ANALYSIS OF THE B7 COSTIMULATORY PATHWAY IN ALLOGRAFT REJECTION1

Abstract

Blockade of the B7/CD28 costimulation pathway with the fusion protein, CTLA4-Ig, has been shown to prolong allograft survival in numerous rodent models, suggesting that this pathway is functionally important in the allograft rejection response. This pathway is complex and consists of at least the B7-1, B7-1a, B7-1cyt II, and B7-2 molecules on the antigen-presenting cell and CD28 and CTLA4 molecules on the T cell. The intragraft transcript expression of the B7 molecules and their counterreceptors was defined using reverse transcriptase-polymerase chain reaction in the vascularized mouse cardiac allograft model. In addition, the functional significance of these molecules was investigated both in vitro in the mixed leukocyte response (MLR) and in vivo in the vascularized mouse cardiac allograft model. Intragraft expression of B7-1, B7-1a, B7-1cyt II, B7-2, CD28, and CTLA4 transcripts is up-regulated in allografts when compared with both normal untransplanted hearts and syngeneic transplants at between 5 and 12 days after transplant. Both anti-B7-1 and anti-B7-2 monoclonal antibodies alone inhibited T-cell proliferation in the MLR, however, equivalent maximal inhibition was obtained by a combination of these agents or by CTLA4-Ig. Likewise, in the mouse cardiac allograft model, both anti-B7-1 and anti-B7-2 modestly prolonged graft survival. However, an increased survival was obtained with either a combination of anti-B7-1 and anti-B7-2 or CTLA4-Ig. Blockade of the B7/CD28 pathway in the MLR using T cells from CD28 knockout mice had no effect on the proliferative response. Likewise, blockade of the B7/CD28 pathway did not effect the rate of rejection of cardiac allografts by CD28 knockout recipients. These data suggest that both B7-1 and B7-2 have an important role in allograft rejection in the mouse vascularized cardiac allograft model.