Pro-inflammatory cytokines and prostaglandins induce maturation of potent immunostimulatory dendritic cells under fetal calf serum-free conditions
Pro-inflammatory cytokines and prostaglandins induce maturation of potent immunostimulatory dendritic cells under fetal calf serum-free conditions
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DOI：
10.1002/eji.1830271209
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发表时间：
1997-12-01
影响因子：
5.4
通讯作者：
Enk, AH
中科院分区：
文献类型：
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作者：
Jonuleit, H;Kühn, U;Enk, AH
人类树突状细胞（DC）的培养条件已经由几个实验室开发出来。然而，大多数这些培养方法都是在存在粒细胞-巨噬细胞集落刺激因子和白细胞介素(IL)-4的情况下使用胎牛血清（FCS）来产生DC。最近，人们描述了使用单核细胞条件培养基（MCM）和无fcs培养基进行额外刺激以产生DC的替代培养条件。由于MCM是一种相当不确定的鸡尾酒，这些培养产生的DC的产量和质量差别很大。我们报道了肿瘤坏死因子(TNF)- α， IL-1 β和IL-6的确定混合物在其产生DC的效力上等于MCM。在细胞因子混合物中加入前列腺素（PG）E-2进一步提高了DC的产量、成熟、迁移和免疫刺激能力。更重要的是，培养条件也会影响诱导的T细胞反应的结果。单独使用tnf - α /IL-1/IL-6或MCM培养的DC诱导CD4(+) T细胞释放中等水平的干扰素(IFN)- γ，而不释放IL-4或IL-10。添加PGE可显著诱导ifn - γ的产生(2)，而对IL-4或IL-10的产生没有影响。在CD8(+) T细胞中观察到更显著的差异。MCM条件仅诱导ifn - γ（低），IL-4（负）细胞，tnf - α /IL-1/IL-6促进ifn - γ（中），IL-4（负）CD8(+) T细胞的生长。在没有诱导2型细胞因子的情况下，PGE(2)的加入只进一步极化了这一模式，增强了同种异体反应性CD8(+) T细胞产生ifn - γ的能力。综上所述，这些数据表明，定义的混合物tnf - α /IL-1/IL-6可以替代MCM， PGE(2)的加入进一步提高了DC的产量和质量。tnf - α /IL-1、IL-6 + PGE(2)培养的DC似乎最适合产生产生ifn - γ的CD4/CD8(+) T细胞。
Culture conditions for human dendritic cells (DC) have been developed by several laboratories. Most of these culture methods, however, have used conditions involving fetal calf serum (FCS) to generate DC in the presence of granulocyte-macrophage colony-stimulating factor and interleukin (IL)-4. Recently, alternative culture conditions have been described using an additional stimulation with monocyte-conditioned medium (MCM) and FCS-free media to generate DC. As MCM is a rather undefined cocktail, the yield and quality of DC generated by these cultures varies substantially. We report that a defined cocktail of tumor necrosis factor (TNF)-alpha, IL-1 beta and IL-6 equals MCM in its potency to generate DC. Addition of prostaglandin (PG)E-2 to the cytokine cocktail further enhanced the yield, maturation, migratory and immunostimulatory capacity of the DC generated. More importantly, culture conditions also influenced the outcome of the T cell response induced. DC cultured with TNF-alpha/IL-1/IL-6 or MCM alone induced CD4(+) T cells that release intermediate levels of interferon (IFN)-gamma and no IL-4 or IL-10. Production of IFN-gamma was significantly induced by addition of PGE(2), while no effect on production of IL-4 or IL-10 was observed. Even more striking differences were observed for CD8(+) T cells. While MCM conditions only induced IFN-gamma(low), IL-4(neg) cells, TNF-alpha/IL-1/IL-6 promoted growth of IFN-gamma(intermediate), IL-4(neg) CD8(+) T cells. Addition of PGE(2) again only further polarized this pattern enhancing IFN-gamma production by alloreactive CD8(+) T cells in both cultures without inducing type 2 cytokines. Taken together, the data indicate that the defined cocktail TNF-alpha/IL-1/IL-6 can substitute for MCM and that addition of PGE(2) further enhances the yield and quality of DC generated. TNF-alpha/IL-1, IL-6 + PGE(2)-cultured DC seem to be optimal for generation of IFN-gamma-producing CD4/CD8(+) T cells.