eprintid: 25742
rev_number: 69
userid: 4055
dir: disk0/00/02/57/42
datestamp: 2015-09-28 19:00:07
lastmod: 2016-11-15 14:29:35
status_changed: 2015-09-28 19:00:07
type: thesis_degree
metadata_visibility: show
contact_email: soni.sankapal@gmail.com
item_issues_count: 0
eprint_status: archive
creators_name: Sankapal, Soni Ramachandra
creators_email: sos28@pitt.edu
creators_id: SOS28
title: Studies on the cellular factors and hormones controlling HIV-1 transmission in an organ culture model
ispublished: unpub
divisions: sch_gsph_infectiousdiseasesmicrobiology
full_text_status: public
keywords: Tight junction, cervical and colonic epithelium, HIV-1 transmission, cytokines, menstrual cycle, reproductive hormones
abstract: There is very little information available on how HIV-1 transmits through mucosal epithelial layer since it does not express HIV-1 receptor. Overall goal of the project is to elucidate the role of cellular factors and reproductive hormones on HIV-1 transmission across ectocervical and colonic mucosa in an organ culture model. We hypothesize that upon exposure to HIV-1, a complex signal transduction network is activated in epithelial cells, which leads to compromised barrier function by disrupted tight junctions and expression of immune mediators, which would recruit immune cells towards the epithelial layer for replication of virus. Furthermore, reproductive hormone might have an effect on HIV-1 acquisition risk. To test the hypothesis, we evaluated in context of tissue structure whether HIV-1 induces tight junction disruption in ectocervical and colon epithelial cells, examined the cellular factors, including inflammatory cytokines that are involved in HIV-1 transmission across ectocervical epithelia and evaluated the effect of reproductive hormone on HIV-1 transmission. Our results in aim 1 showed that after exposure to HIV-1, no significant changes in the tight junction/adherens junction protein expression were observed in ectocervical and colon epithelia. However, these tissues were infected after exposure to HIV-1. Our data thus indicate that HIV-1 transverses the ectocervical/colon mucosal epithelia without profoundly disrupting the tight junction/adherens junction between epithelial cell. In aim 2, we found that after HIV-1 exposure, the level of CXCL10 and CXCL11 messages in ectocervical epithelia were upregulated and such induction of cytokines in ectocervical epithelia was dependent on HIV-1 infectivity.  Furthermore, we measured the expression level of cellular factors in HIV-1 exposed ectocervical epithelia by next generation sequencing. Our results indicate that, cellular genes like IL36A, FMO2, CXCL10, MUC1, SAA1 and IL8 were differentially expressed in ectocervical epithelia exposed to HIV-1 compared to controls. These results suggest that exposure to HIV-1 induces cytokine production and other cellular factors in epithelial cells. In aim 3, we investigated the impact of reproductive hormones on the risk of HIV-1 acquisition by analyzing the susceptibility of ectocervical/vaginal tissues to HIV-1 infection and by comparing the epithelial thickness/tight junction protein expression in ectocervical/vaginal tissues at different phases of menstrual cycle. Our results showed no difference in HIV-1 susceptibility, epithelial layer thickness and tight junction/adherence junction protein expression levels in ectocervical/vaginal tissues at different stages of the menstrual cycle. Taken together, our results suggest that risk of HIV-1 infection in the ectocervical/vaginal region does not vary over the course of menstrual cycle. These findings are of public health importance because they expand our understanding on mechanism of atraumatic HIV-1 transmission in mucosal area that may be important for developing effective strategies for preventing HIV-1 transmission.
date: 2015-09-28
date_type: published
pages: 110
institution: University of Pittsburgh
refereed: TRUE
etdcommittee_type: thesis_advisor
etdcommittee_type: committee_cochair
etdcommittee_type: committee_member
etdcommittee_type: committee_member
etdcommittee_name: Chen, Yue
etdcommittee_name: Gupta, Phalguni
etdcommittee_name: Coyne, Carolyn B
etdcommittee_name: Reinhart, Todd A.
etdcommittee_email: cheny@pitt.edu
etdcommittee_email: pgupta1@pitt.edu
etdcommittee_email: coynec2@pitt.edu
etdcommittee_email: reinhar@pitt.edu
etdcommittee_id: CHENY
etdcommittee_id: PGUPTA1
etdcommittee_id: COYNEC2
etdcommittee_id: REINHAR
etd_defense_date: 2015-07-20
etd_approval_date: 2015-09-28
etd_submission_date: 2015-07-22
etd_release_date: 2015-09-28
etd_access_restriction: immediate
etd_patent_pending: FALSE
thesis_type: dissertation
degree: PhD
citation:   Sankapal, Soni Ramachandra  (2015) Studies on the cellular factors and hormones controlling HIV-1 transmission in an organ culture model.  Doctoral Dissertation, University of Pittsburgh.    (Unpublished)  
document_url: http://d-scholarship-dev.library.pitt.edu/25742/1/Soni_Sankapal_etd__august_30_2015.pdf