Aberrant capping of membrane proteins on neutrophils from patients with leukocyte adhesion deficiency.
Several functional defects have been found in neutrophils from leukocyte adhesion deficiency (LAD) patients who fail to express the CD11/CD18 leukoadhesins: Mo1, LFA-1, and p150,95. To better understand the functional defects of LAD neutrophils, we have performed capping experiments. Purified normal or LAD neutrophils were labeled with fluorochrome-conjugated concanavalin A (Con A) or F(ab')2 fragments of antiurokinase-type plasminogen activator receptor (uPAR), anti-Fc gamma RIII (CD16), anti-Mo5, and anti-CD14 antibodies. F(ab')2-labeled cells were capped using a second-step F(ab')2 fragment of an antimurine Fab antiserum. Cells were capped for 30 minutes at 37 degrees C, then observed by fluorescence microscopy. LAD neutrophils were found to be deficient in capping, but not clustering of all of the reagents tested to date. The percent of cells exhibiting capping of Con A, Fc gamma RIII, urokinase receptor, CD14, and Mo5 were 52%, 67%, 70%, 25%, and 64% for normal neutrophils but were only 10%, 5%, 2%, 3%, and 1%, respectively, for LAD neutrophils. Capping of this panel of membrane components in LAD or normal neutrophils was not augmented by the addition of either 10(-5) mol/L colchicine or 10(-7) mol/L FMLP. Because capping requires membrane-to-cytosol communication and an intact microfilament linkage, we suggest that leukoadhesins may play a broad role in promoting the redistribution of membrane components including adherence-related receptors such as Fc gamma RIII and the urokinase receptor.
Kindzelskii
A L
AL
Department of Biological Sciences, Wayne State University, Detroit, MI 48202.
Xue
W
W
Todd
R F
RF
Boxer
L A
LA
Petty
H R
HR
D000818:Animals; D002448:Cell Adhesion; D030342:Genetic Diseases, Inborn / Q000097:blood*; D006801:Humans; D016177:Macrophage-1 Antigen / Q000032:analysis / Q000502:physiology*; D008565:Membrane Proteins / Q000502:physiology*; D051379:Mice; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000502:physiology*; D017452:Receptors, IgG / Q000032:analysis / Q000502:physiology*
D016177:Macrophage-1 Antigen; D008565:Membrane Proteins; D017452:Receptors, IgG; D009240:N-Formylmethionine Leucyl-Phenylalanine