Induction of neutrophil homotypic adhesion via sialophorin (CD43), a surface sialoglycoprotein restricted to haemopoietic cells.
CD43 is a cell-surface sialoglycoprotein which is selectively expressed on lympho-haemopoietic cells. We studied the effects of three CD43 antibodies (6E5, 6F5 and 10G7) on human neutrophils and found that all three monoclonal antibodies (mAb) induced significant homotypic adhesion involving more than 50% of cells. Monovalent Fab fragments of CD43 mAb had no such effect but became equally effective upon cross-linkage with F(ab')2 sheep anti-mouse immunoglobulin (Ig) antibodies. The homotypic adhesion induced by CD43 antibodies was dependent on divalent cations, energy, temperature and an intact cytoskeleton, but not on de novo protein synthesis. Homotypic adhesion could be inhibited by mAb to CD11b, CD18 and CD54, indicating an involvement of the beta 2 integrin cyto-adhesion pathway. Additionally, oxidative burst formation was observed with intact CD43 mAb. No such effect was seen with monomeric or cross-linked Fab fragments. This, together with the observation that burst formation unlike adhesion induction could be completely abolished with Fc gamma RII, but not with Fc gamma RIII antibody fragments, suggests that in burst induction, heterologous cross-linkage with Fc gamma RII is involved. A Ca2+ increase with CD43 antibodies was not detectable. Adhesion induction was unaffected by H7, chelerythrin, staurosporine or lavendustin A, but was completely ablated by sphingosine and herbimycin A. This suggests an involvement of tyrosine kinases but not of protein kinase C in the signal transduction cascade leading to homotypic adhesion. CD43 mAb-induced burst formation differed from adhesion induction in that it could be additionally inhibited with staurosporine and lavendustin A.
Rosenkranz
A R
AR
Institute of Immunology, University of Vienna, Austria.
Majdic
O
O
Stöckl
J
J
Pickl
W
W
Stockinger
H
H
Knapp
W
W
D000911:Antibodies, Monoclonal / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology*; D018821:CD18 Antigens; D002448:Cell Adhesion / Q000276:immunology; D015815:Cell Adhesion Molecules / Q000276:immunology; D003599:Cytoskeleton / Q000276:immunology; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D016023:Integrins / Q000276:immunology; D018799:Intercellular Adhesion Molecule-1; D051917:Leukosialin; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism; D010084:Oxidation-Reduction; D017452:Receptors, IgG / Q000276:immunology; D016897:Respiratory Burst / Q000276:immunology; D012795:Sialoglycoproteins / Q000276:immunology*; D015398:Signal Transduction / Q000276:immunology
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018821:CD18 Antigens; D015815:Cell Adhesion Molecules; D007140:Immunoglobulin Fab Fragments; D016023:Integrins; D051917:Leukosialin; D017452:Receptors, IgG; C465014:SPN protein, human; D012795:Sialoglycoproteins; D018799:Intercellular Adhesion Molecule-1