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

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

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Cervix Uteri / metabolism*
  • Cervix Uteri / virology
  • Cluster Analysis
  • Cornified Envelope Proline-Rich Proteins / metabolism
  • Cystatin B / metabolism
  • Early Diagnosis
  • Elafin / metabolism
  • Female
  • HIV Infections / metabolism
  • HIV Infections / transmission*
  • High-Throughput Screening Assays
  • Humans
  • Kallikreins / metabolism
  • Longitudinal Studies
  • Male
  • Microfilament Proteins / metabolism
  • Nuclear Proteins / metabolism
  • Proteomics / methods*
  • Sexual Partners
  • Sexually Transmitted Diseases / metabolism*
  • Tandem Mass Spectrometry
  • Vagina / metabolism*
  • Vagina / virology
  • Young Adult

Substances

  • CSTB protein, human
  • Cornified Envelope Proline-Rich Proteins
  • Elafin
  • Microfilament Proteins
  • Nuclear Proteins
  • PI3 protein, human
  • SPRR3 protein, human
  • CAPG protein, human
  • Cystatin B
  • KLK10 protein, human
  • Kallikreins