Es obtained with the LTR-specific primers represent a sum of transcripts originating from the host promoter and of viral transcripts, which also contain LTR sequence at their 3' ends. Therefore, the measured ratio corresponds to the equation a+b/b, where `a' is the number of host-viral chimeric transcripts and `b' the number of viral transcripts (see Supplemental text 2). First, we determined this
Es obtained with the LTR-specific primers represent a sum of transcripts originating from the host promoter and of viral transcripts, which also contain LTR sequence at their 3' ends. Therefore, the measured ratio corresponds to the equation a+b/b, where `a' is the number of host-viral chimeric transcripts and `b' the number of viral transcripts (see Supplemental text 2). First, we determined this
Es obtained with the LTR-specific primers represent a sum of transcripts originating from the host promoter and of viral transcripts, which also contain LTR sequence at their 3' ends. Therefore, the measured ratio corresponds to the equation a+b/b, where `a' is the number of host-viral chimeric transcripts and `b' the number of viral transcripts (see Supplemental text 2). First, we determined this
T the level of gene expression act cooperatively to establish and maintain HIV latency. In conclusion, our study reveals that TI represents a key mechanism that antagonizes proviral gene expression to promote the latency of HIV. Furthermore, it demonstrates several means that could be used to counteract TI for reactivating latent HIV. Future mechanistic studies linking TI and other transcriptional
K of the latent provirus in J-Lat 9.2 cells is TI of the 5'LTR caused by the transcription of the host PP5* gene. This interfering transcription results in the lack of Sp1 and the presence of the elongating RNAPII at the 5'LTR. Moreover, TI can be partially rescued by TNF- stimulation, as documented by the appearance of Sp1 and initiating RNAPII at the 5'LTR. The slight enrichment of the RNAPII in
Effect of Tat on transcription of the provirus. In agreement with the results in Figure 4, expression of Tat alone exerted minimal stimulatory effect on the 5'LTR (Figures 7A-7C, Tat). However, in combination with TNF-, Tat expression activated viral transcription greatly (70-80 of GFP-expressing cells) in all three cell lines (Figures 7A-7C, Tat + TNF-). We further investigated the ability of PM
Ranscription units (Han et al., 2004; Lewinski et al., 2005). In the present work, we extended this knowledge by a detailed analysis of how TI impacts viral transcription in latent cells. Although TI occurs regardless of the orientation of the two promoters, we studied TI phenomenon in two J-Lat cell lines with the same orientation of the host and viral promoters, which allowed us to detect and an
Was administered intravenously to individual animals in four cohorts of monkeys. Virus challenge The origin and preparation of the tissue-culture-derived SHIVAD8-EO stock have been previously described25. One week following MAb infusion, animals were challenged intrarectally with 10 TCID50 of SHIVAD8-EO, and every week thereafter, until a virus infection was established. A pediatric nasal speculum