Miller B A, Lipton J M, Linch D C, Burakoff S J, Nathan D G
J Cell Physiol. 1985 Apr;123(1):25-32. doi: 10.1002/jcp.1041230105.
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.
为了确定Thy-1抗原在小鼠造血分化中的作用，用抗Thy-1.2抗体和补体或仅用补体处理骨髓。抗体和补体处理后，未成熟造血祖细胞、红系爆式集落形成单位（BFU-E）和粒细胞/巨噬细胞集落形成单位（CFU-GM）的生长显著降低，且丝裂原刺激的脾细胞上清液不能使其恢复。相反，更成熟的红系和髓系祖细胞，即红系集落形成单位（CFU-E）和由L细胞条件培养基（LCM）刺激的巨噬细胞祖细胞，不受抗Thy-1.2抗体和补体处理的影响。在此，为了区分抗Thy-1.2抗体处理对辅助细胞和祖细胞的影响，将脾T细胞和胸腺细胞以高达200%的比例添加到处理过的骨髓中。BFU-E和CFU-GM的生长未恢复。为了更精确地替代所需的辅助细胞，将雄性补体处理的骨髓与雌性抗Thy-1.2抗体和补体处理的骨髓共培养。即使是骨髓细胞也未能恢复雌性BFU-E和CFU-GM的生长。然后进行荧光激活细胞分选（FACS）和用抗Thy-1.2标记的骨髓进行免疫羊红细胞花环试验，以确定未成熟造血祖细胞是否携带Thy-1.2。这些技术显示BFU-E和CFU-GM在Thy-1.2阳性部分富集，表明早期小鼠造血祖细胞上存在Thy-1.2。FACS分离后，CFU-E和CFU-M存在于Thy-1.2阴性部分。这些数据表明，Thy-1.2是一种分化抗原，至少存在于一些小鼠BFU-E和CFU-GM上，并在它们成熟为CFU-E和CFU-M时丢失。
