Th1 and Th2 cytokine gene expression in primary infection and vaccination against Fasciola gigantica in buffaloes by real-time PCR

Abstract

Th1 and Th2 cytokine gene expression in buffalo calves during primary infection with Fasciola gigantica as well as in response to immunization with the parasite recombinant fatty acid binding protein (rFABP) and recombinant glutathione S-transferase (rGST) proteins was measured at 14th week of infection by real-time PCR with the double-stranded DNA-binding dye SYBR Green. Experimental animals were randomly distributed into FABP, GST, cocktail, challenge and healthy groups. Animals in groups FABP and GST were immunized with 400 μg rFABP and rGST, respectively, and cocktail group with a mixture of 400 μg each of rFABP and rGST in the neck and thigh muscles. All animals received three immunizations at 3-week interval. Calves were challenged per os with 400 viable metacercariae along with the unimmunized challenge control group 1 month after the last immunization. Expression of various cytokines in response to the immunization and parasite primary infection was measured by real-time PCR. Expression of IL-2 (4.5-fold) and IFN-γ (3.2-fold), followed by IL-6 (1.7-fold) and IL-4 (1.6-fold), with downregulation of TNF-α and IL-10 was observed in response to F. gigantica infection in these animals. However, there was a sharp increase in the expression of the IL-4 (211.93 and 111.81-fold) and IL-6 mRNA (219.22 and 48.29-fold) to GST and FABP immunizations, respectively. A downregulation of the IL-1α, a Th1 cytokine in response to FABP and GST immunization in these calves, was also observed. Overall, a mixed type of Th1 and Th2 cytokine environment was evoked to chronic F. gigantica infection and immunization with the above two recombinant proteins in buffaloes.