While many virulence factors promoting Streptococcus pyogenes invasive disease have been described, specific streptococcal factors and host properties influencing asymptomatic mucosal carriage remain uncertain. To address the need for a refined model of prolonged S. pyogenes asymptomatic mucosal colonization, we have adapted a preestrogenized murine vaginal colonization model for S. pyogenes. In this model, derivatives of strains HSC5, SF370, JRS4, NZ131, and MEW123 established a reproducible, asymptomatic colonization of the vaginal mucosa over a period of typically 3 to 4 weeks' duration at a relatively high colonization efficiency. Prior treatment with estradiol prolonged streptococcal colonization and was associated with reduced inflammation in the colonized vaginal epithelium as well as a decreased leukocyte presence in vaginal fluid compared to the levels of inflammation and leukocyte presence in non-estradiol-treated control mice. The utility of our model for investigating S. pyogenes factors contributing to mucosal carriage was verified, as a mutant with a mutation in the transcriptional regulator catabolite control protein A (CcpA) demonstrated significant impairment in vaginal colonization. An assessment of in vivo transcriptional activity in the CcpA ؊ strain for several known CcpA-regulated genes identified significantly elevated transcription of lactate oxidase (lctO) correlating with excessive generation of hydrogen peroxide to self-lethal levels. Deletion of lctO did not impair colonization, but deletion of lctO in a CcpA ؊ strain prolonged carriage, exceeding even that of the wild-type strain. Thus, while LctO is not essential for vaginal colonization, its dysregulation is deleterious, highlighting the critical role of CcpA in promoting mucosal colonization. The vaginal colonization model should prove effective for future analyses of S. pyogenes mucosal colonization.
The Gram-positive pathogen Streptococcus pyogenes, or group A streptococcus, is responsible for a wide variety of clinical manifestations ranging from the relatively benign and superficial otitis media, impetigo, and pharyngitis to less common and more invasive conditions, including necrotizing fasciitis and toxic shock syndrome, in addition to the postinfectious complications rheumatic fever and glomerulonephritis (1). The ability to produce such a diversity of infections in so many different tissue compartments is testament to the extensive plasticity of the S. pyogenes transcriptome and an abundance of secreted virulence factors (2). While not considered normal human flora, S. pyogenes can be identified as a colonizer of mucosal surfaces of the oropharynx, rectum, and vaginal mucosa, and a prolonged asymptomatic carriage state can develop (3-7). How S. pyogenes carriage exists on a mucosal surface without inducing disease is poorly understood, but the issue is significant in terms of gaining an improved understanding of host-pathogen interactions and the regulation of mucosal immunity. Furthermore, asymptomatic oropharyngeal carri...