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Functional lymphocyte subsets infiltrating renal allografts
Abstracts
177
TRANSITION FROM DONOR TO RECIPIENT ORIGIN OF LYMPH NODE DERIVED LYMPHOCYTES FOLLOWING HEPATIC TRANSPLANTATION. John Fung, Jake Demetris, Adriana Zeevi, Thomas Starzl, Marilyn Marrari, and Rene Duquesnoy; Department of Surgery and Division of Immunopathology, University of Pittsburgh and the Central Blood Bank of Pittsburgh, Pittsburgh, PA The role of"passenger" lymphocytes in the initiation of allograft rejection is not clear; however, several lines of evidence suggest that these cells are potent potentiators of allorecognition. Experimental depletion of these cells from allografts can prolong the survival of an allograft. In addition, these transplanted lymphocytes retain their functional activity to recognize alloantigens of the recipient, evidenced by graft vs. host reactions. We are able to study the interactions of transplanted lymphocytes with those of the recipient following hepatic transplantation at various intervals in humans. The donor liver preparation contains numerous hilar lymph nodes which are transplanted en bloc into the recipient. Approximately 18% of the total liver transplants done in 1984 required allograft hepatectomy with retransplantation for a variety of causes, including unremitting rejection, primary nonfunction, and technical problems. Lymph nodes were removed from the hilum of these samples and sections sent for histologic evaluation. Single cell suspensions of these lymph nodes were then tested for donor/recipient HLA phenotypes by standard lymphocyte microcytotoxicity assays, by direct enumeration using monoclonal anti-HLA staining, and for patterns of anti-recipient and anti-donor reactivity in primary and secondary proliferation assays. Samples were analyzed as early as 3 days posttransplant and up to 1 yr posttransplant. Histologically, within the first 3 weeks, marked enlargement of hilar lymph nodes was noted, primarily with hypertrophy o f T cell dependent areas within the lymph node. In later stages, the size of the lymph nodes returned to normal (or smaller than normal) size with apparent normal architecture. Monoclonal anti-HLA class I antibody preparations directed against donor or recipient antigens were used to enumerate respective lymphocyte populations. A gradual transition from donor to recipient phenotype was noted, with less than 10-15% of cells bearing donor antigens at 4 weeks posttransplant. This was confirmed using standard N I H microcytotoxicity assays demonstrating donor-to-recipient phenotype transition during the posttransplant period. We could not detect donor antigen bearing cells by this method after 4 weeks posttransplant. Finally, the proliferative capacity and allospecificity of lymph node derived cells was tested. Extracted cells were incubated with either irradiated donor or recipient lymphocytes and thymidine uptake determined after 3 and 6 days. A corresponding responsiveness from recipient-specific to donor-specific reactivity was noted, corresonding to donor-to-recipient phenotype transition. This data suggests that alloactivation of recipient lymphocytes can be initiated by "passenger" donor lymphocytes.
FUNCTIONAL CHARACTERIZATION OF INFILTRATING T LYMPHOCYTES IN HUMAN HEPATIC ALLOGRAFTS.John Fung, Adriana Zeevi, Thomas Starzl,Jake Demetris, Shunzaburo Iwatsuki, and Rene Duquesnoy; Department of Surgery and Division of Immunopathology, University of Pittsburgh and Central Blood Bank of Pittsburgh, Pittsburgh, PA We have employed recently developed techniques in T cell cloning to study the nature and function of infiltrating hepatic allograft T cells. Using the rationale that intragraft T cells are activated during cell mediated damage to the allograft, we were able to show that these cells would propagate and remain functionally active in the presence of interleukin-2 (IL-2). In this system, recombinant IL-2,
177
TRANSITION FROM DONOR TO RECIPIENT ORIGIN OF LYMPH NODE DERIVED LYMPHOCYTES FOLLOWING HEPATIC TRANSPLANTATION. John Fung, Jake Demetris, Adriana Zeevi, Thomas Starzl, Marilyn Marrari, and Rene Duquesnoy; Department of Surgery and Division of Immunopathology, University of Pittsburgh and the Central Blood Bank of Pittsburgh, Pittsburgh, PA The role of"passenger" lymphocytes in the initiation of allograft rejection is not clear; however, several lines of evidence suggest that these cells are potent potentiators of allorecognition. Experimental depletion of these cells from allografts can prolong the survival of an allograft. In addition, these transplanted lymphocytes retain their functional activity to recognize alloantigens of the recipient, evidenced by graft vs. host reactions. We are able to study the interactions of transplanted lymphocytes with those of the recipient following hepatic transplantation at various intervals in humans. The donor liver preparation contains numerous hilar lymph nodes which are transplanted en bloc into the recipient. Approximately 18% of the total liver transplants done in 1984 required allograft hepatectomy with retransplantation for a variety of causes, including unremitting rejection, primary nonfunction, and technical problems. Lymph nodes were removed from the hilum of these samples and sections sent for histologic evaluation. Single cell suspensions of these lymph nodes were then tested for donor/recipient HLA phenotypes by standard lymphocyte microcytotoxicity assays, by direct enumeration using monoclonal anti-HLA staining, and for patterns of anti-recipient and anti-donor reactivity in primary and secondary proliferation assays. Samples were analyzed as early as 3 days posttransplant and up to 1 yr posttransplant. Histologically, within the first 3 weeks, marked enlargement of hilar lymph nodes was noted, primarily with hypertrophy o f T cell dependent areas within the lymph node. In later stages, the size of the lymph nodes returned to normal (or smaller than normal) size with apparent normal architecture. Monoclonal anti-HLA class I antibody preparations directed against donor or recipient antigens were used to enumerate respective lymphocyte populations. A gradual transition from donor to recipient phenotype was noted, with less than 10-15% of cells bearing donor antigens at 4 weeks posttransplant. This was confirmed using standard N I H microcytotoxicity assays demonstrating donor-to-recipient phenotype transition during the posttransplant period. We could not detect donor antigen bearing cells by this method after 4 weeks posttransplant. Finally, the proliferative capacity and allospecificity of lymph node derived cells was tested. Extracted cells were incubated with either irradiated donor or recipient lymphocytes and thymidine uptake determined after 3 and 6 days. A corresponding responsiveness from recipient-specific to donor-specific reactivity was noted, corresonding to donor-to-recipient phenotype transition. This data suggests that alloactivation of recipient lymphocytes can be initiated by "passenger" donor lymphocytes.
FUNCTIONAL CHARACTERIZATION OF INFILTRATING T LYMPHOCYTES IN HUMAN HEPATIC ALLOGRAFTS.John Fung, Adriana Zeevi, Thomas Starzl,Jake Demetris, Shunzaburo Iwatsuki, and Rene Duquesnoy; Department of Surgery and Division of Immunopathology, University of Pittsburgh and Central Blood Bank of Pittsburgh, Pittsburgh, PA We have employed recently developed techniques in T cell cloning to study the nature and function of infiltrating hepatic allograft T cells. Using the rationale that intragraft T cells are activated during cell mediated damage to the allograft, we were able to show that these cells would propagate and remain functionally active in the presence of interleukin-2 (IL-2). In this system, recombinant IL-2,