Abstract
BackgroundStratified human keratinocytes (SHKs) are an essential part of mucosal innate immune response that modulates adaptive immunity to microbes encountered in the environment. The importance of these SHKs in mucosal integrity and development has been well characterized, however their regulatory immunologic role at different mucosal sites, has not. In this study we compared the immune gene expression of SHKs from five different anatomical sites before and after HPV16 transfection using microarray analyses.MethodsIndividual pools of human keratinocytes from foreskin, cervix, vagina, gingiva, and tonsils (HFKs, HCKs, HVKs, HGKs and HTLKs) were prepared. Organotypic (raft) cultures were established for both normal and HPV16 immortalized HFKs, HCKs, HVKs, HGKs and HTLKs lines which stably maintained episomal HPV16 DNA. Microarray analysis was carried out using the HumanHT-12 V4 gene chip (Illumina). Immune gene expression profiles were obtained by global gene chip (GeneSifter) and Ingenuity pathway analysis (IPA) for each individual site, with or without HPV16 transfection.ResultsWe examined site specific innate immune response gene expression in SHKs from all five different anatomical sites before and after HPV16 transfection. We observed marked differences in SHK immune gene repertoires within and between mucosal tracts before HPV 16 infection. In addition, we observed additional changes in SHKs immune gene repertoire patterns when these SHKs were productively transfected with HPV16. Some immune response genes were similarly expressed by SHKs from different sites. However, there was also variable expression of non-immune response genes, such as keratin genes, by the different SHKs.ConclusionsOur results suggest that keratinocytes from different anatomical sites are likely hard wired in their innate immune responses, and that these immune responses are unique depending on the anatomical site from which the SHKs were derived. These observations may help explain why select HPV types predominate at different mucosal sites, cause persistent infection at these sites, and on occasion, lead to HPV induced malignant and benign tumor development.
Highlights
Stratified human keratinocytes (SHKs) are an essential part of mucosal innate immune response that modulates adaptive immunity to microbes encountered in the environment
In this study, we identify the differences and similarities in mRNA gene expression made by stratified keratinocytes obtained from five different anatomical sites, grown in organotypic cultures before and after HPV16 transduction
Our results show that keratinocytes within and outside of a given mucosal tract show different immune gene repertoires that are site specific, and that introducing HPV16 into these cells alters their immune gene expression in a site specific manner
Summary
Stratified human keratinocytes (SHKs) are an essential part of mucosal innate immune response that modulates adaptive immunity to microbes encountered in the environment. The importance of these SHKs in mucosal integrity and development has been well characterized, their regulatory immunologic role at different mucosal sites, has not. Responses of SHKs can cause immune dysregulation (Swamy et al 2010; Nestle et al 2009; Strid et al 2009; Albanesi et al 2005; Tonel and Conrad 2009), they support the maintenance of the mucosal microbiome, via defensin expression, and they preserve mucosal homeostasis (Chung and Dale 2004; Frohm et al 1997). Towards understanding how oral cavity-derived keratinocytes influence adaptive immunity, Wu et al (Wu et al 2011) performed a global gene expression analysis that showed the significant effect of murine oral keratinocytes on adaptive immunity (Wu et al 2011)
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