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Real time PCR for the assessment of CD8+ T cellular immune response after prophylactic vaccinia vaccination


Trojan, A; Rajeswaran, R; Montemurro, M; Mütsch, Margot; Steffen, Robert (2007). Real time PCR for the assessment of CD8+ T cellular immune response after prophylactic vaccinia vaccination. Journal of Clinical Virology, 40(1):80-83.

Abstract

BACKGROUND: The magnitude of specific CD8+ T cell reactivity responsible for vaccine-induced protection against smallpox infection has not yet been fully elucidated. Among other techniques, RT-PCR for the monitoring of cytokine release in effector T cells against tumor and viral antigens has demonstrated a novel promising method. OBJECTIVE: To determine the functional status of antigen specific CD8+ T cells in healthy participants before and 4 weeks after prophylactic vaccination (Lister strain) against smallpox using quantitative real-time PCR (qRT-PCR). STUDY DESIGN: Changes of interferon-gamma (IFNgamma) mRNA expression levels on short term ex vivo peptide antigen stimulation were measured. The corresponding specific CD8+ T cell reactivity was then displayed as CD8-normalized IFN-gamma levels (IFN-gamma/CD8 ratio). RESULTS: We found a 5-9 fold increase of CD8+ T cell reactivity in three out of four vaccinated individuals. The kinetics and strength determined in responders reveal a virus specific T cell effector repertoire pre-vaccination and a corresponding functional state after immunization comparable also to data obtained from tetramer- and ELISPOT analysis. CONCLUSIONS: Apart from protective vaccinia-specific neutralizing antibodies, the presence of antigen-specific CD8+ T-cells has been demonstrated after vaccinia vaccination. In concordance with others, results from this PCR-based study indicate that this smallpox vaccine induces strong vaccinia virus-specific CD8+ and IFN-gamma producing T cell responses.

Abstract

BACKGROUND: The magnitude of specific CD8+ T cell reactivity responsible for vaccine-induced protection against smallpox infection has not yet been fully elucidated. Among other techniques, RT-PCR for the monitoring of cytokine release in effector T cells against tumor and viral antigens has demonstrated a novel promising method. OBJECTIVE: To determine the functional status of antigen specific CD8+ T cells in healthy participants before and 4 weeks after prophylactic vaccination (Lister strain) against smallpox using quantitative real-time PCR (qRT-PCR). STUDY DESIGN: Changes of interferon-gamma (IFNgamma) mRNA expression levels on short term ex vivo peptide antigen stimulation were measured. The corresponding specific CD8+ T cell reactivity was then displayed as CD8-normalized IFN-gamma levels (IFN-gamma/CD8 ratio). RESULTS: We found a 5-9 fold increase of CD8+ T cell reactivity in three out of four vaccinated individuals. The kinetics and strength determined in responders reveal a virus specific T cell effector repertoire pre-vaccination and a corresponding functional state after immunization comparable also to data obtained from tetramer- and ELISPOT analysis. CONCLUSIONS: Apart from protective vaccinia-specific neutralizing antibodies, the presence of antigen-specific CD8+ T-cells has been demonstrated after vaccinia vaccination. In concordance with others, results from this PCR-based study indicate that this smallpox vaccine induces strong vaccinia virus-specific CD8+ and IFN-gamma producing T cell responses.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > Epidemiology, Biostatistics and Prevention Institute (EBPI)
Dewey Decimal Classification:610 Medicine & health
Language:English
Date:2007
Deposited On:30 Mar 2009 07:27
Last Modified:05 Apr 2016 13:11
Publisher:Elsevier
ISSN:1386-6532
Additional Information:Elsevier full text article
Publisher DOI:https://doi.org/10.1016/j.jcv.2007.04.022
Official URL:http://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6VJV-4P7FHYM-1-3&_cdi=6104&_user=5294990&_orig=search&_coverDate=09%2F30%2F2007&_sk=999599998&view=c&wchp=dGLzVzz-zSkzk&md5=c2b62252c20920768fb2e6b7e857eaaa&ie=/sdarticle.pdf
PubMed ID:17644471

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