Bacterial lipopolysaccharide stimulates the production of cytokines and the expression of costimulatory molecules by human peripheral blood dendritic cells: evidence for a soluble CD14-dependent pathway.
Bacterial lipopolysaccharide stimulates the production of cytokines and the expression of costimulatory molecules by human peripheral blood dendritic cells: evidence for a soluble CD14-dependent pathway.
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DOI：
10.4049/jimmunol.158.6.2919
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发表时间：
1997-03
影响因子：
4.4
通讯作者：
V. Verhasselt;C. Buelens;F. Willems;D. Groote;N. Haeffner‐cavaillon;M. Goldman
中科院分区：
文献类型：
--
作者：
V. Verhasselt;C. Buelens;F. Willems;D. Groote;N. Haeffner‐cavaillon;M. Goldman
为了研究革兰氏阴性菌感染过程中树突状细胞（DC）的反应，我们分析了不同剂量LPS对IL-4和粒细胞-巨噬细胞-CSF存在下培养塑料贴壁PBMC产生的人DC的细胞因子谱、表型和同种异体刺激潜能的影响。首先，我们发现LPS刺激DC产生高水平的TNF-α、IL-6、IL-8、IL-12，并上调其HLA-DR、B7-1、B7-2和CD 40的表达。LPS的作用是剂量依赖性的，在0.1 ng/ml的浓度下已经观察到显著的刺激作用，在10 ng/ml时达到平台。这些表型变化与LPS激活的DC的增加的同种异体刺激特性相关，因为用LPS处理的DC在通过同种异体反应性T细胞诱导IL-2和IFN-γ合成并刺激其增殖方面比未处理的DC显著更有效。使用中和性抗IL-12 mAb的实验表明，LPS诱导的IL-12是MLR期间IFN-γ产生增加的原因，但不是MLR期间增殖增加的原因。最后，我们观察到DC对低水平LPS（1 ng/ml）的反应被阻断性抗CD 14 mAb显著抑制，尽管DC在其膜上不表达CD 14分子。使用去除可溶性CD 14（sCD 14）的血清和从人血清纯化的或重组形式的sCD 14的实验进一步证实DC通过可溶性CD 14依赖性途径对LPS应答。
To investigate the responses of dendritic cells (DC) during Gram-negative infections, we analyzed the effects of graded doses of LPS on the cytokine profile, phenotype, and allostimulatory potential of human DC generated by culturing plastic-adherent PBMC in presence of IL-4 and granulocyte-macrophage-CSF. First, we found that LPS stimulates the production of high levels of TNF-alpha, IL-6, IL-8, IL-12 by DC and up-regulates their expression of HLA-DR, B7-1, B7-2, and CD40. The effects of LPS were dose dependent, with a significant stimulatory effect already observed at a concentration of 0.1 ng/ml and a plateau being reached at 10 ng/ml. These phenotypic changes correlated with increased allostimulatory properties of LPS-activated DC because DC treated with LPS were significantly more efficient than untreated DC in eliciting IL-2 and IFN-gamma synthesis by alloreactive T cells and stimulating their proliferation. Experiments using neutralizing anti-IL-12 mAb indicated that LPS-induced IL-12 is responsible for the increased production of IFN-gamma but not for the increased proliferation during MLR. Finally, we observed that the DC responses to low levels of LPS (1 ng/ml) were dramatically inhibited by a blocking anti-CD14 mAb, although DC do not express CD14 molecules on their membrane. Experiments using serum depleted of soluble CD14 (sCD14) and sCD14 either purified from human serum or in recombinant form further established that DC respond to LPS via a soluble CD14-dependent pathway.