Interleukin-9 stimulates the proliferation of human myeloid leukemic cells
Interleukin-9 stimulates the proliferation of human myeloid leukemic cells
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DOI：
10.1182/blood.v87.9.3852.bloodjournal8793852
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发表时间：
1996-05-01
期刊：
影响因子：
20.3
通讯作者：
Tura, S
中科院分区：
文献类型：
--
作者：
Lemoli, RM;Fortuna, A;Tura, S
人白细胞介素-9（IL-9）刺激原始造血红细胞和多能祖细胞的增殖，以及选定的集落刺激因子（CSF）依赖性骨髓细胞系的生长。为了进一步阐明IL-9在急性白血病发展中的作用，我们评价了三种白血病细胞系和32例急性髓细胞白血病（AML）患者的原发性样本对重组人（rh）-IL-9单独和联合rh-IL-3、粒细胞-巨噬细胞CSF（GM-CSF）和干细胞因子（[SCF] c-kit配体）的增殖反应。HL 60，K562，和KG 1细胞和新鲜的AML细胞群体IL-9刺激后的集落形成能力进行了评估，而白血病样本的细胞周期特征，吖啶橙流式细胞术技术和溴脱氧尿苷（BRDU）掺入测定。此外，末端脱氧核苷酸转移酶测定（TDTA）和凝胶电泳DNA切割的标准分析用于评估IL-9诱导或预防细胞凋亡。IL-9作为单一细胞因子，在不同浓度下刺激三种骨髓细胞系在含血清和无血清条件下的殖民地形成，并且这种作用被抗IL-9单克隆抗体（MoAbs）完全消除。当对新鲜AML样品进行测试时，最佳浓度的IL-9导致所有研究病例中原始细胞集落形成的增加（平均值+/- SEM：19 +/- 10集落形成单位-白血病[CFU-L]/10（5）个细胞接种在对照培养物中，而107 +/- 32个细胞接种在IL-9补充的培养皿中，P <0.02）。IL-9对植物血凝素-淋巴细胞条件培养液（PHA-LCM）诱导的CFU-L有36.8%的促进作用，是本研究所用CSF（即SCF、IL-3和GM-CSF）中促进白血病细胞生长最有效的CSF。当T细胞耗尽的AML标本与浓度增加的细胞因子一起孵育时，也观察到IL-9的增殖活性。SCF与IL-9的联合应用对8例AML患者中的5例CFU-L生长有相加或协同作用（187 +/- 79集落对107 +/- 32 CFU-L，P = 0.05）。当IL-9与IL-3和GM-CSF组合时，也观察到正相互作用。AML样本的细胞周期分布研究表明，在大多数评估的病例中，单独IL-9显著增加了S期白血病细胞的数量。IL-9和SCF联合作用导致G（0）细胞比例显著降低（38.2% +/-24%vs58.6%+/-22%，P <0.05），并诱导G（1）和S期细胞增加。相反，IL-9单独或IL-9与SCF的组合对诱导或预防白血病细胞凋亡均无任何作用。总之，我们的研究结果表明，IL-9可能通过刺激白血病细胞进入S期而不是阻止细胞死亡在AML的发展中发挥作用。此外，IL-9与SCF协同作用，用于在细胞周期中募集静止的白血病细胞。(C)1996年，美国血液学会。
Human interleukin-9 (IL-9) stimulates the proliferation of primitive hematopoietic erythroid and pluripotent progenitor cells, as well as the growth of selected colony-stimulating factor (CSF)-dependent myeloid cell lines. To further address the role of IL-9 in the development of acute leukemia, we evaluated the proliferative response of three leukemic cell lines and 32 primary samples from acute myeloblastic leukemia (AML) patients to recombinant human (rh)-IL-9 alone and combined with rh-IL-3, granulocyte-macrophage CSF (GM-CSF), and stem cell factor ([SCF] c-kit ligand). The colony-forming ability of HL60, K562, and KG1 cells and fresh AML cell populations upon IL-9 stimulation was assessed by a clonogenic assay in methylcellulose, whereas the cell-cycle characteristics of leukemic samples were determined by the acridine-orange flow cytometric technique and the bromodeoxyuridine (BRDU) incorporation assay. In addition, the terminal deoxynucleotidyl transferase assay (TDTA) and standard analysis of DNA cleavage by gel electrophoresis were used to evaluate induction or prevention of apoptosis by IL-9. IL-9, as a single cytokine, at various concentrations stimulated the colony formation of the three myeloid cell lines under serum-containing and serum-free conditions, and this effect was completely abrogated by anti-IL-9 monoclonal antibodies (MoAbs). When tested on fresh AML samples, optimal concentrations of IL-9 resulted in an increase of blast colony formation in all the cases studied (mean +/- SEM: 19 +/- 10 colony-forming unit-leukemic [CFU-L]/10(5) cells plated in control cultures v 107 +/- 32 in IL-9-supplemented dishes, P < .02). IL-9 stimulated 36.8% of CFU-L induced by phytohemagglutinin-lymphocyte-conditioned medium (PHA-LCM), and it was the most effective CSF for promoting leukemic cell growth among those tested in this study (ie, SCF, IL-3, and GM-CSF). The proliferative activity of IL-9 was also observed when T-cell-depleted AML specimens were incubated with increasing concentrations of the cytokine. Addition of SCF to IL-9 had an additive or synergistic effect of the two cytokines in five of eight AML cases tested for CFU-L growth (187 +/- 79 colonies v 107 +/- 32 CFU-L, P = .05). Positive interaction was also observed when IL-9 was combined with IL-3 and GM-CSF. Studies of cell-cycle distribution of AML samples demonstrated that IL-9 alone significantly augmented the number of leukemic cells in S-phase in the majority of cases evaluated. IL-9 and SCF in combination resulted in a remarkable decrease of the G(0) cell fraction (38.2% +/- 24% v 58.6% +/- 22% of control cultures, P < .05) and induced an increase of G(1)- and S-phase cells. Conversely, neither IL-9 alone nor the combination of IL-9 and SCF had any effect on induction or prevention of apoptosis of leukemic cells. In summary, our results indicate that IL-9 may play a role in the development of AML by stimulating leukemic cells to enter the S-phase rather than preventing cell death. Moreover, IL-9 acts synergistically with SCF for recruiting quiescent leukemic cells in cell cycle. (C) 1996 by The American Society of Hematology.