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Polyclonal expansion of cervical cytobrush‐derived T cells to investigate HIV‐specific responses in the female genital tract
Author(s) -
Bere Alfred,
Denny Lynette,
Burgers Wendy A.,
Passmore JoAnn S.
Publication year - 2010
Publication title -
immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.297
H-Index - 133
eISSN - 1365-2567
pISSN - 0019-2805
DOI - 10.1111/j.1365-2567.2009.03172.x
Subject(s) - polyclonal antibodies , genital tract , sex organ , human immunodeficiency virus (hiv) , female circumcision , biology , anatomy , medicine , gynecology , immunology , physiology , antibody , genetics
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/10 6 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.