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A Neutrophil Chemotactic Factor Derived from C'5 upon Interaction of Guinea Pig Serum with Endotoxin¹

From the Immunology Section of the Laboratory of Microbiology, National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland 20014, and the Department of Microbiology of the Johns Hopkins University School of Medicine, Baltimore, Maryland 21215

Abstract

Upon activation of the complement system in guinea pig serum by endotoxin, a factor chemotactic for polymorphonuclear leukocytes is generated. This chemotactic factor has an approximate molecular weight (M.W.) of 15,000 and is independent of three minor chemotactic factors (M.W. ;150,000, 68,000 and <4000) present in normal, heat-inactivated or EDTA-treated guinea pig serum. Purified, radiolabeled guinea pig C'3 and C'5 were used to determine whether the 15,000 M.W. chemotactic factor generated in serum was derived from either of these components. Endotoxin was incubated with guinea pig serum to which had been added either ¹²⁵I C'3 or ¹²⁵I C'5. Gel filtration elution profiles of the reaction mixtures revealed that both C'3 and C'5 were cleaved in endotoxin-treated serum. Moreover, the 15,000 M.W. chemotactic factor was identified with a cleavage product of C'5 but not of C'3. In addition, rabbit anti-guinea pig C'5, but not anti-C'3, significantly inhibited the activity of the 15,000 M. W. chemotactic factor. We conclude that the majority of chemotactic activity found in whole guinea pig serum after interaction with endotoxin is a product cleaved from C'5.

Footnotes

Supported in part by National Science Foundation Grant GB-2597, USPHS Grant AI-02566-09, Contract ONR 248(60) with the Office of Naval Research and USPHS Training Grant No. 5TI-AI 282-03.

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