title: Characterization of Mannheimia (Pasteurella) haemolytica leukotoxin interaction with bovine alveolar macrophage beta2 integrins.
abstract: Mannheimia (Pasteurella) haemolytica, the etiologic agent of bovine pneumonic mannheimiosis, produces an exotoxic leukotoxin. The leukotoxin (LktA) is a member of the RTX (repeats in toxin) family of bacterial cytolysins and is distinguished from other toxins by its unique target cell specificity to ruminant leukocytes occurring through binding to a specific receptor. We have demonstrated previously that the beta2 integrin LFA-1 is a receptor for LktA in bovine leukocytes and is involved in leukotoxin-induced biological effects. However the subunits within LFA-1 involved in binding to LktA, and post-binding signaling leading to cellular activation have not been well characterized. The purpose of our study was to pinpoint these precise subunits on bovine alveolar macrophages and to characterize their interaction with LktA. The results in this study indicate that although LktA can efficiently bind to the CD18 subunit of both LFA-1 and Mac-1, post-binding signaling events including elevation of intracellular calcium and CD18 tail phosphorylation are only observed through LFA-1. Furthermore, LktA also binds to the CD11a subunit of LFA-1. LktA binding to CD11a could be inhibited by a small molecule inhibitor of the I(inserted)-domain, the major ligand binding interface on CD11a. I-domain inhibition significantly blunts LktA-induced intracellular calcium elevation and tyrosine phosphorylation of the CD18 tail. Based on our results we suggest that LFA-1 serves as the functional leukotoxin receptor on bovine alveolar macrophages.
authors.affiliation: Department of Veterinary and Biomedical Sciences, University of Minnesota, St. Paul, Minnesota 55108, USA.
authors.forename: Praveen
authors.initials: P
authors.lastname: Thumbikat
authors.forename: Thamotharampillai
authors.initials: T
authors.lastname: Dileepan
authors.forename: Mathur S
authors.initials: MS
authors.lastname: Kannan
authors.forename: Samuel K
authors.initials: SK
authors.lastname: Maheswaran
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial; D001427:Bacterial Toxins / Q000378:metabolism*; D018821:CD18 Antigens / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D005098:Exotoxins / Q000378:metabolism*; D016676:Macrophages, Alveolar / Q000378:metabolism*; D016978:Mannheimia haemolytica / Q000378:metabolism*; D011485:Protein Binding; D021122:Protein Subunits
chemical_list: D000907:Antibodies, Bacterial; D001427:Bacterial Toxins; D018821:CD18 Antigens; D005098:Exotoxins; D021122:Protein Subunits; C018579:leukotoxin; D002118:Calcium