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Journal of Histochemistry and Cytochemistry, Vol. 45, 1461-1468, Copyright © 1997 by The Histochemical Society, Inc.

RAPID COMMUNICATION

Receptors for the Chemoattractants C5a and IL-8 Are Clustered on the Surface of Human Neutrophils

Gary D. Graya, Sharon R. Hasslenc, Julia A. Ember, David F. Carneyd, Michael J. Herrona, Stanley L. Erlandsenb, and Robert D. Nelsona
a Department of Dermatology, University of Minnesota School of Medicine, Minneapolis, Minnesota
b Department of Cell Biology and Neuroanatomy, University of Minnesota School of Medicine, Minneapolis, Minnesota
c Genentech, South San Francisco, California
d Scripps Research Institute, La Jolla, California

Correspondence to: Robert D. Nelson, Dept. of Dermatology, Box 98 UMHC, University of Minnesota, Minneapolis, MN 55455.

We have used high-resolution field emission scanning electron microscopy with backscatter electron imaging to detect immunogold-labeled C5a and interleukin-8 (IL-8) receptors on human blood neutrophils. The receptors were labeled with receptor-specific antibodies in combination with secondary antibody conjugated to immunogold. When neutrophils were isolated in a "nonactivated" state, both of these receptor populations were expressed primarily in clusters on nonprojecting domains of the cell membrane. When these cells were double labeled for C5a and IL-8 receptors, intermixing of these receptor species in a common cluster was not found. When neutrophils were isolated in an "activated" state, by mixing the blood with N-formylmethionyl-leucyl-phenylalanine, the cells were seen to be elongated and ruffled at their anterior pole, but the C5a receptors did not disperse or redistribute on the surface of the peptide-activated cells. Analysis of the distribution of human C5a receptors expressed by transfected mouse L-cell fibroblasts showed the C5a receptors to be clustered, but expressed on nonprojecting and projecting domains of the cell surface. These observations provide new information on the topographical expression of leukocyte receptors involved in directing cell migration. (J Histochem Cytochem 45:1461-1467, 1997)

Key Words: neutrophil, mouse L-cells, immunocytochemistry, field emission scanning electron microscopy (FESEM), chemoattractant receptor, C5a receptor (CD88), IL-8 receptor, complement, chemokine


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