Abstract The precise cell type hosting latent human cytomegalovirus (HCMV) remains elusive. Here we report that HCMV reprograms human hematopoietic progenitor cells (HPCs) into a unique monocyte subset to achieve latency. Unlike conventional monocytes, this monocyte subset possesses higher levels of B7-H4, IL-10 and iNOS, longer lifespan and strong immunosuppressive capacity. Cell sorting of peripheral blood from latently infected human donors confirms that only this monocyte subset, representing less than 0.1% of peripheral mononuclear cells, is HCMV genome-positive but immediate-early (IE)-negative. Mechanistic studies demonstrate that HCMV promotes the differentiation of HPCs into this monocyte subset by activating cellular signal transducer and activator of transcription 3 (STAT3). In turn, this monocyte subset generates high level of nitric oxide (NO) to silence HCMV IE transcription and promote viral latency. By contrast, US28-knockout HCMV mutant incapable of activating STAT3, fails to reprogram the HPCs and achieve latency. Our findings reveal that HCMV differentiates human HPCs into long-life, immunosuppressive monocyte subset for viral latency via activating STAT3/iNOS/NO axis.