Polyclonal expansion of cervical cytobrush-derived T cells to investigate HIV-specific responses in the female genital tract

Authors

  • Alfred Bere,

    1. Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Observatory, Cape Town
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  • Lynette Denny,

    1. Department Obstetrics and Gynaecology, University of Cape Town and Groote Schuur Hospital, Observatory, Cape Town
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  • Wendy A. Burgers,

    1. Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Observatory, Cape Town
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  • Jo-Ann S. Passmore

    1. Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Observatory, Cape Town
    2. National Health Laboratory Services, Cape Town, South Africa
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Dr J-A.S. Passmore, Institute of Infectious Disease and Molecular Medicine, Division of Medical Virology, Faculty of Health Sciences, University of Cape Town, Anzio Road, Observatory 7925, Cape Town, South Africa. Email: Jo-ann.Passmore@uct.ac.za
Senior author: Jo-Ann S. Passmore

Summary

Human immunodeficiency virus (HIV) -specific T-cell responses are detectable in the female genital tract of HIV-infected women but little is known about their frequency or the factors that influence their detection. We investigated the feasibility of polyclonal in vitro expansion of cervical cytobrush-derived T cells to investigate HIV-specific responses in the female genital tract in HIV-infected women. Cytobrush-derived cervical cells were isolated from 22 HIV-infected women and expanded with anti-CD3 and recombinant interleukin-2. Cervical T-cell lines were investigated for Gag-specific responses by interferon-γ ELISPOT and compared with those detected in matched blood samples. Cervical T-cell lines were established from 16/22 (72·7%) participants. Although the absolute number of CD3± cells recovered after expansion was positively associated with the number of cells isolated ex vivo (P = 0·01; R = 0·62), we observed a significant negative correlation between fold expansion and ex vivo cell number (P = 0·004; R = −0·68). We show that both the magnitude (P = 0·002; R = 0·7) and specific Gag regions targeted by cervical T-cell lines (P < 0·0001; R = 0·5) correlated significantly with those detected in blood. With one exception, cervical interferon-γ T-cell responses to Gag were detected only in HIV-infected women with blood Gag-specific response > 1000 spot-forming units/106 cells. We conclude that cervical Gag-specific T-cell responses in expanded lines are most easily detectable in women who have corresponding high-magnitude Gag-specific T-cell responses in blood.

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