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.
Mena
Angelo
A
Veterinary Infectious Disease Organisation, University of Saskatchewan, Saskatoon, Canada. angelo.mena@sask.usask.ca
Ioannou
Xenia P
XP
Van Kessel
Andrew
A
Van Drunen Little-Van Den Hurk
Sylvia
S
Popowych
Yurij
Y
Babiuk
Lorne A
LA
Godson
Dale L
DL
D000818:Animals; D002417:Cattle; D015870:Gene Expression; D006566:Herpesviridae Infections / Q000235:genetics / Q000276:immunology*; D007242:Herpesvirus 1, Bovine / Q000276:immunology*; D007371:Interferon-gamma / Q000235:genetics; D015847:Interleukin-4 / Q000235:genetics; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods; D018417:Th1 Cells / Q000276:immunology*; D018418:Th2 Cells / Q000276:immunology*; D014611:Vaccination
D015847:Interleukin-4; D007371:Interferon-gamma