Fukaya Hiroshi, Xiao Weiguo, Inaba Kayo, Suzuki Yoshiko, Hirokawa Makoto, Kawabata Yoshinari, Komatsuda Atsushi, Endo Tomoyuki, Kishimoto Hiroyuki, Takada Goro, Sawada Kenichi
Department of Animal Development and Physiology, Graduate School of Biostudies, Kyoto University, Kyoto, Japan.
Exp Hematol. 2004 May;32(5):450-60. doi: 10.1016/j.exphem.2004.02.011.
Tumor necrosis factor-alpha (TNF-alpha) inhibits erythropoiesis and enhances nonerythroid colony formation. The present study examines the nature of these nonerythroid cells and investigates their physiologic role in relation to erythroid progenitor cells.
Highly purified human CD34(+) cells underwent erythroid differentiation in the presence of multiple cytokines, including stem cell factor (SCF), interleukin-3 (IL-3), and erythropoietin (EPO), with and without TNF-alpha. We enumerate colony-forming unit-erythroid (CFU-E) and glycophorin A (GPA; a specific marker for erythroid lineage) positive cells in semisolid phase as well as in liquid suspension culture. The character and roles of codeveloping nonerythroid cells in the presence of TNF-alpha were analyzed using fluorescent activating cell sorter, enzyme immunohistochemistry, and confocal microscopy.
TNF-alpha inhibited the generation of GPA(+) cells and conversely enhanced the generation of GPA(-) cells. The GPA(-) cells were comprised of cells with excentric cell shape and were positive for HLA class I, HLA class II, CD1a, CD4, CD11c, CD14, CD40, CD80, CD83, and CD86, but not for CD3, CD8, CD19, CD20, and CD56, indicating the codevelopment of dendritic cells (DC) along with erythroid differentiation. Developing DC/DC precursors were detected within 3 days of culture. Only in the presence of TNF-alpha did CD34(+) cells proliferate by forming aggregates where both GPA(+) and CD11c(+) DC/DC precursors were present. During culture period, immature CD11c(+) DC were capable of endocytosing damaged GPA(+) cells.
GPA(-) cells cogenerated from human CD34(+) cells during erythroid differentiation in the presence of IL-3/SCF/EPO and TNF-alpha express DC phenotypes. The CD11c(+) DC subset physically and selectively associates with developing immature erythroid cells and damaged self-GPA(+) cells and then obtains and captures self-substances.
肿瘤坏死因子-α（TNF-α）抑制红细胞生成并增强非红系集落形成。本研究探讨这些非红系细胞的性质，并研究它们与红系祖细胞相关的生理作用。
高度纯化的人CD34(+)细胞在多种细胞因子存在的情况下进行红系分化，这些细胞因子包括干细胞因子（SCF）、白细胞介素-3（IL-3）和促红细胞生成素（EPO），同时加入或不加入TNF-α。我们在半固体阶段以及液体悬浮培养中对红系集落形成单位（CFU-E）和血型糖蛋白A（GPA；红系谱系的特异性标志物）阳性细胞进行计数。使用荧光激活细胞分选仪、酶免疫组织化学和共聚焦显微镜分析在TNF-α存在下共同发育的非红系细胞的特征和作用。
TNF-α抑制GPA(+)细胞的生成，相反增强了GPA(-)细胞的生成。GPA(-)细胞由细胞形状偏心的细胞组成，对I类 HLA、II类 HLA、CD1a、CD4、CD11c、CD14、CD40、CD80、CD83和CD86呈阳性，但对CD3、CD8、CD19、CD20和CD56呈阴性，表明树突状细胞（DC）与红系分化共同发育。在培养3天内检测到正在发育的DC/DC前体。仅在TNF-α存在的情况下，CD34(+)细胞通过形成聚集物而增殖，其中同时存在GPA(+)和CD11c(+) DC/DC前体。在培养期间，未成熟的CD11c(+) DC能够内吞受损的GPA(+)细胞。
在IL-3/SCF/EPO和TNF-α存在的情况下，人CD34(+)细胞在红系分化过程中共同产生的GPA(-)细胞表达DC表型。CD11c(+) DC亚群在物理上与正在发育的未成熟红系细胞以及受损的自身GPA(+)细胞选择性结合，然后获取并捕获自身物质。
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