Research
Vpu serine 52 dependent counteraction of tetherin is required for HIV-1 replication in macrophages, but not in ex vivo human lymphoid tissue
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* Corresponding author: Michael Schindler michael.schindler@hpi.uni-hamburg.de
1 Heinrich-Pette-Institute for Experimental Virology and Immunology, Martinistrasse 52, 20251 Hamburg, Germany
2 Institute of Virology, University of Ulm, Albert-Einstein-Allee 11, 89081 Ulm, Germany
3 Current address: Emory University, Atlanta GA 30322, USA
Retrovirology 2010, 7:1 doi:10.1186/1742-4690-7-1
Published: 15 January 2010Additional files
Additional file 1:
Supplementary Figure S1. Assessment of viral release by quantitative WB correlates with p24 ELISA. Correlation of the quantitative WB data shown in Fig. 2 with ELISA results, that were measured before the supernatants were pelleted.
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Additional file 2:
Supplementary Figure S2. HeLa derived P4-CCR5 cells express low levels of tetherin. Western blot analysis of endogenous tetherin expression in primary cells and HeLa-derived P4-CCR5 cells. PBMC were either left untreated or stimulated with 1 μg/ml PHA for 24 hours (PBMC+).
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Additional file 3:
Supplementary Figure S3. Vpu S52A is dispensable for HIV-1 release in primary blood lymphocytes (PBL) and Jurkat T-cells. (A) Replication kinetic of the indicated X4-tropic HIV-1 isolates expressing eGFP via an IRES in PBL cultures. PBLs were infected with 1 ng p24 and analyzed for the amount of GFP+ cells in two or three days intervals. Means +/- SD are calculated from infections of PBLs from two donors with three independent virus stocks. (B) The ability of Vpu S52A to enhance HIV-1 release from Jurkat T-cells is inhibited in a tetherin-dependent manner. 1*10^6 Jurkat cells were electroporated with the different proviral constructs coexpressing GFP and the indicated amount of tetherin expression plasmid as described in the methods section. Two days post electroporation the percentage of GFP+ cells as well as p24 contents of the supernatants were quantified. Presented are means and standard deviations from triplicate electroporations from one representative out of three independent experiments.
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