title: Oral and vaginal epithelial cell lines bind and transfer cell-free infectious HIV-1 to permissive cells but are not productively infected.
abstract: The majority of HIV-1 infections worldwide are acquired via mucosal surfaces. However, unlike the vaginal mucosa, the issue of whether the oral mucosa can act as a portal of entry for HIV-1 infection remains controversial. To address potential differences with regard to the fate of HIV-1 after exposure to oral and vaginal epithelium, we utilized two epithelial cell lines representative of buccal (TR146) and pharyngeal (FaDu) sites of the oral cavity and compared them with a cell line derived from vaginal epithelium (A431) in order to determine (i) HIV-1 receptor gene and protein expression, (ii) whether HIV-1 genome integration into epithelial cells occurs, (iii) whether productive viral infection ensues, and (iv) whether infectious virus can be transferred to permissive cells. Using flow cytometry to measure captured virus by HIV-1 gp120 protein detection and western blot to detect HIV-1 p24 gag protein, we demonstrate that buccal, pharyngeal and vaginal epithelial cells capture CXCR4- and CCR5-utilising virus, probably via non-canonical receptors. Both oral and vaginal epithelial cells are able to transfer infectious virus to permissive cells either directly through cell-cell attachment or via transcytosis of HIV-1 across epithelial cells. However, HIV-1 integration, as measured by real-time PCR and presence of early gene mRNA transcripts and de novo protein production were not detected in either epithelial cell type. Importantly, both oral and vaginal epithelial cells were able to support integration and productive infection if HIV-1 entered via the endocytic pathway driven by VSV-G. Our data demonstrate that under normal conditions productive HIV-1 infection of epithelial cells leading to progeny virion production is unlikely, but that epithelial cells can act as mediators of systemic viral dissemination through attachment and transfer of HIV-1 to permissive cells.
authors.affiliation: Department of Oral Immunology, Clinical and Diagnostic Sciences, King's College London Dental Institute, King's College London, London, United Kingdom.
authors.forename: Arinder
authors.initials: A
authors.lastname: Kohli
authors.affiliation: Department of Oral Immunology, Clinical and Diagnostic Sciences, King's College London Dental Institute, King's College London, London, United Kingdom; Department of Obstetrics and Gynecology, Boston University School of Medicine, Boston, Massachusetts, United States of America.
authors.forename: Ayesha
authors.initials: A
authors.lastname: Islam
authors.affiliation: Department of Oral Immunology, Clinical and Diagnostic Sciences, King's College London Dental Institute, King's College London, London, United Kingdom.
authors.forename: David L
authors.initials: DL
authors.lastname: Moyes
authors.affiliation: Department of Oral Immunology, Clinical and Diagnostic Sciences, King's College London Dental Institute, King's College London, London, United Kingdom; Department of Microbiology and Ecology, University of Valencia, Valencia, Spain.
authors.forename: Celia
authors.initials: C
authors.lastname: Murciano
authors.affiliation: Department of Oral Immunology, Clinical and Diagnostic Sciences, King's College London Dental Institute, King's College London, London, United Kingdom.
authors.forename: Chengguo
authors.initials: C
authors.lastname: Shen
authors.affiliation: Department of Oral Immunology, Clinical and Diagnostic Sciences, King's College London Dental Institute, King's College London, London, United Kingdom.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Challacombe
authors.affiliation: Department of Oral Immunology, Clinical and Diagnostic Sciences, King's College London Dental Institute, King's College London, London, United Kingdom.
authors.forename: Julian R
authors.initials: JR
authors.lastname: Naglik
mesh_terms: D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism / Q000821:virology*; D005260:Female; D005786:Gene Expression Regulation; D016679:Genome, Viral / Q000235:genetics; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D009061:Mouth Mucosa / Q000166:cytology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D014158:Transcription, Genetic; D014621:Vagina / Q000166:cytology*; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics; D016662:Virus Integration
chemical_list: D004279:DNA, Viral; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D014764:Viral Proteins