Modulation of bovine neutrophil functions by monoclonal antibodies.
The main objective of the study reported here was to generate a panel of monoclonal antibodies (MAB) to bovine neutrophil surface antigens, and to identify MAB that modulate neutrophil chemotaxis, respiratory burst, and phagocytosis. A further objective was to study MAB reactivity with resting and activated neutrophils, to identify activation antigens and adhesion molecules. A panel of 14 MAB was generated by producing murine hybridomas. Neutrophils incubated with MAB at 4 C for 2 hours were used in chemotaxis, respiratory burst, and phagocytosis assays. Chemotaxis was evaluated in Boyden chambers, using Escherichia coli endotoxin-activated fetal bovine serum as the chemoattractant. Respiratory burst was determined by measuring chemoluminescence of neutrophils incubated with 5-amino-2,3-dihydro-1,4-phthalazinedione, and serum opsonized zymosan. Phagocytosis was determined by flow cytometry, using fluorescein-labeled Staphylococcus aureus. The MAB S7G8, S5F8G10, S7E10, and S5F8B8 enhanced chemotaxis (to > 125% of control). The MAB S7E10 and S8D9 enhanced respiratory burst activity (to > 125% of control), whereas MAB S2G8, S4G10, S8G10, and S5F8B8 caused inhibition (to < 75% of control). The MAB S2G8, S4G10, S8G10, and S5F8G10 enhanced phagocytosis (to > 125% of control). Chemotaxis, respiratory burst, and phagocytosis values of neutrophils not bound with MAB served as controls for comparison. The MAB binding for nonactivated neutrophils (at 4 C) ranged from 9 to 100%, and for activated neutrophils (at 37 C; at 37 C with phorbol myristate acetate) from 90 to 100%. Binding of MAB S4F5, S5F8B8, S6C6, S7E10, S8D9, and S5F8G10 increased when neutrophils had been incubated at 37 C.(ABSTRACT TRUNCATED AT 250 WORDS)
Salgar
S K
SK
Department of Animal Science, University of Maryland, College Park 20742.
Paape
M J
MJ
Alston-Mills
B
B
Peters
R R
RR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D002417:Cattle; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D005260:Female; D005434:Flow Cytometry; D066298:In Vitro Techniques; D008163:Luminescent Measurements; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D010587:Phagocytosis / Q000187:drug effects; D012441:Saccharomyces cerevisiae; D013211:Staphylococcus aureus; D013481:Superoxides / Q000097:blood; D015054:Zymosan / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D013481:Superoxides; D015054:Zymosan