Evaluation of cytokine gene expression in porcine spleen cells, peripheral blood mononuclear cells, and alveolar macrophages by competitive RT-PCR.

ABSTRACT

Cytokines act as an important regulator of immune responses. Since cytokine expression levels are generally very low, more accurate and reliable methods of measuring their expression are needed. In this study, a modified competitive reverse transcription-polymerase chain reaction assay was developed to determine the expression levels and patterns of porcine IFN-gamma, IL-2, IL-4, IL-10, IL-12 p 35, and IL-12 p40 in spleen cells, peripheral blood mononuclear cells (PBMC), and alveolar macrophages that were stimulated for 4 h by lipopolysaccharide or phytohemagglutinin. Of these cytokines, the expression level of IFN-gamma was the highest in all examined cells. Constitutive expression of IL-2 and IL-4 was demonstrated in spleen cells and PBMC stimulated with phytohemagglutinin. However, their expression extent was not determinable or extremely low in the lipopolysaccharide-stimulated spleen cells and alveolar macrophages. Moderately high IL-10 expression was observed in all examined cells. IL-12 p 35 expression in alveolar macrophages was always higher than in spleen cells and PBMC. IL-12 p40 expression in alveolar macrophages was higher than in PBMC, but was lower than in spleen cells. In spleen cells, the expression of IL-12 p40 was higher than that of IL-12 p 35. In alveolar macrophages and PBMC, however, IL-12 p 35 showed a higher expression than IL-12 p40. These results indicate that each cytokine has its own characteristic expression profile in different immune cells.