Killing of the S and Re forms of Salmonella minnesota via the classical pathway of complement activation in guinea-pig and human sera.
The S (wildtype) and Re form (heptose-deficient, core-defective mutant) of Salmonella minnesota were killed by treatment with normal guinea-pig serum (GPS). Using C4-deficient GPS and serum containing 0.02 M ethyleneglycol-bis-(beta-aminoethylether)-tetraacetic acid and 0.02 M MgCl2 (EGTA-Mg2+) a reduced killing rate was observed. In normal GPS diluted 1:10 containing 0.02 M EGTA-Mg2+ or in C4-deficient GPS diluted 1:10 no killing occurred, whereas the same serum dilution without EGTA-Mg2+ showed a strong bactericidal effect indicating a dependency upon C4 and Ca2+ ions. Furthermore, in contrast to normal human serum (NHS) no killing occurred in a selective complete C1q-deficient human serum. The bactericidal effect, however, could be restored by addition of highly purified C1q; this is a further indication for a dependency upon the classical pathway of C activation. The C-dependent bactericidal activity was totally abolished when phosphate buffer was used, partially reduced in the presence of veronal-buffered saline (VBS), and not affected by tris-(hydroxymethyl)-aminomethane(Tris) or thioglycollate-buffered system EGTA-Mg2+ alone slightly reduced the growth rate of the bacteria whereas disodium ethylene diaminetetraacetate (EDTA) had a bacteriostatic effect on the S-form. The inhibition of the growth of the Re-form by EDTA was amplified by the addition of serum. Pre-incubation of bacteria with serum for absorption of antibodies did not increase the killing rate of such pre-treated bacteria excluding an antibody-mediated bactericidal reaction. Furthermore, pre-treatment of the bacteria with GPS at 0 degrees reduced the serum sensitivity of both types of bacteria.
D000818:Animals; D001770:Blood Bactericidal Activity / Q000187:drug effects; D002021:Buffers; D003167:Complement Activation; D003172:Complement C1 / Q000172:deficiency; D003181:Complement C4 / Q000172:deficiency; D003171:Complement Pathway, Classical; D004492:Edetic Acid / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D006168:Guinea Pigs; D006801:Humans; D009154:Mutation; D012475:Salmonella / Q000187:drug effects / Q000276:immunology*
D002021:Buffers; D003172:Complement C1; D003181:Complement C4; D004533:Egtazic Acid; D004492:Edetic Acid