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Updated: Aug 8, 2026
Isolation and Flow Cytometric Analysis of Human Endocervical Gamma Delta T Cells
Published on: February 6, 2017
Quantification of HIV-1-specific T-cell responses at the mucosal cervicovaginal surface
B L Shacklett1, S Cu-Uvin, T J Beadle
1Aaron Diamond AIDS Research Center, The Rockefeller University, New York, NY 10016, USA.
Insights
Detecting HIV-1 specific T cells in cervicovaginal samples is possible using ELISPOT. Women not on antiretroviral therapy showed stronger mucosal immune responses than those on HAART.
Area of Science:
- Immunology
- Virology
- Gynecology
Background:
- Cellular immune responses are crucial for controlling HIV-1 infection.
- Mucosal immunity plays a key role in HIV-1 pathogenesis and transmission.
- Assessing immune responses at mucosal sites is challenging but vital.
Purpose of the Study:
- To characterize HIV-1 specific cellular immune responses at mucosal surfaces.
- To evaluate the utility of the enzyme-linked immuno-spot (ELISPOT) technique for this purpose.
- To compare immune responses in women with and without highly active antiretroviral therapy (HAART).
Main Methods:
- Cervicovaginal mononuclear cells were collected via cytobrush and lavage.
- Interferon-gamma (IFN-gamma) production was measured using ELISPOT assay upon HIV-1 antigen stimulation.
- Responses were compared between HIV-1-positive women on HAART (n=9) and those not on HAART (n=12).
Main Results:
- HIV-1 specific IFN-gamma secreting cells were detected in cervicovaginal samples from 42% of women not on HAART versus 11% of women on HAART.
- Peripheral blood mononuclear cells showed significantly higher HIV-1 specific IFN-gamma responses in women not on HAART compared to those on HAART (P=0.009).
- Mucosal and peripheral T cell recognition of antigens largely overlapped, with some exceptions.
Conclusions:
- HIV-1 specific T cells can be detected in noninvasive gynecological specimens.
- Cervicovaginal HIV-1 specific T cell responses appear less frequent in individuals on HAART.
- These findings highlight potential differences in mucosal immunity modulation by HAART.
Objective:
To characterize HIV-1 specific cellular immune responses at mucosal surfaces using a rapid, sensitive enzyme-linked immuno-spot (ELISPOT) technique.
Design:
Cervicovaginal mononuclear cells obtained from cytobrush and cervicovaginal lavage were assessed for production of interferon-gamma (IFN-gamma) in response to stimulation by HIV-1 antigens. HIV-1 specific responses were compared in a cross-sectional study of two HIV-1-positive patient groups: women not currently on antiretroviral therapy with peripheral CD4 cell counts > 250 x 10(6)/l (n = 12); and women on highly active antiretroviral therapy (HAART) (n = 9).
Methods:
Mononuclear cells from peripheral blood or cervicovaginal specimens were assessed in an ELISPOT assay for responses to HIV-1 antigens expressed by recombinant vaccinia viruses. This assay detects primarily CD8 T cells and shows good correlation with MHC class I tetramer staining of cytotoxic T lymphocytes.
Results:
HIV-1 specific IFN-gamma spot-forming cells were detected in cervicovaginal samples of one out of nine women (11%) on HAART and five out of 12 women (42%) not currently on HAART. In peripheral blood mononuclear cells, HIV-1 specific IFN-gamma spot-forming cells were significantly more numerous in women not currently on HAART than in women on HAART (P = 0.009). In most cases, antigens recognized by mucosal T cells were also recognized by PBMC; however, there were exceptions.
Conclusions:
HIV-1-specific antigen-reactive T cells may be detected in routine, noninvasive gynecological specimens. The results suggest that cervicovaginal HIV-1-specific T cells may be less numerous in individuals on HAART than in those not on HAART, as shown previously for HIV-1-specific cytotoxic T lymphocytes in the peripheral blood.
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