TNF-α augments RANKL-dependent intestinal M cell differentiation in enteroid cultures.
TNF-α augments RANKL-dependent intestinal M cell differentiation in enteroid cultures.
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TNF-α增强了肠培养基中RANKL依赖性肠细胞分化。
DOI：
10.1152/ajpcell.00108.2016
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发表时间：
2016-09-01
期刊：
影响因子：
--
通讯作者：
Williams IR
中科院分区：
文献类型：
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作者：
Wood MB;Rios D;Williams IR
微褶（M）细胞是Peyer集合淋巴结滤泡相关上皮中的吞噬性肠上皮细胞，可将颗粒抗原从肠腔转运至上皮下圆顶。肠隐窝上皮干细胞分化为M细胞需要NF-κB配体的细胞因子受体激活因子（RANKL）和转录因子Spi-B。我们使用小鼠小肠隐窝建立的三维肠样培养物作为模型系统，以研究参与M细胞分化的信号转导途径以及其他细胞因子对RANKL诱导的M细胞分化的影响。向类肠细胞中添加RANKL可诱导多个M细胞相关基因的表达，包括Spib、Ccl 9 [趋化因子（C-C基序）配体9]、Tnfaip 2（TNF-α诱导蛋白2）、Anxa 5（膜联蛋白A5）和Marcksl 1（肉豆蔻酰化富含丙氨酸的蛋白激酶C底物）。成熟的M细胞标志物糖蛋白2（Gp 2）被强烈诱导3天，并在11%的细胞在肠样组织中表达。非经典NF-κB途径是类肠细胞中RANKL诱导的M细胞分化所必需的，因为向编码NF-κ B诱导激酶的丝裂原活化蛋白激酶14（Map 3 k14）基因无效突变小鼠的类肠细胞中添加RANKL未能诱导M细胞相关基因。虽然单独使用细胞因子TNF-α对M细胞相关基因的表达几乎没有影响（如果有的话），但在RANKL中添加TNF-α始终导致多种M细胞相关基因的水平比单独使用RANKL高3 - 6倍。一种作用机制是TNF-α快速诱导RelB和Nfkb 2（NF-κB亚基2），这两种基因编码非经典NF-κB异源二聚体的两个亚基。我们得出结论，肠相关淋巴组织微环境中存在的经典NF-κB信号传导的内源性激活剂（包括TNF-α）可在滤泡相关上皮中M细胞的RANKL依赖性分化中发挥支持作用。
Microfold (M) cells are phagocytic intestinal epithelial cells in the follicle-associated epithelium of Peyer's patches that transport particulate antigens from the gut lumen into the subepithelial dome. Differentiation of M cells from epithelial stem cells in intestinal crypts requires the cytokine receptor activator of NF-κB ligand (RANKL) and the transcription factor Spi-B. We used three-dimensional enteroid cultures established with small intestinal crypts from mice as a model system to investigate signaling pathways involved in M cell differentiation and the influence of other cytokines on RANKL-induced M cell differentiation. Addition of RANKL to enteroids induced expression of multiple M cell-associated genes, including Spib, Ccl9 [chemokine (C-C motif) ligand 9], Tnfaip2 (TNF-α-induced protein 2), Anxa5 (annexin A5), and Marcksl1 (myristoylated alanine-rich protein kinase C substrate) in 1 day. The mature M cell marker glycoprotein 2 (Gp2) was strongly induced by 3 days and expressed by 11% of cells in enteroids. The noncanonical NF-κB pathway was required for RANKL-induced M cell differentiation in enteroids, as addition of RANKL to enteroids from mice with a null mutation in the mitogen-activated protein kinase kinase kinase 14 (Map3k14) gene encoding NF-κB-inducing kinase failed to induce M cell-associated genes. While the cytokine TNF-α alone had little, if any, effect on expression of M cell-associated genes, addition of TNF-α to RANKL consistently resulted in three- to sixfold higher levels of multiple M cell-associated genes than RANKL alone. One contributing mechanism is the rapid induction by TNF-α of Relb and Nfkb2 (NF-κB subunit 2), genes encoding the two subunits of the noncanonical NF-κB heterodimer. We conclude that endogenous activators of canonical NF-κB signaling present in the gut-associated lymphoid tissue microenvironment, including TNF-α, can play a supportive role in the RANKL-dependent differentiation of M cells in the follicle-associated epithelium.