17). We generated preTs by coculturing lineage -c-kit + Sca-1 + hematopoietic progenitors on the OP9-DL1 stromal cell layer for up to 26 days in the presence of IL-7 and Flt3L.…”

“…We have previously demonstrated that adoptive transfer of T cell precursors (preTs) generated ex vivo on the OP9-DL1 system into BMT recipients significantly increased thymopoiesis, circulating T cell numbers, and protection against bacterial infection (16,17). While this is primarily dependent on improved thymopoiesis, based on studies in thoracically thymectomized BMT recipients, we hypothesized that extrathymic T cell development could contribute to the regeneration of the posttransplant T cell pool (16).…”

“…Expansion of peripheral T cells after BMT can lead to a pool of memory phenotype T cells with limited T cell receptor diversity (11, 12), leading to decreased immune function as well as poorer prognosis for older BMT patients (13-15). Consequently, mechanisms to circumvent the thymus to generate functional T cells could drastically improve not only BMT outcomes, but outcomes for all immunodeficient patients.We have previously demonstrated that adoptive transfer of T cell precursors (preTs) generated ex vivo on the OP9-DL1 system into BMT recipients significantly increased thymopoiesis, circulating T cell numbers, and protection against bacterial infection (16,17). While this is primarily dependent on improved thymopoiesis, based on studies in thoracically thymectomized BMT recipients, we hypothesized that extrathymic T cell development could contribute to the regeneration of the posttransplant T cell pool (16).…”

Concurrent visualization of trafficking, expansion, and activation of T lymphocytes and T-cell precursors in vivo

“…10,17 Here we used these ex vivo-generated T-cell precursors and our dual in vivo imaging system to assess T-cell development and selection during immune reconstitution. We adoptively transferred CD4 Ϫ CD8 Ϫ CD25 ϩ CD44 ϩ (DN2) and CD4 Ϫ CD8 Ϫ CD25 ϩ CD44 Ϫ (DN3) T cell precursor cells derived from an OP9-DL1 coculture of transduced LSK cells into lethally irradiated recipients of allogeneic lineagedepleted BM cells.…”