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Figure 6 | Retrovirology

Figure 6

From: Trans-activation, post-transcriptional maturation, and induction of antibodies to HERV-K (HML-2) envelope transmembrane protein in HIV-1 infection

Figure 6

Evidence of trans-activation and post-transcriptional modification of HERV-K (HML-2) Env TM. A) HERV-K (HML-2) Env mRNA expression was detected using primers designed to bind the SU domain or TM domain (SUprimers or TMprimers respectively) at 0,1 and 2 days post infection. Copies of HERV-K (HML-2) Env detected by TMprimers (plain line) increased by the time of infection compared to the copies detected by SUprimers (dashed line). The graph is a representative experiment of 3 individual independent experiments. Copy number was determined as described previously, and β-Actin was used as reference gene [52]. B) The graph represents cumulative data from 3 independent experiments and shows the ratio of copies of HERV-K (HML-2) Env detected by TMprimers/copies detected by SUprimers from HIV-1LAI infected PBMCs 2 days post-infection. Similar data were obtained using primary isolates (91US_4/R5 tropic and BK132/X4 tropic). C) Nested PCR. The figure shows the amplicons obtained after the second round of PCR (around 500pb). TM mRNA is over-expressed during the peak of viremia at d42. Antiviral treatment induced the transcription of both SU and TM mRNA at a similar level. HIVneg: HIV-1 seronegative low risk donor; OP-1830: HIV-1 seroconverter patient (d0: before infection; d12, d42, d76: after infection without treatment, d104: after treatment); water: the non-template well. (D) Assumed precursor proteins at 75 to 90 kDa and TM subunits at 32 to 38 kDa are visible. Hela-T4 cells are infected by HIV-1LAI in presence (4) or not (3) of 10 μl/ml of tunicamycin. HERK-Env transfected cells (2) were used as positive controls for HERV-K TM expression. Uninfected-untransfected cells were used as control for endogenous HERV-K basal expression (1). (E) Representative images of HERV-K TM extracellular expression on PBMCs. HERM-1811-5 (anti-TM) mouse monoclonal antibody and goat anti-mouse Alexa555 (red) were used to detect extracellular expression of HERV-K (HML-2) TM.

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