Th1/Th2 biasing effects of vaccination in cattle as determined by real-time PCR.

Real-time polymerase chain reaction (PCR) is now becoming an accepted tool for measuring gene expression at the transcriptional level. In this study, a direct comparison between real-time PCR, enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunosorbent spot (ELISPOT) assay was performed. When interferon-gamma (IFN-gamma) gene expression was assessed, both ELISA and ELISPOT data strongly correlated to results obtained by real-time PCR. Real-time PCR was subsequently used to measure bovine IFN-gamma (bIFN-gamma) and bovine interleukin-4 (bIL-4) gene expression by antigen stimulated peripheral blood mononuclear cells (PBMC), isolated from bovine herpesvirus-1 (BHV-1) infected animals. BHV-1-infected animals were either non-vaccinated or vaccinated using one of two adjuvants prior to infection. With non-vaccinated infected animals, a Th1 bias occurred, based on IFN-gamma expression exceeding IL-4 expression. The level of cytokine expression, and the IFN-gamma/IL-4 ratio could be significantly affected, depending on the manner in which animals were vaccinated.