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Long-Term (>1 year) Analyses of Chimerism and Tolerance in Mixed Allogeneic Chimeric Mice Using Normal Mouse Combinations

^a First Department of Pathology,
^b Third Department of Internal Medicine, Kansai Medical University, Osaka, Japan

Key Words. Mixed allogeneic chimeras • Pancreas allografts • Mice • Tolerance

Susumu Ikehara, M.D., Ph.D., First Department of Pathology, Kansai Medical University 10-15 Fumizono-cho, Moriguchi City, Osaka 570-8506, Japan. Telephone: 81-6-6993-9429; Fax: 81-6-6994-8283; e-mail: ikehara{at}takii.kmu.ac.jp

We examined the induction of tolerance using pancreas allografts over the long term (>1 year) in mice for the human application of mixed allogeneic bone marrow transplantation (BMT). T cell-depleted BM cells (BMCs) of C57BL/6 (B6) and C3H/He (C3H) mice were transplanted at various ratios into lethally irradiated B6 mice. The percentages of C3H cells in the chimeric mice gradually decreased, finally declining to only a small percentage, except when the ratio of donor to recipient BMCs was 100:1. However, despite the marked decreases in C3H-type cells, all the pancreas allografts of C3H mice were accepted when more than 1% C3H cells were detected in the peripheral blood. To examine the relationships between percentages of transplanted donor cells and acceptance of pancreas allografts, various percentages of donor and recipient BMCs (5% to 30%) were transplanted. It was found that more than 10% donor cells were necessary for the pancreas allografts to be accepted. In vitro assays for mixed lymphocyte reaction and generation of cytotoxic T-lymphocytes revealed that spleen cells in chimeric mice accepting pancreas allografts are tolerant to both host-type and donor-type major histocompatibility complex (MHC) determinants, but show a vigorous responsiveness to third-party MHC determinants. Since donor-type hemopoietic stem cells (HSCs) were detected in the BM and the liver of the chimeric mice, donor-derived HSCs and donor-derived hematolymphoid cells are responsible for the induction of tolerance. It should be noted that the percentage of donor-type HSCs is higher in the liver (6.2%) than in the BM (0.9%).

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