Use of murine embryonic fibroblasts to define Toll-like receptor activation and specificity
Use of murine embryonic fibroblasts to define Toll-like receptor activation and specificity
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DOI：
10.1179/096805104225006516
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发表时间：
2004-01-01
期刊：
影响因子：
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通讯作者：
Finberg, RW
中科院分区：
文献类型：
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作者：
Kurt-Jones, EA;Sandor, F;Finberg, RW
Toll样受体(TLRs)在细菌、病毒和真菌病原体的先天免疫反应中起重要作用。我们研究了人外周血单个核细胞、小鼠胚胎成纤维细胞(MEF)和一组人细胞系，包括HEK、HeLa、AGS、ECV304和U373细胞，以观察TLR特异性mRNAs的表达和TLR配体依赖的细胞因子的分泌。外周血细胞表达多种TLR；然而，许多研究表明，血液中含有多种不同的细胞群体，具有不同的TLR表达模式。TLR2、TLR8表达缺失，TLR5、TLR7、TLR9表达较弱。相反，MEF高表达TLR1、TLR2、TLR3、TLR4、TLR5、TLR6、TLR7、TLR8和TLR9的mRNA。MEF对TLR配体激活高度敏感，对TLR配体反应分泌高水平的IL-6和MCP-1。来自TLR2、TLR4和MyD88靶向缺失的小鼠的MEF显示，它们对特定配体的IL-6反应存在严重缺陷，这与对成年基因敲除动物的巨噬细胞和组织的研究一致。MEF培养是同质的，易于进行生化分析，并应允许对单个TLR对先天性免疫反应的贡献进行严格的研究。
Toll-like receptors (TLRs) are critically involved in the innate immune response to bacterial, viral and fungal pathogens. We have studied human peripheral blood mononuclear cells, murine embryonic fibroblasts (MEFs) and a panel of human cell lines, including HEK, HeLa, AGS, ECV304 and U373 cells, for expression of TLR-specific mRNAs and for TLR-ligand dependent cytokine secretion. Peripheral blood cells expressed multiple TLRs; however, many studies have shown that blood contains multiple, heterogeneous cell populations with distinct patterns of TLR expression. Cell lines had variable expression of TLRs, and in most cases lacked TLR2 and TLR8 expression and only weakly expressed mRNAs for TLR5, TLR7 and TLR9. In contrast, MEFs expressed high levels of mRNA for TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8 and TLR9. MEFs were highly responsive to TLR-ligand activation and secreted high levels of both IL-6 and MCP-1 in response to TLR ligands. MEFs from mice with targeted deletions of TLR2, TLR4 and MyD88 demonstrated profound defects in their IL-6 response to their specific ligands, consistent with studies of macrophages and tissues from adult knockout animals. MEF cultures are homogenous and amenable to biochemical analysis and should allow rigorous studies of the contribution of individual TLRs to the innate immune response.