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The Journal of Immunology, 1976, 116: 595-599.
Copyright © 1976 by The American Association of Immunologists, Inc.

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A Simple Method for the Determination of Complement Receptor-Bearing Mononuclear Cells1

Jeffrey A. Gelfand, Anthony S. Fauci, Ira Green and Michael M. Frank

From the Laboratory of Clinical Investigation and the Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20014

Abstract

A simple method is described for the identification of mononuclear cells bearing the complement receptor. Gram-negative bacteria are directly fluoresceinated and then incubated in non-immune human serum, which leads to complement fixation via activation of the alernative complement pathway. Complement receptor-bearing mononuclear cells (CRMC) will form easily readable rosettes with these bacteria. The method requires no antibody preparation or purification, uses human serum rather than purified complement components or mouse serum, and gives results that correlate closely (r = 0.9015) with the EAC method of identifying CRMC. Furthermore, these indicator cells cannot bind to mononuclear cells via the E or IgG Fc receptors, eliminating this source of possible error. This method may be used for the identification of CRMC in suspension or frozen tissue sections.

Footnotes

1 A preliminary report of this work was presented and published in abstracted form in Clinical Research, 23:291, April 1975.




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