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Mol Cell Proteomics. 2019 Mar;18(3):461-476. doi: 10.1074/mcp.RA118.000757. Epub 2018 Nov 30.

A High-throughput Bead-based Affinity Assay Enables Analysis of Genital Protein Signatures in Women At Risk of HIV Infection.

Author information

1
From the ‡Affinity Proteomics, SciLifeLab, Department of Protein Science, KTH - Royal Institute of Technology, Stockholm, Sweden.
2
§Department of Medicine Solna, Unit of Infectious Diseases, Center for Molecular Medicine, Karolinska University Hospital, Karolinska Institutet, Stockholm, Sweden; frideborg.bradley@ki.se.
3
¶Department of Global Health and Department of Epidemiology Health, University of Washington, Seattle, Washington.
4
‖Department of Medical Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada.
5
**National HIV and Retrovirology Labs, JC Wilt Infectious Disease Centre, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
6
‡‡SciLifeLab, Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden.
7
§§Kenya Medical Research Institute, Nairobi, Kenya.
8
¶¶Department of Obstetrics and Gynecology, University of Nairobi, Nairobi, Kenya.
9
‖‖Department of Medicine, Global Health, and Epidemiology, University of Washington, Seattle, Washington.
10
§Department of Medicine Solna, Unit of Infectious Diseases, Center for Molecular Medicine, Karolinska University Hospital, Karolinska Institutet, Stockholm, Sweden.

Abstract

Women at high risk of HIV infection, including sex workers and those with active genital inflammation, have molecular signatures of immune activation and epithelial barrier remodeling in samples of their genital mucosa. These alterations in the local immunological milieu are likely to impact HIV susceptibility. We here analyze host genital protein signatures in HIV uninfected women, with high frequency of condom use, living in HIV-serodiscordant relationships. Cervicovaginal secretions from women living in HIV-serodiscordant relationships (n = 62) were collected at three time points over 12 months. Women living in HIV-negative seroconcordant relationships (controls, n = 25) were sampled at one time point. All study subjects were examined for demographic parameters associated with susceptibility to HIV infection. The cervicovaginal samples were analyzed using a high-throughput bead-based affinity assay. Proteins involved in epithelial barrier function and inflammation were increased in HIV-serodiscordant women. By combining several methods of analysis, a total of five proteins (CAPG, KLK10, SPRR3, elafin/PI3, CSTB) were consistently associated with this study group. Proteins analyzed using the affinity set-up were further validated by label-free tandem mass spectrometry in a partially overlapping cohort with concordant results. Women living in HIV-serodiscordant relationships thus had elevated levels of proteins involved in epithelial barrier function and inflammation despite low prevalence of sexually transmitted infections and a high frequency of safe sex practices. The identified proteins are important markers to follow during assessment of mucosal HIV susceptibility factors and a high-throughput bead-based affinity set-up could be a suitable method for such evaluation.

KEYWORDS:

Affinity proteomics; HIV; HIV-serodiscordant; Immunology*; Inflammation; Tandem Mass Spectrometry; cervix; female reproductive tract; reproductive immunology; vagina

PMID:
30504243
PMCID:
PMC6398207
[Available on 2020-03-01]
DOI:
10.1074/mcp.RA118.000757

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