Functional and phenotypic characterization of monoclonal antibodies to bovine L-selectin.
OBJECTIVE
To characterize 2 bovine neutrophil monoclonal antibodies (MAB) as to effects on bovine neutrophil function and their binding antigens on the cell surface of bovine neutrophils.


METHODS
16 healthy, lactating Holstein cattle, 1 calf with leukocyte adhesion deficiency, and 1 age-matched control calf, 2 healthy ewes, and 2 healthy human beings as neutrophil sources.


METHODS
Neutrophil chemotactic and respiratory burst activities and calcium influx, and binding properties of the 2 MAB were determined. Molecular mass of corresponding cell surface antigens also was determined, as was binding of human L-selectin MAB DREG56 to molecules recognized by MAB 11G10 and 2G8 on the surface of bovine neutrophils.


RESULTS
MAB 11G10 and 2G8 inhibited chemotactic activity of bovine neutrophils, up-regulated amplitude of native chemiluminescence, and shortened the time to reach maximal chemiluminescence induced by serum-opsonized zymosan. Crosslinking both MAB with a second antibody induced rapid increase in intracellular free calcium concentration. Binding density of MAB 11G10 and 2G8 to bovine neutrophils treated with trypsin was increased (P < 0.05), compared with that of untreated neutrophils. Neutrophils treated with phosphatidylinositol-specific phospholipase C had decreased (P < 0.05) binding density of MAB 11G10 and 2G8. Binding of the various MAB to neutrophils from calves with bovine leukocyte adhesion deficiency was lower (P < 0.05) than binding to neutrophils from healthy calves. Expression of antigens recognized by the aforementioned MAB on the surface of bovine neutrophils was decreased (P < 0.05) within 10 minutes.


CONCLUSIONS
MAB 11G10 and 2G8 recognized L-selectin molecules on bovine neutrophil membrane. L-Selectin (CD62L) is involved in low-affinity adhesion reactions between leukocytes and L-selectin ligand on postcapillary venular endothelial cells.
Wang
Y
Y
Department of Animal Sciences, University of Maryland, College Park 20742, USA.
Paape
M J
MJ
Leino
L
L
Capuco
A V
AV
Närvä
H
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology / Q000502:physiology*; D000954:Antigens, Surface / Q000032:analysis / Q000502:physiology; D002118:Calcium / Q000032:analysis; D002417:Cattle / Q000276:immunology / Q000502:physiology*; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002634:Chemotaxis, Leukocyte / Q000187:drug effects / Q000502:physiology; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods / Q000662:veterinary; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology / Q000502:physiology; D004731:Endotoxins / Q000494:pharmacology; D005260:Female; D006801:Humans; D015151:Immunoblotting / Q000379:methods / Q000662:veterinary; D019041:L-Selectin / Q000032:analysis / Q000276:immunology* / Q000502:physiology; D007962:Leukocytes / Q000737:chemistry / Q000166:cytology / Q000502:physiology; D008163:Luminescent Measurements; D008297:Male; D009504:Neutrophils / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D010641:Phenotype; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D054801:Phosphoinositide Phospholipase C; D011233:Precipitin Tests / Q000379:methods / Q000662:veterinary; D012756:Sheep; D014357:Trypsin / Q000494:pharmacology; D010738:Type C Phospholipases / Q000494:pharmacology; D015054:Zymosan / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D004731:Endotoxins; D019041:L-Selectin; D015054:Zymosan; D010738:Type C Phospholipases; D054801:Phosphoinositide Phospholipase C; D014357:Trypsin; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D002118:Calcium