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Objective: Hepatitis C virus is a main cause of progressive and chronic liver disease such as hepatitis, cirrhosis and hepatocellular carcinoma which resulted in global health concern. There are 170 million people infecting HCV in the world. Autophagy is a catabolic process 'degrading damaged organelles and cellular components for recycling. There are conflict reports about the role of autophagy in production of cytokines in viral infection. It may depend on the type of virus and the type of infected cells. However, it has been reported that in HCV infected cells, dysfunction of autophagy led to enhancement in some products of native immunity factors such as interferon-α. HCV has been shown to induce autophagy to increase viral growth in vitro. Pharmacological inhibition of autophagic process remarkably inhibited expression level of HCV replicon. In this study we examined the impact of autophagy inhibition by chloroquine'a lysosomal acidification inhibitor'on IFNα mRNA expression and HCV replication in PBMCs. Material and Methods: Immediately after blood collection from HCV positive patients who have chronic HCV genotype1a before treatment the PBMCs were separated from whole blood by centrifugation on a Ficoll-Hypaque density gradient. The same number of normal samples were tested as controls. The cells were cultured in RPMI medium and incubated with 50μgr chloroquine. MTT assay was applied to indicate the dose of chloroquine that has minimum cytotoxicity for cells. After 72h, viral RNA was extracted. The real-time PCR was carried out after a reverse transcription step. The result had been analyzed by REST software. Results: The use of CQ for inhibition of autophagy in infected PBMCs with HCV resulted in a decrease amount HCV mRNA with an increase in IFNα mRNA expression. These results indicated that chloroquine may consider as a complementary new anti-HCV agent that targets the autophagic proteolysis. Conclusion: Inhibition of autophagy may be a new therapeutic strategy for HCV treatment. CQ (chloroquine), inhibits autophagy in infected PBMCs and led to increase of IFNα mRNA expression. This increase has an inhibitory effect on HCV replication. These findings imply that chloroquine effectively impairs the function of autophagy in our experiment. [ABSTRACT FROM AUTHOR]