SP-D impedes transfer of HIV-1 from multi-layered vaginal epithelium with a distinct gene signature
Surfactant Protein (SP) D is a member of the collectin family of soluble pattern recognition receptors. We have previously shown that a recombinant fragment of SP-D (rhSP-D) inhibits gp120-CD4 interaction and HIV-1 entry in target cells. To potentiate its prophylactic use as a vaginal microbicide, w...
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doaj-2ff94d255fb74f2c8b1db1e1fe097f762020-11-24T23:30:06ZengScience Planet Inc.Canadian Journal of Biotechnology2560-83042017-12-011Special Issue-Supplement26026010.24870/cjb.2017-a244SP-D impedes transfer of HIV-1 from multi-layered vaginal epithelium with a distinct gene signatureHrishikesh Pandit0Kavita Kale1Gargi Thakur2Sushama Rokade3Hidemi Yamamoto4Uday Kishore5Taruna Madan6Raina Fichorova7National Institute for Research in Reproductive Health, Innate Immunity, Mumbai, INDIANational Institute for Research in Reproductive Health, Innate Immunity, Mumbai, INDIANational Institute for Research in Reproductive Health, Innate Immunity, Mumbai, INDIANational Institute for Research in Reproductive Health, Innate Immunity, Mumbai, INDIAHarvard Medical School and Brigham and Women's Hospital, Laboratory of Genital Tract Biology, Boston, MA, USABrunel University, Centre for Infection, Immunity and Disease Mechanisms, London, UNITED KINGDOMNational Institute for Research in Reproductive Health, Innate Immunity, Mumbai, INDIAHarvard Medical School and Brigham and Women's Hospital, Laboratory of Genital Tract Biology, Boston, MA, USASurfactant Protein (SP) D is a member of the collectin family of soluble pattern recognition receptors. We have previously shown that a recombinant fragment of SP-D (rhSP-D) inhibits gp120-CD4 interaction and HIV-1 entry in target cells. To potentiate its prophylactic use as a vaginal microbicide, we determined ex vivo efficacy using organotypic human vaginal-ectocervical epithelia (VEC-100) that closely resemble the native tissues of origin. VEC-100, stratified human vaginal-ectocervical tissues grown on membrane inserts were treated with rhSP-D followed by a challenge with HIV-1 to assess the transfer of HIV-1 through the VEC-100 tissues to PBMCs in the basal submucosal compartment. Treated VEC tissues were subjected to mRNA Illumina microarray analysis. Levels of transcripts encoding for immune mediators, adhesion and tight junction proteins were also evaluated. Effect of rhSP-D on viability, NFκB activation, cytokine secretion and bacterial colonization of cervical vaginal epithelial cells was determined. rhSP-D significantly inhibited HIV-1 transfer from the multi-layered epithelial tissues to the basal PBMCs as compared to HIV-1 alone. Global gene expression profile of HIV-1 challenged VEC-100 tissues revealed differential regulation of genes and pathways majorly involved in inflammation, cell survival and transcription factors. Levels of Guanylate-binding proteins (GBPs) and interferon-inducible proteins were significantly upregulated suggesting an interferon host defense response. rhSP-D showed an inhibition in the levels of GBPs and rescued the cell adhesion molecules such as Claudin 2, 3, 4, 5 and Occludin, known to be down regulated by HIV-1 in primary vaginal cells. Importantly, rhSP-D conditioned VEC tissue supernatants did not enhance susceptibility of target cells to HIV-1. rhSP-D treated vaginal epithelial cells did not show any significant alteration in viability, NFκB activation and levels of immune mediators like IL-1RA, Elafin, SLPI, TGFβ, GRO-α, MIP-3α and RANTES. Bacterial colonization and direct toxicity assays revealed that rhSP-D did not adversely affect growth of vaginal commensals. Blockade of viral movement within the vaginal epithelium, inhibition of detrimental early gene signature and safety profile of rhSP-D suggests that topical formulation comprising rhSP-D may significantly curb the sexual transmission of HIV-1.https://www.canadianjbiotech.com/CAN_J_BIOTECH/Archives/v1/Special Issue-Supplement/cjb.2017-a244.pdf |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Hrishikesh Pandit Kavita Kale Gargi Thakur Sushama Rokade Hidemi Yamamoto Uday Kishore Taruna Madan Raina Fichorova |
