Recombinant expression of bovine LFA-1 and characterization of its role as a receptor for Mannheimia haemolytica leukotoxin

Thamotharampillai Dileepan, Praveen Thumbikat, Bruce Walcheck, Mathur S. Kannan, Samuel K. Maheswaran

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

Mannheimia (Pasteurella) haemolytica leukotoxin (LktA) is the primary virulence factor contributing to the pathogenesis of lung injury in bovine pneumonic mannheimiosis (BPM), a disease which causes major economic loss to the US cattle industry annually. Recent work from our laboratory using an antibody-based approach has shown that LktA binds to bovine LFA-1 in target cells. While this study suggests that LFA-1 might be a specific receptor, it remains to be conclusively shown that LFA-1 is sufficient to induce susceptibility to LktA. It was of interest to determine if functionally active bovine LFA-1 could be reconstituted on a LFA-1 negative cell line and reconstitute susceptibility to LktA. Here, we report the successful recombinant expression of bovine LFA-1 on the cell surface of the human erythroleukemic K562 cell line. The BoLFA-1 transductant expresses bovine CD18 and CD11a as a heterodimer. We found that LktA binds to both the CD18 and CD11a subunits of BoLFA-1 cells. Exposure of BoLFA-1 cells to LktA, induced tyrosine phosphorylation of the CD18 tail, elevation of intracellular calcium, and cytolysis. This is the first report on recombinant expression of functionally active bovine LFA-1 by transduction into an LktA-non-susceptible human cell line.

Original languageEnglish (US)
Pages (from-to)249-257
Number of pages9
JournalMicrobial Pathogenesis
Volume38
Issue number5-6
DOIs
StatePublished - May 2005

Bibliographical note

Funding Information:
This work was supported by USDA-NRI competitive grant # 35204-9230, and Minnesota Agricultural Experimental Station grant.

Keywords

  • Bovine LFA-1
  • Leukotoxin
  • Mannhemia haemolytica
  • Transductant

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