The ability of HTLV-I to infect cells is presumed to be dependent, in some part, on the attachment of the virus to a target cell via a specific cell surface receptor which is, as yet, unknown. Here we present evidence that a monoclonal antibody, Mab 34-23, inhibits the binding of HTLV-I to IL-2 and phytohemagglutinin-activated peripheral blood mononuclear cells and also inhibits virus entry into these cells. Analysis of a variety of target cells, including a human:mouse somatic hybrid which contains only human chromosome 17q, indicates that the binding of Mab 34-23 correlates with HTLV-I adsorption and entry. SDS-PAGE and Western blot analysis show that Mab 34-23 binds to four major proteins of MW 31, 45, 55, and 70 kDa and this binding can be inhibited by HTLV-I and not HIV proteins. HTLV-I virions bind to proteins of similar molecular weight and virus-binding to these proteins can be inhibited by preincubation with Mab 34-23. These data suggest that Mab 34- 23 may identify a specific cell surface receptor(s) for HTLV-I. © 1993 American Health Foundation and Academic Press.