Fig. 3.
Fig. 3. Characterization of EBV specificity of CTL lines expanded from 4 patients with CAEBV. / The EBV specificity of the T-cell lines generated from patients with persistent active EBV infection syndrome was determined by testing the cytotoxic activity of CTLs against a panel of LCL lines, EBV peptide-loaded autologous PHA blasts, or vaccinia virus–infected matched fibroblasts. Data from 20:1 E/T ratios are presented. Panels Ai, Bi, Ci, and D show the CTL recognition of autologous, HLA antigen–partially matched and HLA antigen–mismatched LCLs of 4 representative CTL lines. The CTL presented in panel Ai kills preferentially through B7, whereas intermediate cytotoxicity is directed against LCLs matched for A3 or B65. The CTL presented in panel Bi kills preferentially through B38 and B44, since no significant cytotoxic activity is presented against LCLs matched only for A31 or A26. Panel Ci shows that the T-cell line is prevalently B27-restricted, though A2-, A3-, and B35-restricted killing also occurs. CTLs presented in panel D also kill through B27. Once the HLA antigen restriction was established, the peptide epitope specificity was investigated on autologous or matched PHA blasts loaded with the peptides predicted from the literature (panels Aii, Bii, and C).3549 The HLA antigen class I restriction and antigen location for the peptide epitopes used in the CTL assay are as follows: RPPIFIRRL (B7 EBNA 3A, coordinate 379-387), VPAPAGPIV (B7 EBNA 3C, coordinate 502-510), QPRAPIRPI (B7 EBNA 3A, coordinate 881-889), EENLLDFVRF (B44 EBNA 3C, coordinate 281-290), EGGVGWRHW (B44 EBNA 3C, coordinate 163-171), KEHVIQNAF (B44 EBNA 3C, coordinate 335-343), LLDFVRFMGV (A2 EBNA 3C, coordinate 284-293), SVRDRLARL (A2 EBNA 3A, coordinate 596-604), CLGGLLTMV (A2 LMP2a, coordinate 426-434), DTPLIPLTIF (A2 EBNA 2, coordinate 42-51). In panel E, these 4 lines (CTL line A, closed bar; CTL line B, gray; CTL line C, striped bar; CTL line D, dotted bar) were tested against HLA antigen–matched dermal fibroblasts infected with vaccinia virus recombinants expressing the EBNA 3A, B, and C genes or the lytic BZLF1 gene or GFP gene, as control.

Characterization of EBV specificity of CTL lines expanded from 4 patients with CAEBV.

The EBV specificity of the T-cell lines generated from patients with persistent active EBV infection syndrome was determined by testing the cytotoxic activity of CTLs against a panel of LCL lines, EBV peptide-loaded autologous PHA blasts, or vaccinia virus–infected matched fibroblasts. Data from 20:1 E/T ratios are presented. Panels Ai, Bi, Ci, and D show the CTL recognition of autologous, HLA antigen–partially matched and HLA antigen–mismatched LCLs of 4 representative CTL lines. The CTL presented in panel Ai kills preferentially through B7, whereas intermediate cytotoxicity is directed against LCLs matched for A3 or B65. The CTL presented in panel Bi kills preferentially through B38 and B44, since no significant cytotoxic activity is presented against LCLs matched only for A31 or A26. Panel Ci shows that the T-cell line is prevalently B27-restricted, though A2-, A3-, and B35-restricted killing also occurs. CTLs presented in panel D also kill through B27. Once the HLA antigen restriction was established, the peptide epitope specificity was investigated on autologous or matched PHA blasts loaded with the peptides predicted from the literature (panels Aii, Bii, and C).35 49 The HLA antigen class I restriction and antigen location for the peptide epitopes used in the CTL assay are as follows: RPPIFIRRL (B7 EBNA 3A, coordinate 379-387), VPAPAGPIV (B7 EBNA 3C, coordinate 502-510), QPRAPIRPI (B7 EBNA 3A, coordinate 881-889), EENLLDFVRF (B44 EBNA 3C, coordinate 281-290), EGGVGWRHW (B44 EBNA 3C, coordinate 163-171), KEHVIQNAF (B44 EBNA 3C, coordinate 335-343), LLDFVRFMGV (A2 EBNA 3C, coordinate 284-293), SVRDRLARL (A2 EBNA 3A, coordinate 596-604), CLGGLLTMV (A2 LMP2a, coordinate 426-434), DTPLIPLTIF (A2 EBNA 2, coordinate 42-51). In panel E, these 4 lines (CTL line A, closed bar; CTL line B, gray; CTL line C, striped bar; CTL line D, dotted bar) were tested against HLA antigen–matched dermal fibroblasts infected with vaccinia virus recombinants expressing the EBNA 3A, B, and C genes or the lytic BZLF1 gene or GFP gene, as control.

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