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Clin. Vaccine Immunol. doi:10.1128/CVI.00392-07
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Monitoring of vaccine-specific IFN- induction in the genital mucosa of mice by real-time RT-PCR

Institute of Microbiology, Centre Hospitalier Universitaire Vaudois and University of Lausanne, CH-1011 Lausanne and NCCR "Molecular Oncology", 1066 Epalinges, Switzerland

^* To whom correspondence should be addressed. Email: dnardell{at}hospvd.ch.

	Abstract

Monitoring of T cell responses in genital mucosa has remained a major challenge, due to the absence of lymphoid aggregates and the low abundance of T cells. Here we have adapted to genital tissue a sensitive real-time RT-PCR (TaqMan) method to measure induction of IFN- mRNA transcription after three hours antigen-specific activation of CD8 T cells. For this purpose, we vaccinated C57BL/6 mice subcutaneously with Human Papillomavirus type 16 L1 Virus-like particles and followed the induction of CD8 T cells specific to the L1_165-173 H-2D^b-restricted epitope. Comparison of the responses induced in peripheral blood mononuclear cells (PBMC) and lymph nodes (LN) using L1-specific IFN- ELISPOT and TaqMan determination of the fold increase of L1-specific IFN- mRNA induction normalized to the content in CD8b mRNA showed a significant correlation, despite the difference in readouts. Most of the cervico-vaginal tissues could be analyzed by the TaqMan method if normalization to GAPDH mRNA was used and a significant L1-specific IFN- induction was found in 1/3 of immunized mice. This local response did not correlate to the immune responses measured in the periphery, with the exception of the sacral LN, a LN draining the genital mucosa, where a significant correlation was found. Our data show that the TaqMan method is sensitive enough to detect antigen-specific CD8 T cell responses in the genital mucosa of individual mice, and this may contribute to elaborate effective vaccines against genital pathogens.