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Leukocyte Complement: Assembly of the Membrane Attack Complex of Complement by Human Peripheral Blood Leukocytes in the Presence and Absence of Serum^1,2,

From the Department of Molecular Immunology, Scripps Clinic and Research Foundation, La Jolla, California 92037

Abstract

The specific neoantigenic determinants (neoAg) that are indicative of the assembled C5b-9 C complex are generated on the surface of peripheral blood leukocytes (PBL) during collection and processing of blood. Formation of neoAg on PBL could be prevented by collecting blood directly into 20 mM EDTA and, could be induced in vitro by adding autologous serum to isolated PBL that lacked neoAg. When neoAg was induced by the addition of serum containing ¹²⁵I-labeled C8, the C8 was incorporated into a 23S complex which could be eluted from PBL.

A mechanism for neoAg formation on PBL independent of exogenous serum factors was detected when PBL were placed into culture in serum-free medium. Results with metabolic inhibitors and ¹⁴C-leucine suggest that PBL can synthesize C5 and assemble the C5b-9 complex. The possible relevance of these findings to the understanding of mechanisms of cell-mediated cytotoxicity is discussed.

Footnotes

¹ This is Publication Number 1401. This investigation was supported by United States Public Health Service Grants AI 07007 and HL 16411 from the National Institutes of Health.

² A preliminary report of these studies has appeared previously: Sundsmo, J. S., W. P. Kolb, and H. J. Müller-Eberhard. 1977. In Regulatory Mechanisms in Lymphocyte Activation. Edited by D. O. Lucas, Academic Press, New York. P. 359.

³ Dr. Sundsmo is supported by United States Public Health Service Training Grant HL 07195.

⁴ Dr. Curd is a Fellow of the Helen Hay Whitney Foundation.

⁵ Dr. Kolb is the recipient of Research Career Development Award 5 K04 CA 70841 from the United States Public Health Service.

⁶ Dr. Müller-Eberhard is the Cecil H. and Ida M. Green Investigator in Medical Research, Scripps Clinic and Research Foundation.