The complement system is involved in acute inflammation but not in the hemorrhage produced by a Bothrops atrox snake venom low molecular mass proteinase

Abstract

Low molecular weight hemorrhagins were purified from crude Bothrops atrox snake venom by gel filtration followed by ionic strength chromatography. The protein fractions obtained, designated HI-1 to HI-8, contained proteins with molecular masses lower than 30 kDa. HI-5, the most representative among of these fractions, exhibited, in vitro, proteolytic and C inactivating properties, as analyzed by proteolysis of a protein substrate, and C system consumptive activities as assayed by reduction of the hemolytic C activity in normal human serum and by cleavage of partially purified component C3. HI-5 hemorrhagin injected i.m. into C-sufficient BALB/c mice induced a local inflammation characterized by edema, accumulation of polymorphonuclear leucocytes (PMN) and hemorrhage. In contrast, when injected into BALB/c mice previously C-depleted, the number of PMN per tissue section, but not hemorrhage, was significantly reduced (129.668±31.341 cells per microscopic field) as compared with the control C-sufficient mice (812.168±111.194 cells per microscopic field). The observations were confirmed by using C5-deficient mice instead of C-depleted mice. The average number of PMN per tissue section in C5-defficient A/J mice was 72.666±19.416 cells per microscopic field. These data indicate that the C system is involved in PMN accumulation, but not in the hemorrhage, at the local induced lesions by low molecular mass B. atrox hemorrhagins. HI-5 apparently is not contaminated with other direct or indirect inflammation mediators, PMN accumulation and hemorrhage, however, an independent phenomenon, could be mediated by the same hemorrhagin proteinase domain.