Partial characterization of in vivo chemotactic activity: comparison to human C5a

Abstract

Spontaneous eosinophil chemotactic activity (SECA) can mediate the directed movement of human eosinophils and neutrophils. Preliminary characterization of SECA has been carried out. SECA is nondialyzable and heat-stable (56 degrees C, 30 min). Chromatography on Sephadex G-75 demonstrated that SECA had elutional and functional properties similar to C5a (prepared from endotoxin-activated normal sera). Polyacrylamide gel electrophoresis (PAGE) with the use of 15% bisacrylamide gels of lyophilized, chemotactically active column fractions demonstrated a single protein band of identical electrophoretic mobility from either SECA or C5a preparations. Enzymatic hydrolysis with carboxypeptidase B, a known inhibitor of C5a activity, significantly decreased chemotactic activities of C5a and SECA. The addition of purified anti-C5 to either SECA or C5a significantly inhibited chemotactic activity. SECA is naturally occurring chemotactic activity identical to human C5a. Thus C5a may be an important source of in vivo chemotactic activity in various inflammatory disorders.