Generation of functional human dendritic cells from adherent peripheral blood monocytes by CD40 ligation in the absence of granulocyte-macrophage colony-stimulating factor
Generation of functional human dendritic cells from adherent peripheral blood monocytes by CD40 ligation in the absence of granulocyte-macrophage colony-stimulating factor
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DOI：
10.1182/blood.v92.11.4238.423k16_4238_4247
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发表时间：
1998-12-01
期刊：
影响因子：
20.3
通讯作者：
Brugger, WR
中科院分区：
文献类型：
--
作者：
Brossart, P;Grünebach, F;Brugger, WR
近年来研究表明，在粒细胞-巨噬细胞集落刺激因子（GM-CSF）和白细胞介素-4（IL-4）存在下，外周血单核细胞可分化为树突状细胞（DC）。然而，目前尚不清楚DC是否也可以在缺乏这些细胞因子的情况下从单核细胞发育而来。因此，我们分析了Flt-3配体（Flt 3L）和CD 40配体对在不存在GM-CSF的情况下从血液单核细胞发育人DC的作用。在存在不同细胞因子组合的情况下培养贴壁外周血单核细胞（PBMNC），并分析表面分子的表达和抗原呈递能力。对于功能分析，测试细胞在混合淋巴细胞反应（MLR）中刺激同种异体T淋巴细胞、呈递可溶性抗原和体外诱导原代HIV肽特异性细胞毒性T细胞（CTL）应答的能力。此外，DC-CK 1，最近确定的DC中的特异性表达的趋化因子，IL-18（IGIF），Th 1淋巴细胞的生长和分化因子的表达，通过逆转录聚合酶链反应（RT-PCR）进行了分析。在我们的研究中，单独的Flt 3L不足以产生DC，并且需要添加IL-4。用Flt 3L和IL-4产生的DC在用肿瘤坏死因子-α（TNF-α）或CD 40 L刺激后经历成熟，其特征在于CD 83表达、MHC、粘附和共刺激分子的上调以及同种异体增殖反应的增加。相比之下，在缺乏GM-CSF和IL-4的情况下，单独的CD 40连接促进粘附的血液单核细胞分化为功能性DC。这些细胞表现出成熟DC的所有表型和功能特征，并且在针对HIV肽的主要组织相容性复合体（MHC）I类限制性CTL应答的引发中是有效的刺激细胞，而它们呈递可溶性蛋白抗原的能力降低。使用半定量RT-PCR，在所有产生的DC群体中检测到DC-CK 1和IL-18转录物，与所使用的生长因子无关。我们的研究结果进一步证明了CD 40-CD 40 L相互作用对启动和维持T细胞应答的重要性，并证实了血液单核细胞依赖于外部刺激提供DC的额外来源这一新兴概念。(C)1998年，美国血液学会。
Recently it has been shown that dendritic cells (DC) can develop from peripheral blood monocytes when grown in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). However, it is unclear whether DC can also develop from monocytes in absence of these cytokines. We therefore analyzed the effect of Flt-3 ligand (Flt3L) and of CD40 ligand on the development of human DC from blood monocytes in the absence of GM-CSF Adherent peripheral blood mononuclear cells (PBMNC) were cultured in the presence of different cytokine combinations and analyzed for the expression of surface molecules and antigen presenting capacity. For functional analyses, cells were tested for their ability to stimulate allogeneic T lymphocytes in a mixed lymphocyte reaction (MLR), to present soluble antigens, and to induce primary HIV-peptide-specific cytotoxic T-cell (CTL) responses in vitro. Furthermore, expression of DC-CK1, a recently identified chemokine with specific expression in DC, and of IL-18 (IGIF), a growth and differentiation factor for Th 1 lymphocytes, was analyzed by reverse-transcription polymerase chain reaction (RT-PCR). In our study, Flt3L alone was not sufficient to generate DC and required addition of IL-4. DC generated with Flt3L and IL-4 underwent maturation after stimulation with tumor necrosis factor-alpha (TNF-alpha) or CD40L, characterized by CD83 expression, upregulation of MHC, adhesion, and costimulatory molecules as well as increased allogeneic proliferative response. In contrast, CD40 ligation alone promoted differentiation of adherent blood monocytes into functional DC in the absence of GM-CSF and IL-4. These cells displayed all phenotypic and functional characteristics of mature DC and were potent stimulatory cells in priming of major histocompatibility complex (MHC) class I-restricted CTL responses against an HIV-peptide, whereas their ability to present soluble protein antigens was reduced. Using a semiquantitative RT-PCR, DC-CK1 and IL-18 transcripts were detected in all generated DC populations, independent of growth factors used. Our findings provide further evidence for the importance of CD40-CD40L interaction for initiation and maintenance of T-cell responses and confirm the emerging concept that blood monocytes provide an additional source of DC depending on external stimuli. (C) 1998 by The American Society of Hematology.