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. 2003 Jan;77(1):291–300. doi: 10.1128/JVI.77.1.291-300.2003

FIG. 3.

FIG. 3.

Blood and mucosal Gag-specific CTL clones recognize the same epitope and display the same TCR β VDJ rearrangement. (A to C) Blood, rectal, and seminal CTL clones, established from volunteer NP002 from the same visit. (A) CTL clones derived from blood (B22), semen (S18), and rectum (R9) recognized the same HIV-1 Gag epitope within aa 269 to 277. (B) The four CTL clones (blood-derived B18 and B22, semen-derived S18, and rectum-derived R9) utilize TCR Vβ22 and present identical rearrangements of their TCR β VDJ segments. The four left lanes contain PCR products from the clonal cDNA amplified with pooled Vβ21 to Vβ25 primers. The four right lanes contain the PCR products from the clonal cDNA amplified with the Vβ22-specific primers. The coding and amino acid sequence of the five clones (including B21) TCR BVDJ region is presented to the right. (C) TCR Vβ22 usage was confirmed by flow cytometry. Expression of Vβ22 fluorescein isothiocyanate-conjugated MAb by the CTL clone B22 (solid peak) is shown in comparison to the isotype control (open peak). (D) PBMC from volunteer NS0909 were probed for the presence of TCR Vβ6 and C20 clone-specific VDJ segment. A positive response was detected with Vβ6 specific primers (lane 2) and C20 clone-specific primers (lane 3). Negative and positive control PCR assays were performed in the absence of Vβ6-specific primers (lane 1) or with the cDNA product from cervical clone C20 (lane 5).