spellingShingle |
Hrishikesh Pandit Kavita Kale Gargi Thakur Sushama Rokade Hidemi Yamamoto Uday Kishore Taruna Madan Raina Fichorova SP-D impedes transfer of HIV-1 from multi-layered vaginal epithelium with a distinct gene signature Canadian Journal of Biotechnology |
author_facet |
Hrishikesh Pandit Kavita Kale Gargi Thakur Sushama Rokade Hidemi Yamamoto Uday Kishore Taruna Madan Raina Fichorova |
author_sort |
Hrishikesh Pandit |
title |
SP-D impedes transfer of HIV-1 from multi-layered vaginal epithelium with a distinct gene signature |
title_short |
SP-D impedes transfer of HIV-1 from multi-layered vaginal epithelium with a distinct gene signature |
title_full |
SP-D impedes transfer of HIV-1 from multi-layered vaginal epithelium with a distinct gene signature |
title_fullStr |
SP-D impedes transfer of HIV-1 from multi-layered vaginal epithelium with a distinct gene signature |
title_full_unstemmed |
SP-D impedes transfer of HIV-1 from multi-layered vaginal epithelium with a distinct gene signature |
title_sort |
sp-d impedes transfer of hiv-1 from multi-layered vaginal epithelium with a distinct gene signature |
publisher |
Science Planet Inc. |
series |
Canadian Journal of Biotechnology |
issn |
2560-8304 |
publishDate |
2017-12-01 |
description |
Surfactant Protein (SP) D is a member of the collectin family of soluble pattern recognition receptors. We have previously shown that a recombinant fragment of SP-D (rhSP-D) inhibits gp120-CD4 interaction and HIV-1 entry in target cells. To potentiate its prophylactic use as a vaginal microbicide, we determined ex vivo efficacy using organotypic human vaginal-ectocervical epithelia (VEC-100) that closely resemble the native tissues of origin. VEC-100, stratified human vaginal-ectocervical tissues grown on membrane inserts were treated with rhSP-D followed by a challenge with HIV-1 to assess the transfer of HIV-1 through the VEC-100 tissues to PBMCs in the basal submucosal compartment. Treated VEC tissues were subjected to mRNA Illumina microarray analysis. Levels of transcripts encoding for immune mediators, adhesion and tight junction proteins were also evaluated. Effect of rhSP-D on viability, NFκB activation, cytokine secretion and bacterial colonization of cervical vaginal epithelial cells was determined. rhSP-D significantly inhibited HIV-1 transfer from the multi-layered epithelial tissues to the basal PBMCs as compared to HIV-1 alone. Global gene expression profile of HIV-1 challenged VEC-100 tissues revealed differential regulation of genes and pathways majorly involved in inflammation, cell survival and transcription factors. Levels of Guanylate-binding proteins (GBPs) and interferon-inducible proteins were significantly upregulated suggesting an interferon host defense response. rhSP-D showed an inhibition in the levels of GBPs and rescued the cell adhesion molecules such as Claudin 2, 3, 4, 5 and Occludin, known to be down regulated by HIV-1 in primary vaginal cells. Importantly, rhSP-D conditioned VEC tissue supernatants did not enhance susceptibility of target cells to HIV-1. rhSP-D treated vaginal epithelial cells did not show any significant alteration in viability, NFκB activation and levels of immune mediators like IL-1RA, Elafin, SLPI, TGFβ, GRO-α, MIP-3α and RANTES. Bacterial colonization and direct toxicity assays revealed that rhSP-D did not adversely affect growth of vaginal commensals. Blockade of viral movement within the vaginal epithelium, inhibition of detrimental early gene signature and safety profile of rhSP-D suggests that topical formulation comprising rhSP-D may significantly curb the sexual transmission of HIV-1. |
url |
https://www.canadianjbiotech.com/CAN_J_BIOTECH/Archives/v1/Special Issue-Supplement/cjb.2017-a244.pdf |
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