Toll-Like Receptor 7 (TLR-7) and TLR-9 Agonists Improve Hepatitis C Virus Replication and Infectivity Inhibition by Plasmacytoid Dendritic Cells.

Plasmacytoid dendritic cells (pDCs) are innate immune cells with high antiviral activity triggered by Toll-like receptor 7 (TLR-7) and TLR-9 stimulation. Moreover, they are important mediators between innate and adaptive immunity. Although nowadays there is available an effective therapeutic arsenal against hepatitis C virus (HCV), a protective vaccine is not available. We have analyzed the pDCs' response to HCV infection in a hepatitis C virus (HCV)-Huh7.5 virus-cell system, which allows completion of the virus infectious cycle. pDCs were cocultured following human immunodeficiency virus (HIV) aldrithiol-2 (AT-2 [TLR-7 agonist]) inactivation and CpG (TLR-9 agonist) stimulation. We employed three virus derivatives-wild-type Jc1, interferon (IFN)-resistant virus IR, and high-replicative-fitness virus P100-in order to explore additional IFN-α-related virus inhibition mechanisms. pDCs inhibited HCV infectivity and replication and produced IFN-α. After TLR-7 and TLR-9 stimulation, inhibition of infectivity and IFN-α production by pDCs were enhanced. TLR-7 stimulation drove higher TNF-related apoptosis-inducing ligand (TRAIL) expression in pDCs. Additionally, TLR-7- and TLR-9-stimulated pDCs exhibited a mature phenotype, improving the antigen presentation and lymph node homing-related markers. In conclusion, pDCs could serve as a drug target against HCV in order to improve antiviral activity and as an enhancer of viral immunization.IMPORTANCE We implemented a coculture system of pDCs with HCV-infected hepatoma cell line, Huh7.5. We used three HCV derivatives in order to gain insight into pDCs' behavior against HCV and associated antiviral mechanisms. The results with this cell coculture system support the capacity of pDCs to inhibit HCV replication and infectivity mainly via IFN-α, but also through additional mechanisms associated with pDC maturation. We provided evidence that TLR agonists can enhance antiviral pDCs' function and can induce phenotypic changes that may facilitate the interplay with other immune cells. These findings suggest the possibility of including TLR agonists in the strategies of HCV vaccine development.

Pneumocystis carinii induces interleukin 6 production by an alveolar epithelial cell line.

BACKGROUND: Pneumocystis carinii (PC) infection often results in severe pneumonia in immunocompromised patients. Attachment of PC organisms to alveolar epithelial cells is a hallmark of PC pneumonia; however, few studies have investigated the response of alveolar epithelial cells to PC infection. METHODS: Interleukin 6 (IL-6) is a multifunctional cytokine that is produced by alveolar epithelial cells in response to a variety of stimuli. Our investigation was undertaken to determine whether PC organisms induce production of IL-6 by alveolar epithelial cells and to determine the effect of IL-6 on PC attachment. RESULTS: Incubation of the human alveolar epithelial cell line, A549, with PC organisms resulted in a significant increase in IL-6 secreted into the cell culture media. Time-course studies showed that IL-6 production was detected as soon as 2 h after addition of PC and continued up to 48 h of exposure. Further studies demonstrated that preincubation of A549 cells with IL-6 resulted in a concentration-dependent increase in both A549 cell production of fibronectin and PC attachment. CONCLUSION: Thus, PC attachment to an alveolar epithelial cell line results in epithelial cell production of IL-6, which can act to further increase PC attachment. This may provide an important mechanism whereby PC organisms directly affect the host response to PC infection.