Macrophages constitute a major component in human hepatocellular carcinoma (HCC) and perform various functions to facilitate disease progression. Reprogramming or reconstituting the tumor surveillance phenotypes of macrophages represents an attractive immunotherapeutic strategy in cancer treatments. The current study identified CD169 as potential target for macrophages repolarization since it signified a population of macrophages positively correlated with activated immune signature and better prognosis of patients with HCC. In vitro experiments revealed that low-dose of type I IFN could effectively reprogram human monocyte-derived macrophages to up-regulate CD169 expression, and such induced CD169+ macrophages exhibited significantly enhanced phagocytotic and CD8+ T cell -activating capacities compared to control. Low dose of IFNα also inhibited hepatoma growth in mice in vivo, presumably through polarizing the CD169+ macrophage population and enhancing CD8+ T cell activities. Notably, IFNα also induced substantial PD-L1 expression on macrophages in vivo, thus blockade of PD-L1 could further increase the anti-tumor efficacy of IFNα in the treatment of HCC. Together, we proposed low-dose of IFNα in combination with PD-L1 blocking agent as a potential anti-tumor therapeutic strategy via its effects on macrophages polarization.Copyright © 2021 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.