Journal article
Transfection of non-susceptible cells with Ovis aries recombinant lymphocyte function-associated antigen 1 renders susceptibility to Mannheimia haemolytica leukotoxin
Veterinary microbiology, Vol.125(1), pp.91-99
2007
Handle:
https://hdl.handle.net/2376/103362
PMID: 17590539
Abstract
Mannheimia haemolytica is an important etiological agent of pneumonia in domestic sheep (DS,
Ovis aries). Leukotoxin (Lkt) produced by this organism is the principal virulence factor responsible for the acute inflammation and lung injury characteristic of
M. haemolytica caused disease. Previously, we have shown that the leukocyte-specific integrins, β
2 integrins, serve as the receptor for Lkt. Although it is certain that CD18, the β subunit of β
2 integrins, mediates Lkt-induced cytolysis of leukocytes, it is not clear whether CD18 of all three β
2 integrins, LFA-1, Mac-1 and CR4, mediates Lkt-induced cytolysis of DS leukocytes. Since polymorphonuclear leukocytes, which express all three β
2 integrins, are the leukocyte subset that is most susceptible to Lkt, we hypothesized that all three β
2 integrins serve as the receptor for Lkt. The objective of this study was to determine whether DS LFA-1 serves as a receptor for
M. haemolytica Lkt. We cloned the cDNA for DS CD11a, the α subunit of LFA-1, and co-transfected it along with the previously cloned cDNA for DS CD18, into a Lkt-non-suceptible cell line. Transfectants stably expressing DS LFA-1 were bound by Lkt. More importantly, Lkt lysed the DS LFA-1 transfectants in a concentration-dependent manner. Pre-incubation of Lkt with a Lkt-neutralizing monoclonal antibody (MAb), or pre-incubation of transfectants with MAbs specific for DS CD11a or CD18, inhibited Lkt-induced cytolysis of the transfectants. Exposure of LFA-1 transfectants to low concentrations of Lkt resulted in elevation of intracellular [Ca
2+]
i. Taken together, these results indicate that DS LFA-1 serves as a receptor for
M. haemolytica Lkt.
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Details
- Title
- Transfection of non-susceptible cells with Ovis aries recombinant lymphocyte function-associated antigen 1 renders susceptibility to Mannheimia haemolytica leukotoxin
- Creators
- Paulraj K Lawrence - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USARohana P Dassanayake - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USADonald P Knowles - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USASubramaniam Srikumaran - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USA
- Publication Details
- Veterinary microbiology, Vol.125(1), pp.91-99
- Academic Unit
- Veterinary Microbiology and Pathology, Department of
- Publisher
- Elsevier B.V
- Identifiers
- 99900546572501842
- Language
- English
- Resource Type
- Journal article