LINKED UNRESPONSIVENESS: EARLY CYTOKINE GENE EXPRESSION PROFILES IN CARDIAC ALLOGRAFTS FOLLOWING PRETREATMENT OF RECIPIENTS WITH BONE MARROW CELLS EXPRESSING DONOR MHC ALLOANTIGEN

Linked unresponsiveness operates to induce specific unresponsiveness to fully mismatched vascularized allografts in recipients pretreated with anti-CD4 antibody and syngeneic bone marrow cells expressing a single donor MHC class I alloantigen. The aim of the study was to evaluate early post transplant cytokine expression in allografts where linked unresponsiveness was required for long term graft survival. CBA (H2k) mice were pretreated with CBK (H2k+Kb) bone marrow cells under the cover of anti-CD4 antibody 28 days before transplantation of a CBK or a C57BL/10 (H2b) cardiac allograft. In both cases graft survival was prolonged (MST=100 days). Intragraft expression for interferon (IFN)-γ, interleukin (IL)-2, IL-4, IL-10, IL-12p40, IL-18, iNOS, transforming growth factor (TGF)-β1 and C-β was determined on day 1.5, 3, 7 and 14 after transplantation. Whereas rejecting allografts displayed a sharp peak in IFN-γ, IL-2, IL-4 and IL-10 expression, non-rejecting allografts were characterized by an initial TGF-β1 and IFN-γ production. An increasing IL-4 expression towards day 14 was a unique feature of linked unresponsiveness. All non-rejecting allografts were characterized by an increasing IL-12p40 production towards day 14. In summary, the early cytokine expression pattern in allografts after bone marrow induced operational tolerance is influenced by the quantity of donor alloantigens expressed on the graft as well as on the bone marrow inoculum.