Differential cytokine mRNA expression in swine whole blood and peripheral blood mononuclear cell cultures

The kinetics of interleukin-2 (IL-2), IL-6, IL-8 and IL-10 gene expression in concanavalin A (Con A)-activated whole blood (WB) and peripheral blood m ononuclear cell (PBMC) cultures were examined using reverse transcriptase-p olymerase chain reaction (RT-PCR). Unstimulated PBMC or WE cultures failed to show increases in basal cytokine PCR amplicon levels for any cytokine ex amined. PBMC cultures demonstrated peak expression of IL-2, IL-6, IL-8 and IL-10 mRNA levels at 12, 24, 24 and 6 h, respectively. WE cultures exhibite d peak IL-2, IL-6, IL-8 and IL-10 mRNA levels at 24, 12, 6 and 24 h, respec tively. PBMC cultures consistently exhibited higher levels of IL-2 mRNA at all times examined than did WE cultures. WE cultures consistently had highe r levels of IL-6 mRNA than PBMC cultures. IL-8 and IL-10 protein levels in PBMC cultures were first detected 12 h after stimulation and continued to i ncrease in concentration through 48 h. In WE cultures, 1L-8 and IL-10 prote in levels were first noted at 12 and 6 h, respectively. WE culture IL-8 and IL-10 levels quickly reached equilibrium after being detected and remained at levels lower than those noted in PBMC cultures. These results show WE c ultures represent an approach with reduced cost and time when compared to t raditional cell culture and isolation methods. It may also produce an in vi tro test system that more closely resembles in vivo conditions. Published b y Elsevier Science B.V.