isscr摘要 ips产生单克隆T细胞

stemcell8

W 1 - MONOCLONAL GUIDANCE OF T LINEAGE LYMPHOCYTES FROM HUMAN INDUCED PLURIPOTENT STEM CELLS ORIGINATING FROM A SINGLE PERIPHERAL T LYMPHOCYTE' d! e2 B5 v$ F
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Nishimura, Toshinobu1, Kaneko, Shin1, Gotoh, Haruo1, Takayama, Naoya1, Nakamura, Sou1, Shimizu, Takafumi1, Iriguchi, Shoichi1, Tachikawa-Kawana, Ai2, Takahashi, Satoshi3, Watanabe, Nobukazu4, Otsu, Makoto1, Eto, Koji1, Nakauchi, Hiromitsu1
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4 z, V2 N+ m" H1Division of Stem Cell Therapy, Center for Stem Cell Biology and Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan, 2Division of Infectious Diseases, Advanced Clinical Research Center, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan, 3Division of Molecular Therapy, Advanced Clinical Research Center, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan, 4Laboratory of Diagnostic Medicine, Center for Stem Cell Biology and Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan9 M- H8 W, P  W0 K9 I

2 F& K6 o( Z7 T& M1 z4 n0 pT lymphocytes exert their effecter functions and play a central role in cellular immunity by recognizing antigens using T-cell receptors (TCRs) in an HLA-restricted, antigen-specific manner. There has been much effort to develop clinically relevant methods for using antigen- or disease-specific TCR expressing T lymphocytes purified and expanded in vitro. To date, however, such T-cells have not yet proved effective because they proceed to exhaustion; losing the potential for both long term survival and proliferation. To overcome this obstacle to T-cell-based immunotherapy, we endeavored to generate monoclonal T lymphocytes from induced pluripotent stem (iPS) cells derived from antigen-reactive T lymphocytes. iPS cells have a capacity for unlimited self-renew while maintaining pluripotency. This feature enabled us to induce an unlimited number of clonal T lymphocytes showing reactivity to specific antigens when iPS cells were established from antigen-specific T-cells. In addition, they may proliferate for a longer period than their peripheral blood counterparts expanded in vitro, if they retain properties of naive T-cells. TCR subunits are encoded by four genes (TCRA, TCRB, TCRG and TCRD) assembled and rearranged in a strictly ordered manner during the development of early T-cell progenitors into double positive thymocytes. Therefore, extensive and monoclonal induction of antigen-specific T lineage cells may be feasible, if TCR gene arrangements are conserved during the reprogramming step. It is also possible, however, that another allele or TCR gene family may be assembled during differentiation. Peripheral T lymphocytes were isolated from healthy volunteers, after which three, four or six reprogramming factors (OCT3/4, SOX2, c-MYC, KLF4, NANOG and LIN28) were transduced into fresh or thawed T lymphocytes using a retroviral vector system. The transduced T-cells were then cultured on mouse embryonic fibroblasts (MEFs) in the presence of cytokines and chemicals favorable for T-cell survival/proliferation, and iPSC-like colonies were observed within about 2 weeks. Colonies selected and clonally expanded exhibited standard ES-like morphology, cell surface markers and alkaline phosphatase activity, as well as differentiation into various tissues related to the three germ layers seen in teratomas. The TCR gene rearrangements engraved in these iPS colonies were determined, and shown to be monoclonal in all iPS lines. The reliable commitment into T lineage cells by murine stromal cell line (OP-9DL1) based coculture was evidenced by expression of TCRα/β heterodimer and cell surface markers such as CD3. Genetic and phenotypic characterization of their derivation showed that differentiation of T-cell-derived iPS (T-iPS) cells was directed toward the T lineage. Genetic analyses confirmed that the induced T lineage cells had TCRs identical to those engraved in the pre-differentiated T-iPS cells, and the two sequences were found to be completely identical copies at the mono-nucleotide level, even in the most highly diversifiable region of TCR chains (complementarity-determining region 3: CDR3). These data suggest that further optimization of these processes for T-iPS generation and monoclonal disease-specific T lineage cell expansion could lead to the development of indefinitely and repeatedly suppliable disease- and patient-specific T-cell therapy.6 g( L" r/ D+ Z3 G- A9 U" m

gact

要全文。。

stemcell8

没全文，就是个会议摘要