The leukotoxin of Pasteurella haemolytica binds to beta(2) integrins on bovine leukocytes.
The putative receptor proteins of Pasteurella haemolytica leukotoxin were isolated from bovine polymorphonuclear neutrophil lysate by affinity chromatography on a leukotoxin-specific monoclonal antibody column to which the leukotoxin was pre-bound. SDS-PAGE of the purified proteins showed four bands at 180 kDa, 170 kDa, 150 kDa and 95 kDa, in addition to the expected 102-kDa leukotoxin band and a series of bands with molecular masses lower than 102 kDa representing the disintegrated leukotoxin. N-terminal amino acid sequencing of the 170-kDa band showed homology with human and murine CD11b. The purified proteins reacted specifically with monoclonal antibodies specific for CD11a, CD11b, CD11c (the alpha chains of beta(2) integrins), and CD18 (the beta chain of beta(2) integrins). Pre-incubation of polymorphonuclear neutrophils with a monoclonal antibody specific for CD18 reduced the cytotoxicity of the leukotoxin to the cells. These results indicate that the leukotoxin binds to the beta(2) integrins on bovine leukocytes, very likely via CD18.
Ambagala
T C
TC
Department of Veterinary and Biomedical Sciences, University of Nebraska, Lincoln, NE 68583, USA.
Ambagala
A P
AP
Srikumaran
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S
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000918:Antibody Specificity; D015703:Antigens, CD / Q000737:chemistry; D001427:Bacterial Toxins / Q000378:metabolism; D018821:CD18 Antigens / Q000097:blood* / Q000737:chemistry / Q000276:immunology; D002417:Cattle; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005098:Exotoxins / Q000378:metabolism*; D006801:Humans; D016177:Macrophage-1 Antigen / Q000097:blood / Q000737:chemistry; D016978:Mannheimia haemolytica / Q000472:pathogenicity* / Q000502:physiology*; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000382:microbiology* / Q000502:physiology*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D001427:Bacterial Toxins; D018821:CD18 Antigens; D005098:Exotoxins; D016177:Macrophage-1 Antigen; C018579:leukotoxin