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J Clin Virol. 2007 Sep;40(1):80-3. Epub 2007 Jul 20.

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

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Multidisciplinary Oncology Centre, Centre Hospitalier Universitaire Vaudois, CHUV, CH-1011 Lausanne, Switzerland.



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.


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).


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).


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.


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.

[Indexed for MEDLINE]

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