Gunji Y, Nakamura M, Hagiwara T, Hayakawa K, Matsushita H, Osawa H, Nagayoshi K, Nakauchi H, Yanagisawa M, Miura Y
Department of Pediatrics, Jichi Medical School, Tochigi-ken, Japan.
Blood. 1992 Jul 15;80(2):429-36.
Advances in fluorescence-activated cell sorter technology have brought about multicolor analysis of cell phenotypes. To clarify the phenotypes of human hematopoietic stem cells (HSCs), we initially prepared novel antibodies against CD34 and labeled one of them (4A1) with allophycocyanin (APC). With this, we analyzed the phenotypes of CD34+ HSCs and showed that primitive HSCs or CD34+CD33- cells expressed adhesion molecules such as CD43, CD44, CD11a, CD11c, CD18, and leukocyte adhesion molecule (LAM-1). The more primitive hematopoietic cells or CD34+CD38- cells also expressed CD11a and CD18 with an incidence of 20% to 30%. To clarify the role of adhesion molecules in HSCs, we examined the colony forming capacity after long-term culture with allogeneic irradiated stromal layers. Among CD34+CD33- cells, CD18+ cells gave rise to colony-forming cells (CFCs) on stromal layers, but reached a maximum at week 2, after which the number of generated CFCs decreased. On the other hand, CD18- cells generated less CFCs than CD18+ cells at 2 to 3 weeks, but increased after 4 weeks of culture. When CD18 or CD11a antibody was added to a coculture system of CD34+CD33- cells with stromal layers, the number of generated CFCs decreased significantly compared with the no antibody control. Leukocyte function-associated antigen-1 (LFA-1) (CD11a/CD18) was expressed on some populations of hematopoietic cells and contributed to the proliferation by interacting with stromal cells. However, more primitive cells capable of reconstituting hematopoiesis did not express LFA-1. These data provide a rationale for the administration of anti-LFA-1 antibody after bone marrow transplantation for reducing the graft failure.
荧光激活细胞分选技术的进步带来了细胞表型的多色分析。为了阐明人类造血干细胞（HSC）的表型，我们最初制备了针对CD34的新型抗体，并将其中一种（4A1）用别藻蓝蛋白（APC）标记。借此，我们分析了CD34+HSC的表型，并表明原始HSC或CD34+CD33-细胞表达诸如CD43、CD44、CD11a、CD11c、CD18和白细胞粘附分子（LAM-1）等粘附分子。更原始的造血细胞或CD34+CD38-细胞也表达CD11a和CD18，发生率为20%至30%。为了阐明粘附分子在HSC中的作用，我们在用同种异体照射的基质层进行长期培养后检测了集落形成能力。在CD34+CD33-细胞中，CD18+细胞在基质层上产生集落形成细胞（CFC），但在第2周达到最大值，之后产生的CFC数量减少。另一方面，CD18-细胞在2至3周时产生的CFC比CD18+细胞少，但在培养4周后增加。当将CD18或CD11a抗体添加到CD34+CD33-细胞与基质层的共培养系统中时，与无抗体对照相比，产生的CFC数量显著减少。白细胞功能相关抗原-1（LFA-1）（CD11a/CD18）在一些造血细胞群体上表达，并通过与基质细胞相互作用促进增殖。然而，能够重建造血的更原始细胞不表达LFA-1。这些数据为骨髓移植后给予抗LFA-1抗体以减少移植物失败提供了理论依据。
