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Long-lived memory T lymphocyte responses after hantavirus infection.

Van Epps HL, Terajima M, Mustonen J, Arstila TP, Corey EA, Vaheri A, Ennis FA - J. Exp. Med. (2002)

Bottom Line: Six novel CD8(+) CTL epitopes were defined on the N protein and were found to be restricted by various HLA alleles including A2, A28, B7, and B8.Interferon (IFN)-gamma ELISPOT analysis showed that memory CTL specific for these epitopes were present at high frequency in PUUV-immune individuals many years after acute infection in the absence of detectable viral RNA.The frequencies of PUUV-specific CTL were comparable to or exceeded those found in other viral systems including influenza, EBV and HIV, in which CTL responses may be boosted by periodic reinfection or virus persistence.

View Article: PubMed Central - PubMed

Affiliation: Center for Infectious Disease and Vaccine Research, University of Massachusetts Medical School, Worcester, MA 01655, USA.

ABSTRACT
Puumala virus (PUUV) is a hantavirus that causes hemorrhagic fever with renal syndrome (HFRS), which is an important public health problem in large parts of Europe. We examined the memory cytolytic T lymphocyte (CTL) responses in 13 Finnish individuals who had HFRS between 1984 and 1995. In seven of these donors, we detected virus-specific CTL responses against the PUUV nucleocapsid (N) protein after in vitro stimulation with PUUV. Six novel CD8(+) CTL epitopes were defined on the N protein and were found to be restricted by various HLA alleles including A2, A28, B7, and B8. This is the first demonstration of PUUV-specific CTL responses in humans, and the first identification of CTL epitopes on PUUV. In addition, this study provides one of the few characterizations of a human antiviral memory T cell response, without the complicating issues of virus persistence or reinfection. Interferon (IFN)-gamma ELISPOT analysis showed that memory CTL specific for these epitopes were present at high frequency in PUUV-immune individuals many years after acute infection in the absence of detectable viral RNA. The frequencies of PUUV-specific CTL were comparable to or exceeded those found in other viral systems including influenza, EBV and HIV, in which CTL responses may be boosted by periodic reinfection or virus persistence.

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Detection of virus-specific CD8+ memory T cells by IFN-γ ELISPOT. PBMCs were stimulated with 10 μg/ml of the indicated peptides in a 16–18 h assay. Input cell numbers ranged from 2–4 × 106 cells per well. Peptide stimulations were performed in triplicate wells. PBMCs stimulated with a B8-restricted epitope from HCV NS3 (NS31402–1411) was included for comparison with PUUV-specific B8-restricted epitopes. PBMCs incubated with media alone was included as negative controls. The data is presented as the number of IFN-γ producing cells/106 PBMCs, with media control values subtracted. The number of spots in the media control wells ranged from 0 to 5. All ELISPOT experiments were performed at least twice and mean values are shown. (A) PUUV-immune individuals who had high precursor frequencies (>100 specific T cells/106 PBMC) of CD8+ T cells specific for one or more CD8+ epitopes on PUUV N. (B) PUUV-immune individuals who had lower precursor frequencies of CD8+ T cells specific for one or more CD8+ epitopes on PUUV N.
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fig3: Detection of virus-specific CD8+ memory T cells by IFN-γ ELISPOT. PBMCs were stimulated with 10 μg/ml of the indicated peptides in a 16–18 h assay. Input cell numbers ranged from 2–4 × 106 cells per well. Peptide stimulations were performed in triplicate wells. PBMCs stimulated with a B8-restricted epitope from HCV NS3 (NS31402–1411) was included for comparison with PUUV-specific B8-restricted epitopes. PBMCs incubated with media alone was included as negative controls. The data is presented as the number of IFN-γ producing cells/106 PBMCs, with media control values subtracted. The number of spots in the media control wells ranged from 0 to 5. All ELISPOT experiments were performed at least twice and mean values are shown. (A) PUUV-immune individuals who had high precursor frequencies (>100 specific T cells/106 PBMC) of CD8+ T cells specific for one or more CD8+ epitopes on PUUV N. (B) PUUV-immune individuals who had lower precursor frequencies of CD8+ T cells specific for one or more CD8+ epitopes on PUUV N.

Mentions: Of the 13 individuals tested, eight had detectable PUUV-specific T cell responses to one or more PUUV N epitope (Fig. 3) . Three individuals had high frequencies of circulating PUUV N-specific memory T cells (donors 1, 11, and 13; Fig. 3 A), with some epitope-specific T cells present at frequencies as high as 300 per 106 PBMCs. The hierarchy of epitope recognition varied from donor to donor, with different epitope-specific populations dominating in different individuals.


Long-lived memory T lymphocyte responses after hantavirus infection.

