Baxolele Mhlekude1, Annasara Lenman, Phikolomzi Sidoyi, Jim Joseph, Jochen Kruppa, Charles Bitamazire Businge, Mana Lungisa Mdaka, Frank Konietschke, Andreas Pich, Gisa Gerold, Christine Goffinet, Anwar Suleman Mall. 1. University of Cape Town, Department of Surgery, Groote Schuur Hospital, Old Main Building, 7925, Observatory, South Africa TWINCORE, Centre for Experimental and Clinical Infection Research; a joint venture between the Hannover Medical School and the Helmholtz Centre for Infection Research, Institute of Experimental Virology, 30625, Hannover, Germany Charité - Universitätsmedizin Berlin, Institute of Virology, Charité Campus Mitte, 10117, Berlin, Germany Berlin Institute of Health, Berlin, Germany Umeå University, Department of Clinical Microbiology, Virology & Wallenberg Centre for Molecular Medicine (WCMM), SE-90185, Umeå, Sweden Hannover Medical School, Institute of Toxicology, Core Facility Proteomics, 30629, Hannover, Germany Charité - Universitätsmedizin Berlin, Institut für Biometrie und Klinische Epidemiologie, Charité Campus Mitte, 10117, Berlin, Germany Walter Sisulu University, Faculty of Health Sciences, School of Medicine, Nelson Mandela Drive Campus, 5117, Mthatha, South Africa Walter Sisulu University/Nelson Mandela Academic Hospital, Department of Obstetrics and Gynaecology, Nelson Mandela Drive Campus, 5117, Mthatha, South Africa Walter Sisulu University, Department of Human Biology, Nelson Mandela Drive Campus, 5117, Mthatha, South Africa University of Veterinary Medicine Hannover, Department of Biochemistry, 30559, Hanover, Germany.
Abstract
OBJECTIVE: The cervical mucus plugs are enriched with proteins of known immunological functions. We aimed to characterize the anti-HIV-1 activity of the cervical mucus plugs against a panel of different HIV-1 strains in the contexts of cell-free and cell-associated virus. DESIGN: A cohort of consenting HIV-1-negative and -positive pregnant women in labour was recruited from Mthatha General Hospital in the Eastern Cape province of South Africa, from whom the cervical mucus plugs were collected in 6 M guanidinium chloride with protease inhibitors and transported to our laboratories at -80°C. METHODS: Samples were centrifuged to remove insoluble material and dialysed before freeze-drying and subjecting them to the cell viability assays. The antiviral activities of the samples were studied using luminometric reporter assays and flow cytometry. Time-of-addition and BlaM-Vpr virus-cell fusion assays were used to pin-point the antiviral mechanisms of the cervical mucus plugs, before proteomic profiling using liquid chromatography-tandem mass spectrometry. RESULTS: The proteinaceous fraction of the cervical mucus plugs exhibited anti-HIV-1 activity with inter-individual variations and some degree of specificity among different HIV-1 strains. Cell-associated HIV-1 was less susceptible to inhibition by the potent samples when compared to the cell-free HIV-1. The samples with high antiviral potency exhibited a distinct proteomic profile when compared to the less potent samples. CONCLUSION: The crude cervical mucus plugs exhibit anti-HIV-1 activity, which is defined by a specific proteomic profile.
OBJECTIVE: The cervical mucus plugs are enriched with proteins of known immunological functions. We aimed to characterize the anti-HIV-1 activity of the cervical mucus plugs against a panel of different HIV-1 strains in the contexts of cell-free and cell-associated virus. DESIGN: A cohort of consenting HIV-1-negative and -positive pregnant women in labour was recruited from Mthatha General Hospital in the Eastern Cape province of South Africa, from whom the cervical mucus plugs were collected in 6 M guanidinium chloride with protease inhibitors and transported to our laboratories at -80°C. METHODS: Samples were centrifuged to remove insoluble material and dialysed before freeze-drying and subjecting them to the cell viability assays. The antiviral activities of the samples were studied using luminometric reporter assays and flow cytometry. Time-of-addition and BlaM-Vpr virus-cell fusion assays were used to pin-point the antiviral mechanisms of the cervical mucus plugs, before proteomic profiling using liquid chromatography-tandem mass spectrometry. RESULTS: The proteinaceous fraction of the cervical mucus plugs exhibited anti-HIV-1 activity with inter-individual variations and some degree of specificity among different HIV-1 strains. Cell-associated HIV-1 was less susceptible to inhibition by the potent samples when compared to the cell-free HIV-1. The samples with high antiviral potency exhibited a distinct proteomic profile when compared to the less potent samples. CONCLUSION: The crude cervical mucus plugs exhibit anti-HIV-1 activity, which is defined by a specific proteomic profile.