Fig. 9: Identification of polypeptides that initiate specific T cell responses in treated and untreated HIV infected individuals. | Nature Communications

Fig. 9: Identification of polypeptides that initiate specific T cell responses in treated and untreated HIV infected individuals.

From: Non-AUG HIV-1 uORF translation elicits specific T cell immune response and regulates viral transcript expression

Fig. 9

(A to D), Peptides potentially encoded by uORFs (see peptides “A02; A03; A06; A07; A08” in panel (A) of Fig. 8) were synthesized and used to screen for T cell responses in PBMCs of HIV-infected individuals. PBMCs from treated (ART) and untreated but aviremic (so called elite controllers, EC) individuals were stimulated with a pool of uORF-derived peptides (POOL) and cultured in the presence of IL-2 and IL-7 cytokines in order to expand peptide-specific memory T cells. On day 7 (not shown) and day 12, T cell responses against the POOL were assessed using IFNγ-Elispot. In addition, on day 12, except for donor EC-3, individual peptides of the pool (A02; A03; A06; A07; A08) were assessed using IFNγ-Elispot. As positive control for T cell expansion and activation, a pool of immunogenic peptides from HIV Env and Gag proteins was also used. A Number of uORF-derived peptides recognized by each HIV-infected individuals. The color code indicates from which uORF the peptides are derived (A02= Blue; A03= Orange; A07&A08= Red). B, C and D left panels, detailed IFNγ-Elispot data from the 3 individuals presenting T cell responses, expressed as spot forming units (SFU) per million PBMCs; right panels pictures of the raw data from the Elispot plates in technical triplicate (n = 3 technical replicates) (B and D), or duplicate (n = 2 technical replicates) (C), POOL (−) and POOL (+): PBMCs expanded with the pool of uORF-derived peptides but restimulated on the day of the Elispot with medium or the POOL, respectively. A02; A03; A06; A07 and A08 name of individual uORF-derived peptides used for re-stimulation. HIV (-) and HIV ( + ): PBMC expanded with the pool of Env- and Gag-derived peptides but re-stimulated for the Elispot assay with medium or the same pool of HIV peptides, respectively. Responses were considered positive when IFNγ production was superior to 20 spots/106 PBMCs and at least twofold higher than background production from cells re-stimulated with medium (dotted lines). SAT: saturated signal, where counts cannot be estimated due to overwhelm IFNγ secretion by activated T cells. Source data are provided as a Source Data file. Error bars correspond to the standard deviation of the mean.

Back to article page