The Monoclonal Antibody W7C5 Defines a Novel Surface Antigen on Hematopoietic Stem Cells

Abstract

We recently raised a monoclonal antibody, termed W7C5, against a surface antigen that is expressed at low levels on bone marrow and peripheral blood CD34+ stem/progenitor cells but at high levels on fetal liver CD34+ cells. A reasonable staining intensity was achieved using magnetofluorescent liposome conjugates to analyze expression of W7C5 antigen on CD34+CD38- bone marrow (BM) cells. Flow cytometric analyses revealed that W7C5 detects about 50% of immature CD34+CD38- BM cells that coexpressed the differentiation antigens CD164, CD133, and CD172a (SIRP alpha). In addition, W7C5 also recognized a CD34- BM fraction. These cells were negative for CD117 and CD133, but expressed CD45 and moderate levels of CD164. Injection of selected CD34+W7C5+ and CD34-W7C5+ cells into 55-60-day-old fetal sheep resulted in an engraftment of both fractions. Partial amino acid sequence analysis of affinity-purified lysates of KU-812 cells revealed that W7C5 detects a novel membrane protein. Together, W7C5 defines a novel molecule that is expressed on CD34+ as well as on CD34- stem cell subsets.