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dc.contributor.supervisor Burgener, Adam (Medical Microbiology) Ball, T. Blake (Medical Microbiology) en_US
dc.contributor.author Romas, Laura
dc.date.accessioned 2015-09-30T18:21:27Z
dc.date.available 2015-09-30T18:21:27Z
dc.date.issued 2014-06-30 en_US
dc.identifier.citation Romas, L. M. et al. A comparative proteomic analysis of the soluble immune factor environment of rectal and oral mucosa. PLoS One 9, e100820, doi:10.1371/journal.pone.0100820 (2014). en_US
dc.identifier.uri http://hdl.handle.net/1993/30854
dc.description.abstract Objective: The rectal mucosa is highly susceptible to HIV infection. Mucosal fluid contains soluble immune proteins that influence HIV infection, and previous studies have shown unique mucosal protein expression in HIV-exposed seronegative (HESN) populations, which may contribute to reduced HIV susceptibility. However, the key correlates of susceptibility at the rectal mucosa have not been well defined, which is a critical knowledge gap for our understanding of HIV pathogenesis. Methods: Rectal lavage from low risk men was screened for HIV-neutralizing activity in a TZM-bl reporter cell line against an R5-tropic HIV virus. Label-free tandem mass spectrometry was used to characterize soluble proteins within rectal lavage samples from a low-risk cohort of men (n=15), and HESN men who have sex with men (MSM; n=25). Protein expression between populations was compared using adjusted t tests (p<0.05), and was interpreted using hierarchical clustering and DAVID biofunctional analysis. Protein expression was further analyzed using survey data on sexual behaviours. Proteins associated with the HESN population were screened for antiviral activity in TZM-bl and PBMC culture against an R5- and X4-tropic virus. Major Results: Rectal mucosal fluid was able to inhibit HIV infection in vitro by 40% (p<0.05). Mass spectrometry identified 30/341 (9%) proteins deferentially expressed (DE) in HESN MSM. DE proteins held functions in immunity (p=6.68x10-6, p=0.001) and epithelial barrier development (p=1.81x10-4; p=0.01); notably, specific antiproteases were elevated in HESN secretions, two of which were screened for antiviral activity. Serpin B4 (+2.52 L2FD; p=1.09x10-5), showed significant inhibition of HIV in TZM-bl (45% BaL, 34% IIIB; p<0.05) and PBMC culture (37% BaL, 49% IIIB; p<0.05); cystatin A (+1.52 L2FD; p=1.40x10-3) showed no inhibitory effects. Serpin B4 expression was not associated with frequency of oral intercourse (p=0.32), partner viral load (r=0.16; p=0.29) or presence of HIV neutralizing IgA in secretions (p=0.52). Conclusions: This thesis reports the use of proteomics to understand HIV-susceptibility at the rectal mucosa, and identified serpin B4 as a novel antiviral immune correlate in a population of HESN MSM. These results may help guide future studies of prevention technologies, such as microbicides or vaccines, which would ultimately help limit the spread of HIV. en_US
dc.publisher PLoS One en_US
dc.rights info:eu-repo/semantics/openAccess
dc.subject HIV en_US
dc.subject Proteomics en_US
dc.subject Mucosal Immunology en_US
dc.subject Men who have sex with men en_US
dc.subject MSM en_US
dc.subject Rectal Mucosa en_US
dc.subject Antiprotease en_US
dc.title Understanding the mucosal fluid proteome in rectal susceptibility to HIV infection en_US
dc.type info:eu-repo/semantics/masterThesis
dc.degree.discipline Medical Microbiology en_US
dc.contributor.examiningcommittee Coombs, Kevin (Medical Microbiology) Ho, Emmanuel (Pharmacy) en_US
dc.degree.level Master of Science (M.Sc.) en_US
dc.description.note February 2016 en_US


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