Adoptive transfer of T-cell precursors enhances T-cell reconstitution after allogeneic hematopoietic stem cell transplantation

Abstract

Immunoincompetence after allogeneic hematopoietic stem cell transplantation (HSCT) affects in particular the T-cell lineage and is associated with an increased risk for infections, graft failure and malignant relapse. To generate large numbers of T-cell precursors for adoptive therapy, we cultured mouse hematopoietic stem cells (HSCs) in vitro on OP9 mouse stromal cells expressing the Notch-1 ligand Delta-like-1 (OP9-DL1). We infused these cells, together with T-cell–depleted mouse bone marrow or purified HSCs, into lethally irradiated allogeneic recipients and determined their effect on T-cell reconstitution after transplantation. Recipients of OP9-DL1–derived T-cell precursors showed increased thymic cellularity and substantially improved donor T-cell chimerism (versus recipients of bone marrow or HSCs only). OP9-DL1–derived T-cell precursors gave rise to host-tolerant CD4+ and CD8+ populations with normal T-cell antigen receptor repertoires, cytokine secretion and proliferative responses to antigen. Administration of OP9-DL1–derived T-cell precursors increased resistance to infection with Listeria monocytogenes and mediated significant graft-versus-tumor (GVT) activity but not graft-versus-host disease (GVHD). We conclude that the adoptive transfer of OP9-DL1–derived T-cell precursors markedly enhances T-cell reconstitution after transplantation, resulting in GVT activity without GVHD.

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Figure 1: Coculture of HSCs on OP9-DL1 cells results in large numbers of T-cell precursors, including DN2 cells, that engraft in the thymus of allogeneic HSCT recipients.
Figure 2: OP9-DL1–derived T-cell precursors significantly improve thymic and peripheral T-cell reconstitution after allogeneic HSCT.
Figure 3: Combined T-cell precursor and KGF administration has additive effects on thymic reconstitution after allogeneic HSCT, and OP9-DL1–derived T-cell precursors enhance intestinal T-cell development in thymectomized mice after allogeneic HSCT.
Figure 4: OP9-DL1–derived T cells have a diverse TCR repertoire, give an intact proliferative response in a third-party MLR and increase their cytokine secretion upon stimulation.
Figure 5: Adoptive transfer of OP9-DL1–derived T-cell precursors enhances resistance to L. monocytogenes after allogeneic HSCT.
Figure 6: OP9-DL1–derived T-cell precursors do not induce GVHD but promote GVT.

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Acknowledgements

The authors would like to thank the staff of the Research Animal Resource Center for animal care. This work was supported by grants HL69929, CA33049 and CA107096 from the US National Institutes of Health, by awards from the Leukemia and Lymphoma Society the Emerald Foundation and The Experimental Therapeutics Center of Memorial Sloan-Kettering Cancer Center funded by Mr. William H. Goodwin and Mrs. Alice Goodwin and the Commonwealth Foundation for Cancer Research (M.R.M.v.d.B.), P20-CA103694 from the National Institutes of Health (Ö.A.), Dr. Werner Jackstaedt Stiftung (J.L.Z.), Deutsche Forschungsgemeinschaft (J.L.Z.), Deutscher Akademischer Austausch Dienst (T.H.T.), Boehringer Ingelheim Fonds (T.H.T.) and Deutsche Krebshilfe (T.D.K.). Ö.A. is the recipient of the Amy Strelzer Manasevit Scholar Award from The National Marrow Donor Program (NMDP) and The Marrow Foundation. J.C.Z.P. is supported by a Canada Research Chair in Developmental Immunology.

Author information

J.L.Z. designed experiments, performed animal studies and tissue culture, analyzed data, generated figures and wrote the manuscript. A.A.K, S.X.L., T.H.T., T.D.K, V.M.H., S.J.M., D.S, O.M.S, J.G., N.P., A.C., J.C.P., and R.R. assisted with performance of transplantations, HSC sorting, organ harvests, in vivo BLI, GVHD scoring and tissue culture. A.D. and M.A.P. designed and performed vaccination experiments and G.R. performed ELISpot studies. E.M. assisted with L. monocytogenes studies and E.G.P. designed and analyzed L. monocytogenes experiments. G.H. performed statistical analyses and J.C.Z.-P. designed experiments and analyzed data. Ö.A. designed and performed experiments, analyzed data and supervised the study, and M.R.M.v.d.B. designed experiments, analyzed data, supervised the study and edited the manuscript.

Correspondence to Marcel R M van den Brink.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

OP9-DL1 derived DN T-cell precursors upregulate several molecules involved in lymphocyte trafficking (PDF 501 kb)

Supplementary Fig. 2

Thymic engraftment of OP9-DL1 derived T-cell precursors after allogeneic HSCT is dose and time point dependent, and its effect on thymic reconstitution cannot be resembled by increasing the stem cell inoculum in the allograft (PDF 208 kb)

Supplementary Fig. 3

Adoptive transfer of OP9-DL1 derived precursor cells enhances early NK cell reconstitution in recipients of an allogeneic HSCT (PDF 129 kb)

Supplementary Fig. 4

T-cells from tumor bearing BMT recipients treated with OP9-DL1 derived T-cell precursors show an increased INF-γ response to stimulation with A20-TGL (PDF 100 kb)

Supplementary Fig. 5

NK cell are not required for GVT activity against A20-TGL mediated by OP9-DL1 derived precursor cells in allogeneic HSCT recipients (PDF 136 kb)

Supplementary Methods (PDF 123 kb)

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