Polyclonal expansion of cervical cytobrush-derived T cells to investigate HIV-specific responses in the female genital tract
Article first published online: 17 AUG 2009
© 2010 The Authors. Journal Compilation © 2010 Blackwell Publishing Ltd
Volume 130, Issue 1, pages 23–33, May 2010
How to Cite
Bere, A., Denny, L., Burgers, W. A. and Passmore, J.-A. S. (2010), Polyclonal expansion of cervical cytobrush-derived T cells to investigate HIV-specific responses in the female genital tract. Immunology, 130: 23–33. doi: 10.1111/j.1365-2567.2009.03172.x
- Issue published online: 6 APR 2010
- Article first published online: 17 AUG 2009
- Received 18 June 2009; revised 23 July 2009; accepted 3 August 2009.
- human immunodeficiency virus;
- T cell
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.