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Induction of Graft-versus-host Disease and In Vivo T Cell Monitoring Using an MHC-matched Murine Model
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Tolerance induction across Mls and minor histocompatibility complex by inhibiting activation of T helper type 1 in

K Sugiura1, S Lee, T Nagahama

  • 1First Department of Pathology, Kansai Medical University, 10-15 Fumizono-cho, Moriguchi City, Osaka 570-8506, Japan.

Immunology Letters
|May 12, 2001
PubMed

We have previously succeeded in inducing persistent donor-specific tolerance across Mls plus multiple minor histocompatibility barriers by portal venous (p.v.) injection of donor spleen or bone marrow cells plus cyclophosphamide (CY) treatment. Microchimerism was established in the lymph-hemopoietic organs of the tolerant recipients. However, the mechanisms, particularly the roles of CY in the tolerance induction, have not been clarified. We examined the tolerance induction using other anti-mitotic agents and evaluated the in vitro proliferative responses and cytokine expression of T cells from the recipients after stimulation with donor alloantigens. The administration of not only CY but also mitomycin C (MMC) and cytosin arabinoside (Ara C) elicited a prolongation of skin graft survival. CY induced tolerance when it was administered 2 days after the p.v. injection, but not immediately or 4 days after the p.v. injection. T cells collected from the tolerant recipients showed no proliferative responses as a result of stimulation with donor alloantigens whereas the responses of T cells from non-tolerant recipients were significantly enhanced. Interferon-gamma (IFNgamma) was extensively expressed in the non-tolerant T cells from 24 to 48 h after the stimulation with donor alloantigens. In contrast, the expression of IFNgamma was observed in the tolerant T cells from 72 h after the stimulation. Also, the tolerant T cells showed the expression of interleukin-10 (IL-10) and transforming growth factor-beta 1 (TGF-beta1) from 72 h after the stimulation whereas the non-tolerant T cells did not. These data suggest that CY, when administered 2 days after the p.v. injection, induces persistent tolerance by inhibiting T helper type 1 (Th1) activity in the early period but not the Th1 activity in the later periods.

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