Pam3CSK4 induced immune tolerance against HCV NS3 mediated inflammation.

Microglia was pre treated with 50 ng/ml of Pam3CSK4 for 16 hrs. RT-PCR and ELISA was performed to study the immune tolerance mediated by Pam3CSK4. (A, B, C and D) Pam3CSK4 treatment completely blocked IL-8, IL-6 and TNF-α gene expression and IL-1β gene expression was down regulated compared to positive control (PC NC represents the PCR reagent negative control. (E, F, G and H) TNF-α and IL-1β were secreted at a significant level in the Pam3CSK4 treated cells even after replacing the cells with fresh growth medium without agonist (## p< 0.01). TNF-α and IL-1β were significantly upregulated in Pam3CSK4 + NS3 treated cells compared to Pam3CSK4 exposed cells (^ p < 0.05, ^^ p < 0.01). There was a significant down regulation for IL-8 and other 3 cytokines in the Pam3CSK4 + NS3 treated cells compared to NS3 alone treated cells (** p < 0.01). The data is expressed as mean (n = 3) ± SE.