Human CD8 T cells generated in vitro from hematopoietic stem cells are functionally mature

BMC Immunology - Tập 12 Số 1 - 2011
Génève Awong1, Elaine Herer2, Ross N. La Motte-Mohs1, Juan Carlos Zúñiga‐Pflücker1
1Department of Immunology, University of Toronto, and Sunnybrook Research Institute, 2075, Bayview Avenue, Toronto, Ontario, M4N 3M5, Canada
2Sunnybrook Health Sciences Centre, Women & Babies Program, 2075, Bayview Avenue, Toronto, Ontario, M4N 3M5, Canada

Tóm tắt

AbstractBackgroundT cell development occurs within the highly specialized thymus. Cytotoxic CD8 T cells are critical in adaptive immunity by targeting virally infected or tumor cells. In this study, we addressed whether functional CD8 T cells can be generated fullyin vitrousing human umbilical cord blood (UCB) hematopoietic stem cells (HSCs) in coculture with OP9-DL1 cells.ResultsHSC/OP9-DL1 cocultures supported the differentiation of CD8 T cells, which were TCR/CD3hiCD27hiCD1anegand thus phenotypically resembled mature functional CD8 single positive thymocytes. Thesein vitro-generated T cells also appeared to be conventional CD8 cells, as they expressed high levels ofEomesand low levels ofPlzf, albeit not identical toex vivoUCB CD8 T cells. Consistent with the phenotypic and molecular characterization, upon TCR-stimulation,in vitro-generated CD8 T cells proliferated, expressed activation markers (MHC-II, CD25, CD38), secreted IFN-γ and expressed Granzyme B, a cytotoxic T-cell effector molecule.ConclusionTaken together, the ability to direct human hematopoietic stem cell or T-progenitor cells towards a mature functional phenotype raises the possibility of establishing cell-based treatments for T-immunodeficiencies by rapidly restoring CD8 effector function, thereby mitigating the risks associated with opportunistic infections.

Từ khóa


Tài liệu tham khảo

Spits H: Development of alphabeta T cells in the human thymus. Nat Rev Immunol. 2002, 2: 760-772. 10.1038/nri913.

Storek J, Gooley T, Witherspoon RP, Sullivan KM, Storb R: Infectious morbidity in long-term survivors of allogeneic marrow transplantation is associated with low CD4 T cell counts. Am J Hematol. 1997, 54: 131-138. 10.1002/(SICI)1096-8652(199702)54:2<131::AID-AJH6>3.0.CO;2-Y.

Douek DC, Vescio RA, Betts MR, Brenchley JM, Hill BJ, Zhang L, Berenson JR, Collins RH, Koup RA: Assessment of thymic output in adults after haematopoietic stem-cell transplantation and prediction of T-cell reconstitution. Lancet. 2000, 355: 1875-1881. 10.1016/S0140-6736(00)02293-5.

van den Brink MR, Alpdogan O, Boyd RL: Strategies to enhance T-cell reconstitution in immunocompromised patients. Nat Rev Immunol. 2004, 4: 856-867. 10.1038/nri1484.

Schmitt TM, Zúñiga-Pflücker JC: Induction of T cell development from hematopoietic progenitor cells by delta-like-1 in vitro. Immunity. 2002, 17: 749-756. 10.1016/S1074-7613(02)00474-0.

Schmitt TM, de Pooter RF, Gronski MA, Cho SK, Ohashi PS, Zúñiga-Pflücker JC: Induction of T cell development and establishment of T cell competence from embryonic stem cells differentiated in vitro. Nat Immunol. 2004, 5: 410-417. 10.1038/ni1055.

La Motte-Mohs RN, Herer E, Zúñiga-Pflücker JC: Induction of T-cell development from human cord blood hematopoietic stem cells by Delta-like 1 in vitro. Blood. 2005, 105: 1431-1439. 10.1182/blood-2004-04-1293.

Awong G, Herer E, Surh CD, Dick JE, La Motte-Mohs RN, Zuniga-Pflucker JC: Characterization in vitro and engraftment potential in vivo of human progenitor T cells generated from hematopoietic stem cells. Blood. 2009, 114: 972-982. 10.1182/blood-2008-10-187013.

De Smedt M, Hoebeke I, Plum J: Human bone marrow CD34+ progenitor cells mature to T cells on OP9-DL1 stromal cell line without thymus microenvironment. Blood Cells Mol Dis. 2004, 33: 227-232. 10.1016/j.bcmd.2004.08.007.

Zakrzewski JL, Kochman AA, Lu SX, Terwey TH, Kim TD, Hubbard VM, Muriglan SJ, Suh D, Smith OM, Grubin J, et al: Adoptive transfer of T-cell precursors enhances T-cell reconstitution after allogeneic hematopoietic stem cell transplantation. Nat Med. 2006, 12: 1039-1047. 10.1038/nm1463.

Van Coppernolle S, Verstichel G, Timmermans F, Velghe I, Vermijlen D, De Smedt M, Leclercq G, Plum J, Taghon T, Vandekerckhove B, Kerre T: Functionally mature CD4 and CD8 TCRalphabeta cells are generated in OP9-DL1 cultures from human CD34+ hematopoietic cells. J Immunol. 2009, 183: 4859-4870. 10.4049/jimmunol.0900714.

