Transcriptomic analysis of vulvovaginal candidiasis identifies a role for the NLRP3 inflammasome

mBio. 2015 Apr 21;6(2):e00182-15. doi: 10.1128/mBio.00182-15.

Abstract

Treatment of vulvovaginal candidiasis (VVC), caused most frequently by Candida albicans, represents a significant unmet clinical need. C. albicans, as both a commensal and a pathogenic organism, has a complex and poorly understood interaction with the vaginal environment. Understanding the complex nature of this relationship is necessary for the development of desperately needed therapies to treat symptomatic infection. Using transcriptome sequencing (RNA-seq), we characterized the early murine vaginal and fungal transcriptomes of the organism during VVC. Network analysis of host genes that were differentially expressed between infected and naive mice predicted the activation or repression of several signaling pathways that have not been previously associated with VVC, including NLRP3 inflammasome activation. Intravaginal challenge of Nlrp3(-/-) mice with C. albicans demonstrated severely reduced levels of polymorphonuclear leukocytes (PMNs), alarmins, and inflammatory cytokines, including interleukin-1β (IL-1β) (the hallmarks of VVC immunopathogenesis) in vaginal lavage fluid. Intravaginal administration of wild-type (WT) mice with glyburide, a potent inhibitor of the NLRP3 inflammasome, reduced PMN infiltration and IL-1β to levels comparable to those observed in Nlrp3(-/-) mice. Furthermore, RNA-seq analysis of C. albicans genes indicated robust expression of hypha-associated secreted aspartyl proteinases 4, 5, and 6 (SAP4-6), which are known inflammasome activators. Despite colonization similar to that of the WT strain, ΔSAP4-6 triple and ΔSAP5 single mutants induced significantly less PMN influx and IL-1β during intravaginal challenge. Our findings demonstrate a novel role for the inflammasome in the immunopathogenesis of VVC and implicate the hypha-associated SAPs as major C. albicans virulence determinants during vulvovaginal candidiasis.

Importance: Vaginitis, most commonly caused by the fungus Candida albicans, results in significant quality-of-life issues for all women of reproductive age. Recent efforts have suggested that vaginitis results from an immunopathological response governed by host innate immunity, although an explanatory mechanism has remained undefined. Using comprehensive genomic, immunological, and pharmacological approaches, we have elucidated the NLRP3 inflammasome as a crucial molecular mechanism contributing to host immunopathology. We have also demonstrated that C. albicans hypha-associated secreted aspartyl proteinases (SAP4-6 and SAP5, more specifically) contribute to disease immunopathology. Ultimately, this study enhances our understanding of the complex interplay between host and fungus at the vaginal mucosa and provides proof-of-principle evidence for therapeutic targeting of inflammasomes for symptomatic vulvovaginal candidiasis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Candida albicans / genetics
  • Candida albicans / growth & development*
  • Candidiasis, Vulvovaginal / microbiology
  • Candidiasis, Vulvovaginal / pathology*
  • Carrier Proteins / biosynthesis*
  • Carrier Proteins / genetics
  • Disease Models, Animal
  • Female
  • Gene Expression Profiling*
  • Inflammasomes / genetics
  • Inflammasomes / metabolism*
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Molecular Sequence Data
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Neutrophils / immunology
  • Sequence Analysis, DNA
  • Vagina / pathology
  • Virulence Factors / genetics

Substances

  • Carrier Proteins
  • Inflammasomes
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Nlrp3 protein, mouse
  • Virulence Factors

Associated data

  • GEO/GSE67688