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. 1997 Apr 21;185(8):1423–1433. doi: 10.1084/jem.185.8.1423

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(a) Recognition of SLYNTVATL variants by a SLYNTVATL-specific line from donor 868; E/T 3:1. Lysis of targets pulsed with no peptide subtracted to calculate percent specific lysis. This assay was repeated on three occasions with the same pattern of peptide recognition (data not shown). Bulk-cultured lymphocytes from donor 003 also showed no recognition of the SLHNAVAVL variant (0% specific lysis) compared with recognition of SLYNTVATL (45% specific lysis) (data not shown). (b) Recognition of the HLA-A3–restricted peptide epitope RLRPGGKKK by bulk-cultured CTL from donor 003, and the variant RLRPGGKKC (encoded by proviral DNA within donor 023); E/T ratios as shown. Peptide concentrations, 50 μM. Peptide titration assays were also performed using a peptide-specific line from donor 003: the variant peptide RLRPGGKKC was not recognized at peptide concentrations of below 50 μM, whereas the index peptide was recognized above peptide concentrations of 5 nM (data not shown). (c) Recognition of defined HLA-A*0201–, HLA-B8–, and HLA-B62–restricted CTL epitopes (references 17 and 38) by bulk-cultured CTL from donor 46M (HLA class I -A and -B tissue type: HLA-A1/2 -B8/62) at two timepoints: March, 1995, and December, 1996. Bulk CTL cultured for 15–16 d in each case before assay. E/T ratio, 100:1. Peptide concentrations: 10 μM. A2 Gag, SLYNTVATL. A2 Pol, ILKEPVHGV. B8 17.3, GGKKKYKL. B8 17.8, ELRSLYNTV. B8 24.13, EIYKRWII. B8 24.20, DCKTILKAL. B8 Pol, GPKVKQWPL. B8 Nef 1, WPTVRERM. B8 Nef 8, FLKEKGGL. B62 p24, GLNKIVRMY. B62 Pol, ILKEPVHGVY. B62 Nef, TQGYFPDWQNY. (d) SLYNTVATL-specific killing is not detectable in peptide-specific lines stimulated using SLFNTVATL, but is generated using the index peptide to stimulate PBMC from donor 46M. E/T ratios: SLFNTVATL line, 14:1; SLYNTVATL line, 6:1. Lysis of targets pulsed with no peptide subtracted to calculate percent specific lysis. Assay performed at day 16 of culture; the same result was observed at day 23 of culture (data not shown).