Van Epps HL, Terajima M, Mustonen J, Arstila TP, Corey EA, Vaheri A, Ennis FA - J. Exp. Med. (2002)

Detection of virus-specific CD8+ memory T cells by IFN-γ ELISPOT. PBMCs were stimulated with 10 μg/ml of the indicated peptides in a 16–18 h assay. Input cell numbers ranged from 2–4 × 106 cells per well. Peptide stimulations were performed in triplicate wells. PBMCs stimulated with a B8-restricted epitope from HCV NS3 (NS31402–1411) was included for comparison with PUUV-specific B8-restricted epitopes. PBMCs incubated with media alone was included as negative controls. The data is presented as the number of IFN-γ producing cells/106 PBMCs, with media control values subtracted. The number of spots in the media control wells ranged from 0 to 5. All ELISPOT experiments were performed at least twice and mean values are shown. (A) PUUV-immune individuals who had high precursor frequencies (>100 specific T cells/106 PBMC) of CD8+ T cells specific for one or more CD8+ epitopes on PUUV N. (B) PUUV-immune individuals who had lower precursor frequencies of CD8+ T cells specific for one or more CD8+ epitopes on PUUV N.
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Related In: Results  -  Collection

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fig3: Detection of virus-specific CD8+ memory T cells by IFN-γ ELISPOT. PBMCs were stimulated with 10 μg/ml of the indicated peptides in a 16–18 h assay. Input cell numbers ranged from 2–4 × 106 cells per well. Peptide stimulations were performed in triplicate wells. PBMCs stimulated with a B8-restricted epitope from HCV NS3 (NS31402–1411) was included for comparison with PUUV-specific B8-restricted epitopes. PBMCs incubated with media alone was included as negative controls. The data is presented as the number of IFN-γ producing cells/106 PBMCs, with media control values subtracted. The number of spots in the media control wells ranged from 0 to 5. All ELISPOT experiments were performed at least twice and mean values are shown. (A) PUUV-immune individuals who had high precursor frequencies (>100 specific T cells/106 PBMC) of CD8+ T cells specific for one or more CD8+ epitopes on PUUV N. (B) PUUV-immune individuals who had lower precursor frequencies of CD8+ T cells specific for one or more CD8+ epitopes on PUUV N.
Mentions: Of the 13 individuals tested, eight had detectable PUUV-specific T cell responses to one or more PUUV N epitope (Fig. 3) . Three individuals had high frequencies of circulating PUUV N-specific memory T cells (donors 1, 11, and 13; Fig. 3 A), with some epitope-specific T cells present at frequencies as high as 300 per 106 PBMCs. The hierarchy of epitope recognition varied from donor to donor, with different epitope-specific populations dominating in different individuals.

Bottom Line: Six novel CD8(+) CTL epitopes were defined on the N protein and were found to be restricted by various HLA alleles including A2, A28, B7, and B8.Interferon (IFN)-gamma ELISPOT analysis showed that memory CTL specific for these epitopes were present at high frequency in PUUV-immune individuals many years after acute infection in the absence of detectable viral RNA.The frequencies of PUUV-specific CTL were comparable to or exceeded those found in other viral systems including influenza, EBV and HIV, in which CTL responses may be boosted by periodic reinfection or virus persistence.

View Article: PubMed Central - PubMed

Affiliation: Center for Infectious Disease and Vaccine Research, University of Massachusetts Medical School, Worcester, MA 01655, USA.

ABSTRACT
Puumala virus (PUUV) is a hantavirus that causes hemorrhagic fever with renal syndrome (HFRS), which is an important public health problem in large parts of Europe. We examined the memory cytolytic T lymphocyte (CTL) responses in 13 Finnish individuals who had HFRS between 1984 and 1995. In seven of these donors, we detected virus-specific CTL responses against the PUUV nucleocapsid (N) protein after in vitro stimulation with PUUV. Six novel CD8(+) CTL epitopes were defined on the N protein and were found to be restricted by various HLA alleles including A2, A28, B7, and B8. This is the first demonstration of PUUV-specific CTL responses in humans, and the first identification of CTL epitopes on PUUV. In addition, this study provides one of the few characterizations of a human antiviral memory T cell response, without the complicating issues of virus persistence or reinfection. Interferon (IFN)-gamma ELISPOT analysis showed that memory CTL specific for these epitopes were present at high frequency in PUUV-immune individuals many years after acute infection in the absence of detectable viral RNA. The frequencies of PUUV-specific CTL were comparable to or exceeded those found in other viral systems including influenza, EBV and HIV, in which CTL responses may be boosted by periodic reinfection or virus persistence.

Show MeSH
Related in: MedlinePlus