Mechanisms of activation of the properdin system. Studies on properdin electrophoretic mobility in agarose activation of the alternative pathway.

The electrophoretic mobility of properdin in agarose with and without EDTA examined in sera from normal subjects and from patients with mesangiocapillary glomerulonephritis, systemic lupus erythematosus, rapidly progressive glomerulonephritis, mesangial IgG-IgA disease, minimal change glomerulonephritis and partial lipodystrophy. In 'EDTA agarose", the properdin arc of normal serum was always cathodal (gamma), whereas in non-EDTA agarose it was always (beta), indicating that agarose activated properdin with its consequent conversion from a cathodal to an anodal form. Using this change in the mobility of properdin to investigate activation of the properdin system, it was found that the lower the C3 concentration of diseased sera, the less able were they to support properdin conversion by non-EDTA agarose. This relationship we interpret as a manifestation of the requirement of an intact C3b feedback pathway for properdin activation. This view was supported experimentally by (i) decreasing ability of non-EDTA agarose to shift properdin mobility in normal serum as it was progressively depleted of components of the alternative pathway by cobra venom factor, C3 nehritic factor or Mg2+, and (ii) the inability of non-EDTA agarose to shift properdin in sera depleted of C3 or factor B, and in serum deficient in C3. The report of other workers that activated properdin causes generation of C3b, coupled with our finding that properdin activation depends on the C3b feedback, indicates that a system exists in which activation of the C3b feedback cycle allows activation of properdin, allowing in turn further amplification of the C3b feedback. That the anodal form of properdin may be a property of activated properdin was shown by our observations that properdin eluted from zymosan was anodal and activated, and that the properdin in the supernatant normal serum incubated with inulin was anodal.