Streptococcal Pyrogenic Exotoxin B-Induced Apoptosis in A549 Cells Is Mediated through αvβ₃ Integrin and Fas

• Published in: Infection and Immunity 2008-04, Vol.76 (4), p.1349-1357
• Main Authors: Tsai, Wan-Hua, Chang, Chia-Wen, Lin, Yee-Shin, Chuang, Woei-Jer, Wu, Jiunn-Jong, Liu, Ching-Chuan, Tsai, Pei-Jane, Lin, Ming T
• Format: Article
• Language: English
• Subjects: Bacteriology; Biological and medical sciences; Cellular Microbiology: Pathogen-Host Cell Molecular Interactions; Fundamental and applied biological sciences. Psychology; Microbiology; Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains
• ISSN: 0019-9567; 1098-5522
• DOI: 10.1128/IAI.01162-07

Our previous work suggested that streptococcal pyrogenic exotoxin (SPE) B-induced apoptosis is mediated through a receptor-like mechanism. In this study, we have identified αvβ₃ and Fas as the SPE B receptors for this function. The SPE B fragment without the RGD motif and G308S, a SPE B mutant with the RSD motif, induced less apoptosis than did native SPE B, suggesting that the RGD motif is critical for SPE B-induced apoptosis. Fluorescein isothiocyanate-SPE B binding assays and immunoprecipitation analysis showed that SPE B specifically interacted with αvβ₃. Anti-αvβ₃ antibody partially inhibited SPE B-induced apoptosis but had no effect on G308S-induced apoptosis. In addition, Fas binding to SPE B was verified in an affinity column and an immunoprecipitation analysis. Anti-Fas antibody inhibited SPE B- and G308S-induced apoptosis in a dose-dependent manner, suggesting that Fas-mediated SPE B-induced apoptosis also occurs RGD independently. Both anti-αvβ₃ and anti-Fas antibodies synergistically inhibited SPE B-induced apoptosis. The apoptotic cascades were activated by SPE B and G308S, with a little delay by the latter. After SPE B binding, the cell surface level of αvβ₃, but not of Fas, was decreased. The decreased αvβ₃ level was restored by treatment with the proteasome inhibitor MG132, suggesting a SPE B-mediated endocytosis of integrin αvβ₃ via the ubiquitin-proteasome system. Taken together, our results demonstrate that SPE B-induced apoptosis is mediated through αvβ₃ integrin and Fas in a synergistic manner.