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An approach of the cytotoxicity of human complement to porcine endothelial cells


An approach of the cytotoxicity of human complement to porcine endothelial cells



Zhonghua Weishengwuxue He Mianyixue Zazhi 17(2): 114-117



Complement activation is central to the rejection of discordant xenografts. In order to assess the respective roles of classical and alternative pathways of complement activation, an in vitro model of hyperacute rejection in the pig-to-human donor recipient combination was designed, using a complement-dependent cytotoxicity test with human serum as the source of xenogenic antibodies and complement. The cytotoxic activity of the sera was evaluated by a colorimetric assay using MTT. Pure human serum lysed 58 +- 5% of pig endothelial cells. Selective inhibition of the classical pathway by using C1q-deficient human sera, only 37+-7% of pig endothelial cells were killed (P lt 0.01 versus normal serum). When the alternative pathway was selectively inhibited by using factor B-deficient human serum which could only lyse 42 +- 10% of porcine endothelial cells(P lt 0.01 versus normal serum). Syngeneic normal pig serum and heat-inactivated serum were not cytotoxic. Mixing serum with deficient classical pathway and serum with deficient alternative pathway restored the cytotoxicity to normal levels. Similarly, the cytotoxic activity of deficient serum supplemented with purified C1q of factor B at physiological concentrations reached that of normal human serum. In this model of in vitro hyperacute rejection, both pathways of complement activation were involved, suggesting that regimens designed to inhibit hyperacute rejection of swine xenografts into humans should take into account the activation of alternative complement pathway.

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