Expression of beta2 integrins (CD11a/CD18) in K562 cells

Scope and method of study. Mannheimia haemolytica is the etiological agent for an economically important pneumonic disease of cattle. A critically important virulence factor for this organism is leukotoxin (LKT), an exotoxin elaborated by M. haemolytica that destroys bovine immune cells sent to protect the host. The overall goal of this project was to characterize the interaction between LKT and the putative host cell receptor bovine CD18. Toward this goal, the strategy was to establish transfected cell lines that stably express bovine CD11a/CD18, human CD11a/CD18, or interspecies hybrids (human CD11a/bovine CD18 and bovine CD11a/human CD18). Inserts were generated by PCR of either bovine or human cDNA with CD11a or CD18 specific primers that flanked the coding sequence and included restriction sites to facilitate directional cloning and subcloning. Following generation of recombinant vectors, transfections were performed in both Vero (adherent) and K562 (non-adherent) cells. Initially, Vero cells were transfected with bovine and human CD18 recombinant vectors alone. (Abstract shortened by UMI.)...