Figure 4

scSIV expressing LMP1 or LMP1-CD40 induces increased secretion of inflammatory cytokines and β-chemokines. Human inflammatory cytokine quantitation was performed by cytometric bead array (CBA). Cytokine concentrations from three independent experiments are presented. Data were analyzed with the unpaired t test: *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared with the scSIV-GFP infected group. MIP-1α, MIP-1β, RANTES mRNA expression was analyzed by real-time RT-PCR assay. (A) DCs were infected with the various SIV viruses at MOI of 0.05 and supernatants were collected at various time intervals. Virus expressing LMP1 resulted in a significant increase in IL-1β, IL-6, IL-8, IL-10, IL-12p70 and TNF, while LMP1-CD40 resulted in an increase in IL-1β, IL-6, IL-8, IL-10 and TNF at various time points post infection. No measurable amounts of IL-12p70 were detected in GFP and LMP1-CD40 groups. (B) Macrophages were infected with different scSIV viruses for 4 days. Total cellular RNA was isolated, reverse transcribed to cDNA and MIP-1α, MIP-1β, RANTES mRNA expression was analyzed by real-time PCR assay. Virus expressing LMP1 resulted in significant increase in MIP-1β and RANTES mRNA expression, whereas LMP1-CD40 resulted in significant increase in MIP-1α, MIP-1β and RANTES mRNA expression. Expression of GAPDH was used for normalization of samples.