Thy-1 is a differentiation antigen that characterizes immature murine erythroid and myeloid hematopoietic progenitors.
To determine the role of Thy-1 antigen in murine hematopoietic differentiation, bone marrow was treated with anti-Thy-1.2 antibody and complement or complement alone. Growth of immature hematopoietic progenitors, erythroid burst-forming units (BFU-E), and granulocyte/macrophage colony-forming units (CFU-GM) was greatly reduced following antibody and complement treatment and was not restored by mitogen-stimulated spleen cell supernatants. In contrast, more mature erythroid and myeloid progenitors, the erythroid colony-forming unit (CFU-E) and the macrophage progenitor stimulated by L-cell-conditioned media (LCM), were spared by anti-Thy-1.2 antibody and complement treatment. Here, to separate the effects of anti-Thy-1.2 antibody treatment on accessory cells from those on progenitors, splenic T cells and thymocytes were added to treated marrow at ratios of up to 200%. Growth of BFU-E and CFU-GM was not restored. To more precisely replace required accessory cells, male complement-treated marrow was cocultured with female anti-Thy-1.2 antibody and complement-treated marrow. Even marrow cells failed to restore female BFU-E and CFU-GM growth. Fluorescent-activated cell sorting (FACS) and immune sheep red cell rosetting with anti-Thy-1.2-labeled marrow were then performed to determine if immature hematopoietic progenitors bear Thy-1.2. These techniques revealed enrichment of BFU-E and CFU-GM in the Thy-1.2-positive fraction, demonstrating the presence of Thy-1.2 on early murine hematopoietic progenitors. CFU-E and CFU-M were present in the Thy-1.2-negative fraction following FACS separation. These data demonstrate that Thy-1.2 is a differentiation antigen, present on at least some murine BFU-E and CFU-GM and lost as they mature to CFU-E and CFU-M.
Miller
B A
BA
Lipton
J M
JM
Linch
D C
DC
Burakoff
S J
SJ
Nathan
D G
DG
D000818:Animals; D000954:Antigens, Surface / Q000032:analysis*; D001854:Bone Marrow Cells; D002478:Cells, Cultured; D003165:Complement System Proteins; D004912:Erythrocytes / Q000166:cytology / Q000276:immunology; D004920:Erythropoiesis; D005260:Female; D006098:Granulocytes / Q000166:cytology / Q000276:immunology; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology*; D007518:Isoantibodies; D007621:Karyotyping; D008264:Macrophages / Q000166:cytology / Q000276:immunology; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D012397:Rosette Formation; D013601:T-Lymphocytes; D018800:Thy-1 Antigens
D000954:Antigens, Surface; D007518:Isoantibodies; D018800:Thy-1 Antigens; C035678:anti-Thy antibody; D003165:Complement System Proteins