Zhao Y, Parkhurst MR, Zheng Z, Cohen CJ, Riley JP, Gattinoni L, Restifo NP, Rosenberg SA, Morgan RA: Extrathymic generation of tumor-specific T cells from genetically engineered human hematopoietic stem cells via Notch signaling. Cancer Res. 2007, 67: 2425-2429. 10.1158/0008-5472.CAN-06-3977.

van Lent AU, Nagasawa M, van Loenen MM, Schotte R, Schumacher TN, Heemskerk MH, Spits H, Legrand N: Functional human antigen-specific T cells produced in vitro using retroviral T cell receptor transfer into hematopoietic progenitors. J Immunol. 2007, 179: 4959-4968.

Res P, Spits H: Developmental stages in the human thymus. Semin Immunol. 1999, 11: 39-46. 10.1006/smim.1998.0152.

Vanhecke D, Leclercq G, Plum J, Vandekerckhove B: Characterization of distinct stages during the differentiation of human CD69+CD3+ thymocytes and identification of thymic emigrants. J Immunol. 1995, 155: 1862-1872.

Res P, Blom B, Hori T, Weijer K, Spits H: Downregulation of CD1 marks acquisition of functional maturation of human thymocytes and defines a control point in late stages of human T cell development. J Exp Med. 1997, 185: 141-151. 10.1084/jem.185.1.141.

He X, Park K, Wang H, Zhang Y, Hua X, Li Y, Kappes DJ: CD4-CD8 lineage commitment is regulated by a silencer element at the ThPOK transcription-factor locus. Immunity. 2008, 28: 346-358. 10.1016/j.immuni.2008.02.006.

Pearce EL, Mullen AC, Martins GA, Krawczyk CM, Hutchins AS, Zediak VP, Banica M, DiCioccio CB, Gross DA, Mao CA, et al: Control of effector CD8+ T cell function by the transcription factor Eomesodermin. Science. 2003, 302: 1041-1043. 10.1126/science.1090148.

Narayanan S, Silva R, Peruzzi G, Alvarez Y, Simhadri VR, Debell K, Coligan JE, Borrego F: Human Th1 cells that express CD300a are polyfunctional and after stimulation up-regulate the T-box transcription factor eomesodermin. PLoS One. 2010, 5: e10636-10.1371/journal.pone.0010636.

Savage AK, Constantinides MG, Han J, Picard D, Martin E, Li B, Lantz O, Bendelac A: The transcription factor PLZF directs the effector program of the NKT cell lineage. Immunity. 2008, 29: 391-403. 10.1016/j.immuni.2008.07.011.

Rosenberg SA, Restifo NP, Yang JC, Morgan RA, Dudley ME: Adoptive cell transfer: a clinical path to effective cancer immunotherapy. Nat Rev Cancer. 2008, 8: 299-308. 10.1038/nrc2355.

June CH: Principles of adoptive T cell cancer therapy. J Clin Invest. 2007, 117: 1204-1212. 10.1172/JCI31446.

Kim JV, Latouche JB, Riviere I, Sadelain M: The ABCs of artificial antigen presentation. Nat Biotechnol. 2004, 22: 403-410. 10.1038/nbt955.

Delaney C, Heimfeld S, Brashem-Stein C, Voorhies H, Manger RL, Bernstein ID: Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution. Nat Med. 2010, 16: 232-236. 10.1038/nm.2080.

Watts KL, Delaney C, Humphries RK, Bernstein ID, Kiem HP: Combination of HOXB4 and Delta-1 ligand improves expansion of cord blood cells. Blood. 116: 5859-5866. 10.1182/blood-2010-05-286062.

Irwin MJ, Heath WR, Sherman LA: Species-restricted interactions between CD8 and the alpha 3 domain of class I influence the magnitude of the xenogeneic response. J Exp Med. 1989, 170: 1091-1101. 10.1084/jem.170.4.1091.

Schwartzberg PL, Mueller KL, Qi H, Cannons JL: SLAM receptors and SAP influence lymphocyte interactions, development and function. Nat Rev Immunol. 2009, 9: 39-46. 10.1038/nri2456.

Alonzo ES, Gottschalk RA, Das J, Egawa T, Hobbs RM, Pandolfi PP, Pereira P, Nichols KE, Koretzky GA, Jordan MS, Sant'Angelo DB: Development of promyelocytic zinc finger and ThPOK-expressing innate gamma delta T cells is controlled by strength of TCR signaling and Id3. J Immunol. 184: 1268-1279. 10.4049/jimmunol.0903218.

Ko HS, Fu SM, Winchester RJ, Yu DT, Kunkel HG: Ia determinants on stimulated human T lymphocytes. Occurrence on mitogen- and antigen-activated T cells. J Exp Med. 1979, 150: 246-255. 10.1084/jem.150.2.246.

Holling TM, van der Stoep N, Quinten E, van den Elsen PJ: Activated human T cells accomplish MHC class II expression through T cell-specific occupation of class II transactivator promoter III. J Immunol. 2002, 168: 763-770.

van Baarle D, Kostense S, van Oers MH, Hamann D, Miedema F: Failing immune control as a result of impaired CD8+ T-cell maturation: CD27 might provide a clue. Trends Immunol. 2002, 23: 586-591. 10.1016/S1471-4906(02)02326-8.

Hamann D, Baars PA, Rep MH, Hooibrink B, Kerkhof-Garde SR, Klein MR, van Lier RA: Phenotypic and functional separation of memory and effector human CD8+ T cells. J Exp Med. 1997, 186: 1407-1418. 10.1084/jem.186.9.1407.

Awong G, Motte-Mohs RN, Zúñiga-Pflücker JC: In vitro human T cell development directed by notch-ligand interactions. Methods Mol Biol. 2008, 430: 135-142. full_text.

Thông tin
Thông tin xuất bản

BMC Immunology

Tập 12 Số 1

Thông tin tác giả