Functional analysis of bovine TLR5 and association with IgA responses of cattle following systemic immunisation with H7 flagella.
Functional analysis of bovine TLR5 and association with IgA responses of cattle following systemic immunisation with H7 flagella.
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DOI：
10.1186/s13567-014-0135-2
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发表时间：
2015-02-19
影响因子：
4.4
通讯作者：
Gally DL
中科院分区：
文献类型：
--
作者：
Tahoun A;Jensen K;Corripio-Miyar Y;McAteer SP;Corbishley A;Mahajan A;Brown H;Frew D;Aumeunier A;Smith DG;McNeilly TN;Glass EJ;Gally DL
鞭毛蛋白亚基是通过激活TLR 5（细胞外）和炎性小体（细胞内）来诱导宿主免疫应答的重要因子。我们以前的工作表明，用鞭毛系统免疫牛产生全身和粘膜伊加应答。小鼠中的伊加应答是TLR 5依赖性的，并且TLR 5可以影响适应性应答的总体幅度。然而，由于牛和人/鼠TLR 5序列之间的序列差异，尚不清楚牛TLR 5（bTLR 5）是否能够在与鞭毛蛋白相互作用后刺激炎症反应。为了解决这一问题，我们研究了含有bTLR 5的人和牛细胞对来自大肠杆菌的H7鞭毛蛋白的先天反应。大肠杆菌O 157：H7。与未转染的对照相比，用bTLR 5转染的HEK 293（人源）和胚胎牛肺（EBL）细胞均对H7鞭毛蛋白的添加作出响应。当突变引入H7鞭毛蛋白的TLR 5结合区时，包括R90 T取代，反应显著降低。在牛原代巨噬细胞中，当TLR 5转录水平被特异性siRNA抑制时，鞭毛蛋白刺激的CXCL 8 mRNA和分泌的蛋白水平显著降低，并且用R90 T-H7变体减少刺激。虽然这些结果表明bTLR 5序列产生功能性鞭毛蛋白识别受体，但与仅佐剂对照相比，用R90 T-H7鞭毛免疫的牛也表现出对鞭毛蛋白的全身性伊加应答。这可能反映了我们的发现，即R90 T-H7仍然激活bTLR 5，尽管与WT H7鞭毛蛋白相比效率降低，或者其他鞭毛蛋白识别途径可能在这种粘膜反应中起作用。
Flagellin subunits are important inducers of host immune responses through activation of TLR5 when extracellular and the inflammasome if cytosolic. Our previous work demonstrated that systemic immunization of cattle with flagella generates systemic and mucosal IgA responses. The IgA response in mice is TLR5-dependent and TLR5 can impact on the general magnitude of the adaptive response. However, due to sequence differences between bovine and human/murine TLR5 sequences, it is not clear whether bovine TLR5 (bTLR5) is able to stimulate an inflammatory response following interaction with flagellin. To address this we have examined the innate responses of both human and bovine cells containing bTLR5 to H7 flagellin from E. coli O157:H7. Both HEK293 (human origin) and embryonic bovine lung (EBL) cells transfected with bTLR5 responded to addition of H7 flagellin compared to non-transfected controls. Responses were significantly reduced when mutations were introduced into the TLR5-binding regions of H7 flagellin, including an R90T substitution. In bovine primary macrophages, flagellin-stimulated CXCL8 mRNA and secreted protein levels were significantly reduced when TLR5 transcript levels were suppressed by specific siRNAs and stimulation was reduced with the R90T-H7 variant. While these results indicate that the bTLR5 sequence produces a functional flagellin-recognition receptor, cattle immunized with R90T-H7 flagella also demonstrated systemic IgA responses to the flagellin in comparison to adjuvant only controls. This presumably either reflects our findings that R90T-H7 still activates bTLR5, albeit with reduced efficiency compared to WT H7 flagellin, or that other flagellin recognition pathways may play a role in this mucosal response.
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影响因子：
1.8
作者：
Metcalfe, Hannah J.;La Ragione, Roberto M.;Smith, David G. E.;Werling, Dirk
通讯作者：
Werling, Dirk
影响因子：
5.5
作者：
Farhat, Katja;Riekenberg, Sabine;Ulmer, Artur J.
通讯作者：
Ulmer, Artur J.
DOI：
10.1073/pnas.0502040102
发表时间：
2005-06-28
影响因子：
11.1
作者：
Andersen-Nissen, E;Smith, KD;Aderem, A
通讯作者：
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影响因子：
5.8
作者：
Lee, SK;Stack, A;Josenhans, C
通讯作者：
Josenhans, C
影响因子：
9.4
作者：
IBRAHIM, GF;FLEET, GH;WALKER, RA
通讯作者：
WALKER, RA