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HLA class I-restricted cytotoxic T-cell epitopes of the respiratory syncytial virus fusion protein - PubMed

HLA class I-restricted cytotoxic T-cell epitopes of the respiratory syncytial virus fusion protein

A H Brandenburg et al. J Virol. 2000 Nov.

Abstract

Virus-specific cytotoxic T lymphocytes (CTL) play a major role in the clearance of respiratory syncytial virus (RSV) infection. We have generated cytotoxic T-cell clones (TCC) from two infants who had just recovered from severe RSV infection. These TCC were functionally characterized and used to identify HLA class I (B57 and C12)-restricted CTL epitopes of RSV.

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Figures

FIG. 1
FIG. 1

Responses of RSV-specific TCC from patients 1 (A) and 2 (B) measured by an IFN-γ ELIspot assay using autologous B-LCL infected with RSV A2 or rVV expressing the RSV F, 1B, or 1C protein. Results are indicated as numbers of spots per well. No responses against the B-LCL infected with the other rVV were found (data not shown).

FIG. 2
FIG. 2

Fine mapping of T-cell epitopes on the RSV F protein. Responsiveness of TCC from patients 1 (A) and 2 (B) to partially overlapping 18-mer peptide amides (n = 94) spanning the F protein was measured by an IFN-γ ELIspot assay. Minimal peptide activation of TCC from patients 1 (C) and 2 (D) was measured in an IFN-γ ELIspot assay using APC pulsed with two additional sets of overlapping 8- to 12-mer peptides. med, medium.

FIG. 3
FIG. 3

Determination of HLA restriction of RSV F protein recognition by specific TCC using a set of autologous and partially HLA class I-matched B-LCL for patients 1 (A) and 2 (B) by an IFN-γ ELIspot assay.

FIG. 4
FIG. 4

Cytotoxic responses (A) and cytokine production (B) of selected TCC with different protein specificities from patients 1 and 2. TCC were stimulated with autologous RSV-infected or control B-LCL and in the case of the F-specific TCC of patient 2, minimal-peptide- or control (measles virus) peptide-loaded B-LCL. Cytotoxic responses were measured in a chromium release assay using an effector-to-target cell ratio of 10. Cytokines in cell-free culture supernatant (IFN-γ, IL-4, IL-5, and IL-10) were measured by enzyme-linked immunosorbent assay. Levels of cytokines in the supernatant of TCC stimulated with control B-LCL were all below the detection limit and are not shown.

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