Thrombocytopenia is a common hematologic disorder in HIV infection and occurs in both asymptomatic and AIDS patients. An autoimmune mechanism has been postulated for the platelet destruction associated with some forms of thrombocytopenia. However, recent studies revealed that megakaryocytes are susceptible to HIV infection and suggested the possibility that HIV can directly impair the platelet production from megakaryocytes. This study was designed to characterize the HIV receptor expression in megakaryocytic cells and the responsiveness to HIV infection. Four different megakaryocytic cell lines at different stages of differentiation were established from the peripheral blood of different individuals with hematologic malignancies. CMK and CMY cells (differentiated cell lines) expressed CD4, but CMS and CTS cells (poorly differentiated cell lines) did not. The HIV coreceptor CXCR4 was also expressed in CMY and CMK cells. HIV-1 (HTLV-IIIB) replicated in CMY cells persistently but not in other three cell lines. CMY cells as well as CMK cells were also susceptible to the lytic infection of HIV-2 (LAV2). Pretreatment of the CMY cells with anti-CD4 antibody inhibited the infection by both HIV-1 and HIV-2. Our results indicate that mature megakaryocytic cells express CD4 along with HIV coreceptors and are susceptible to HIV infection.