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Human neutrophils release the Leu-8 lymph node homing receptor during cell
activation
M Berg and SP James
Mucosal Immunity Section, National Institute of Allergy and Infectious
Diseases, National Institutes of Health, Bethesda, MD 20892.
The Leu-8 molecule, the human homologue of the murine MEL-14 peripheral
lymph node homing receptor, is expressed on neutrophils in both species and
may be important in localization of cells to sites of inflammation. Most
circulating human neutrophils express the Leu-8 molecule, and activation of
neutrophils with phorbol myristate acetate causes a rapid decline in Leu-8
membrane fluorescence staining within 15 minutes. Northern blot analysis of
total cellular RNA from neutrophils demonstrated two species of Leu-8
messenger RNA, a major one of 2.4 kb and a minor one of 1.9 kb. Because two
different Leu-8 cDNA clones were obtained from human lymphocytes that were
predicted to encode both transmembrane and phosphatidylinositol
(PI)-anchored forms of the molecule, experiments were conducted to
determine whether Leu-8 is anchored to neutrophils by a PI-anchor. There
was a slight decrease in expression of Leu-8 on neutrophils when they were
treated with PI- specific phospholipase C (PI-PLC). However, Leu-8 was
abundant on neutrophils obtained from a patient with paroxysmal nocturnal
hemoglobinuria. To determine the fate of the Leu-8 molecule during cell
activation, neutrophils were labeled with 125I-anti-Leu-8. During
activation antibody was rapidly lost from the cell surface and was not
internalized, suggesting that Leu-8 is released from the cell membrane
during cell activation. When cell extracts of neutrophils were compared
with extracts of lymphoid cells by sodium dodecyl sulfate- polyacrylamide
gel electrophoresis and immunoblotting, the Leu-8 species expressed on
neutrophils had a significantly higher and more variable relative mobility
(70 to 120 Kd for neutrophils v 70 Kd for Jurkat T cells). In addition,
Leu-8 molecules were detected in the supernatants of activated neutrophils.
These results indicate that human neutrophils express a
high-molecular-weight form of the Leu-8 molecule that has a conventional
transmembrane anchor and is rapidly released from the membrane during
activation. The loss of the Leu-8 membrane glycoprotein during activation
may be a mechanism for rapid alteration of neutrophil adhesion
characteristics.
Volume 76,
Issue 11,
pp. 2381-2388,
12/01/1990
Copyright © 1990 by The American Society of Hematology
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