Saito Yoshinobu, Guo Yong Mei, Hirokawa Makoto, Saito Kunie, Komatsuda Atsushi, Takahashi Naoto, Fujishima Masumi, Fujishima Naohito, Yamashita Junsuke, Sawada Kenichi
Department of Internal Medicine III, Akita University School of Medicine, Hondo 1-1-1, Akita, 010-8543, Japan.
Radioisotope Division, Bioscience center, Akita University Graduate School of Medicine, Akita, 010-8543, Japan.
Int J Hematol. 2008 Jul;88(1):64-72. doi: 10.1007/s12185-008-0098-z. Epub 2008 May 20.
Tumor necrosis factor-alpha (TNF-alpha) has been shown to induce the differentiation of CD34(+) cells toward dendritic cells (DCs). We have previously shown that DCs are co-generated from human CD34(+) cells during erythroid or megakaryocytic differentiation in the presence of TNF-alpha, and those DCs are able to stimulate autologous T cell proliferation. The aim of this study was to learn whether the co-stimulation of granulocyte colony-stimulating factor (G-CSF) and TNF-alpha would generate neutrophil progenitors and DCs together from human CD34(+) cells, and if this was the case, to clarify the phenotypic and functional characteristics of these DCs. When highly purified human CD34(+) cells were cultured for 7 days with G-CSF alone, the generated cells predominantly expressed a granulocyte marker, CD15, and then differentiated into neutrophils after 14 days of culture. The addition of TNF-alpha with G-CSF markedly decreased the number of CD15(+) cells without affecting the total number of cells during 7 days of culture. Almost one third of the generated cells were positive for CD11c and CD123. Furthermore, CD11c(+) cells were found to phagocytose CD15(+) cells and were able to induce allogeneic, but not autologous, T cell proliferation in the mixed lymphocyte reaction (MLR). On the other hand, the CD11c(+) cells generated by TNF-alpha and cytokines capable of inducing erythroid differentiation were able to stimulate autologous T cells. There was a difference in the expression of CD80, CD83 and CD86 among CD11c(+) cells induced by G-CSF plus TNF-alpha and those generated by interleukin-3, stem cell factor, and erythropoietin plus TNF-alpha. These results indicate that the co-stimulation of human CD34(+) cells with G-CSF and TNF-alpha induces the phagocytosis of co-developing neutrophil progenitors by DCs, and the stimulatory effects of these DCs on autologous T cells is different from that of DCs generated from CD34(+) cells during erythroid differentiation.
肿瘤坏死因子-α（TNF-α）已被证明可诱导CD34(+)细胞向树突状细胞（DCs）分化。我们之前已经表明，在TNF-α存在的情况下，DCs在人类CD34(+)细胞进行红系或巨核系分化过程中共同生成，并且这些DCs能够刺激自体T细胞增殖。本研究的目的是了解粒细胞集落刺激因子（G-CSF）和TNF-α的共同刺激是否会从人类CD34(+)细胞中共同产生中性粒细胞祖细胞和DCs，如果是这样，阐明这些DCs的表型和功能特征。当高度纯化的人类CD34(+)细胞单独用G-CSF培养7天时，所生成的细胞主要表达粒细胞标志物CD15，然后在培养14天后分化为中性粒细胞。在培养7天期间，G-CSF与TNF-α一起添加显著减少了CD15(+)细胞的数量，但不影响细胞总数。几乎三分之一的所生成细胞CD11c和CD123呈阳性。此外，发现CD11c(+)细胞吞噬CD15(+)细胞，并且能够在混合淋巴细胞反应（MLR）中诱导异体而非自体T细胞增殖。另一方面，由TNF-α和能够诱导红系分化的细胞因子所生成的CD11c(+)细胞能够刺激自体T细胞。G-CSF加TNF-α诱导生成的CD11c(+)细胞与白细胞介素-3、干细胞因子以及促红细胞生成素加TNF-α诱导生成的CD11c(+)细胞在CD80、CD83和CD86的表达上存在差异。这些结果表明，G-CSF和TNF-α对人类CD34(+)细胞的共同刺激诱导DCs吞噬共同发育的中性粒细胞祖细胞，并且这些DCs对自体T细胞的刺激作用与红系分化过程中从CD34(+)细胞生成的DCs的刺激作用不同。
