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Characterization of a novel respiratory syncytial virus-specific human cytotoxic T-lymphocyte epitope - PubMed

Characterization of a novel respiratory syncytial virus-specific human cytotoxic T-lymphocyte epitope

P J Goulder et al. J Virol. 2000 Aug.

Abstract

Respiratory syncytial virus (RSV) infection is a major cause of morbidity in childhood worldwide. The first human RSV-specific cytotoxic T-lymphocyte epitope to be defined is described. This HLA B7-restricted epitope in nucleoprotein (NP) was detectable in four healthy, B7-positive adult subjects using B7-RSV-NP tetrameric complexes to stain CD8(+) T cells.

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Figures

FIG. 1
FIG. 1

(A) Elispot assay using unsorted and sorted PBMC from subject A (HLA type, A2/3 B7/51 Cw5/7). Responses to the 16-mer NP-38 (amino acid sequence, GFYHILNNPKASLLSL). The frequency of IFN-γ sfc per million input cells per well is shown in gray, and percent sfc among CD8+ T cells is shown in black. (B) Optimization of the epitope within the 16-mer NP-38. The optimal epitope is the 9-mer NL9. Effectors were a peptide-specific CTL line generated from subject A, and targets were HLA B7-matched Epstein-Barr virus-transformed B-lymphoblastoid cells from donor 004-TCH (HLA type, A∗0202/29 B7/B44 Cw7/7). (C) Recognition of processed NL9 peptide by RSV-specific effectors. Effectors were RSV NP-specific NL9-B7-restricted effectors (NL9-B7), as described for panel B, and HIV Gag-specific KK9-A3-restricted effectors (KK9-A3) from donor 019-TCH (HLA type, A3/30 B51/60 Cw 15/16). Targets were B-lymphoblastoid cells from subject B (HLA type, A3/− B7/− Cw7/−). Targets were either pulsed with no peptide, with the RSV NL9 peptide, or with the HIV KK9 peptide or infected with RSV NP-vaccinia virus (vac) or with HIV Gag-vaccinia virus. Thus, open circles represent targets pulsed with no peptide and incubated with NL9-B7-specific CTLs, open squares represent targets pulsed with the RSV NL9 peptide and incubated with NL9-B7-specific effectors, etc.

FIG. 2
FIG. 2

Staining of CD8+ T cells with the HLA B7-NL9 (RSV-NP) tetramer. (A) Peptide-specific line used in assays shown in Fig. 1 from subject A (HLA type, A2/3 B7/51 Cw5/7). (B) PBMC from B7-negative subject E (HLA type, A34/68 B51/72 Cw2/16). (C) PBMC from B7-positive subject C (HLA type, A3/26 B7/38 Cw7/8). (D) PBMC from subject D (HLA type, 11/25 B7/18 Cw7/12). (Top right) A2-HIV Gag CTL clones from subject 161j (HLA type, A2/3 B7/60 Cw3/7). (Bottom right) Staining of the same CTL clone as above but using the A2-HIV Gag tetramer.

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