Pam2CSK4 induced immune tolerance against NS3 mediated inflammation.

<p>The microglial cells were treated with 50 ng/ml of Pam2CSK4 for 16 hrs. The medium was replaced with fresh growth medium with or without NS3 for another 6 hours. RT-PCR and ELISA was performed to detect cytokine gene and protein expression respectively. 6 hr NS3 exposed microglia served as positive control. <b>(A, B, C and D)</b> Pam2Csk3 treatment completely blocked IL-8, IL-6 and TNF-α expression. IL-1β expression was detected however the band intensities were less compared to that of the positive control. NC represents the PCR reagent negative control. <b>(E, F, G and H)</b> ELISA results shows that IL-6, TNF-α and IL-1β were secreted at a significant level in the Pam2CSK4 treated cells even after replacing the cells with fresh growth medium without agonist (# p < 0.05, ## p< 0.01). TNF-α and IL-1β were significantly up regulated in Pam2CSK4 + NS3 treated cells compared to Pam2CSK4 exposed cells (^ p < 0.05, ^^ p < 0.01). There was a significant down regulation for IL-8 and other 3 cytokines in the Pam2CSK4 + NS3 treated cells compared to NS3 alone treated cells (* p < 0.05, ** p < 0.01). PCR data is representative of 3 independent experiments. The data is expressed as mean (n = 3) ± SE.