Publikation: HIV-induced apoptosis of activated primary CD4+ T lymphocytes is not mediated by Fas-Fas ligand
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Objective:
To investigate the role of the Fas–Fas ligand (FasL) interaction in HIV-1-
induced apoptosis of primary CD4+ T lymphocytes.
Design:
Activated CD4+ T lymphocytes are the main target of HIV, and T-cell
activation leads to the expression of Fas–FasL and enhances HIV-mediated
apoptosis. Phytohemagglutinin-activated primary CD4+ T cells were infected with
HIV; the process of cell death was examined, and whether the dying and dead cells
were the productively infected cells. The modulation of Fas and FasL expression and
its role in HIV-induced cell death was also investigated.
Methods:
The number of viable and dead cells was determined by trypan blue
exclusion. Apoptosis was quantified using an enzyme-linked immunosorbent assay
measuring the release of cytoplasmic histone-associated DNA fragments. The
percentage of HIV-infected cells was determined by FACS analysis, and viral
production was assessed by a p24 core antigen assay. The following three markers,
HIV-gp-120, annexin-V and 7-AAD, were used to monitor the apoptotic process in
HIV-negative and positive cells. Fas and FasL expression was analyzed at the RNA
level by reverse transcription polymerase chain reaction and at the protein level by
flow cytometry. The contribution of Fas–FasL interactions to apoptosis was
examined by blocking experiments using the antagonist ZB4 anti-Fas antibody.
Results:
HIV-induced apoptosis in activated purified CD4+ T lymphocytes required
infectious virus and was dose-dependent. Apoptosis in HIV-infected cultures was
mostly confined to productively infected cells. The expression of Fas and FasL was
not significantly modulated by infection and blocking Fas–FasL interactions did not
reduce the extent of apoptosis.
Conclusions:
HIV-induced apoptosis of activated CD4+ T cells in vitro is confined to
productively infected cells and is not mediated by a Fas–FasL interaction.
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NORAZ, Nelly, Joël GOZLAN, Jacques CORBEIL, Thomas BRUNNER, Stephen A. SPECTOR, 1997. HIV-induced apoptosis of activated primary CD4+ T lymphocytes is not mediated by Fas-Fas ligand. In: AIDS. 1997, 11(14), pp. 1671-1680. ISSN 0269-9370. Available under: doi: 10.1097/00002030-199714000-00003BibTex
@article{Noraz1997HIVin-14311, year={1997}, doi={10.1097/00002030-199714000-00003}, title={HIV-induced apoptosis of activated primary CD4+ T lymphocytes is not mediated by Fas-Fas ligand}, number={14}, volume={11}, issn={0269-9370}, journal={AIDS}, pages={1671--1680}, author={Noraz, Nelly and Gozlan, Joël and Corbeil, Jacques and Brunner, Thomas and Spector, Stephen A.} }
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