Immunofluorescent localization of pig complement component 3, regardless of the presence or absence of detectable immunoglobulins, in hyperacutely rejected heart xenografts

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Abstract
Rabbit heart xenografts transplanted into the neck of newborn pigs were all hyperacutely rejected within two hours regardless of the presence or absence of detectable endogenous immunoglobulins (Ig). Cryostat tissue sections were prepared from the rejected rabbit hearts and incubated with sheep polyclonal antibodies against pig complement component 3 (C3), pig IgG and pig IgM. Specific immunoreaction was visualized by fluorescein-conjugated antibodies to sheep IgG. C3 was localized mainly on the surfaces of vascular endothelial as well as myocardial cells, and the localization was not dependent upon the presence of pig immunoglobulins within the same tissue. Both pig IgG and IgM were detected only in the heart xenografts transplanted into suckled pigs, whereas no trace of immunoglobulin was found in those transplanted into circulating antibody-free presuckled pigs. Treatment with cobra venom factor (which inhibits complement activity) prior to transplantation prolonged xenograft survival and completely abolished C3 immunostaining. The results provide new evidence at the histochemical level that the alternative pathway of complement is involved in hyperacute xenograft rejection of the species combination (rabbit to pig) used in this study.