Transcriptomic profiling of bovine blood dendritic cells and monocytes following TLR stimulation.,European Journal of Immunology

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Transcriptomic profiling of bovine blood dendritic cells and monocytes following TLR stimulation.
European Journal of Immunology ( IF 4.5 ) Pub Date : 2020-06-27 , DOI: 10.1002/eji.202048643
G Tuba Barut _{1,

2,

3} , Heidi E L Lischer ₄ , Rémy Bruggmann ₄ , Artur Summerfield _{1,

2} , Stephanie C Talker _{1,

2}

Affiliation

Dendritic cells (DC) and monocytes are vital for the initiation of innate and adaptive immune responses. Recently, we identified bona fide DC subsets in blood of cattle, revealing subset‐ and species‐specific transcription of toll‐like receptors (TLR). In the present study, we analyzed phenotypic and transcriptional responses of bovine DC subsets and monocytes to in vitro stimulation with four to six different TLR ligands. Bovine DC subsets, especially plasmacytoid DC (pDC), showed a clear increase of CCR7, CD25, CD40, CD80, CD86, and MHC‐II expression both on mRNA and protein level. Flow cytometric detection of p38 MAPK phosphorylation 15 min after stimulation confirmed activation of DC subsets and monocytes in accordance with TLR gene expression. Whole‐transcriptome sequencing of sorted and TLR‐stimulated subsets revealed potential ligand‐ and subset‐specific regulation of genes associated with inflammation, T‐cell co‐stimulation, migration, metabolic reprogramming, and antiviral activity. Gardiquimod was found to evoke strong responses both in DC subsets and monocytes, while Poly(I:C) and CpG preferentially triggered responses in cDC1 and pDC, respectively. This in‐depth analysis of ligand responsiveness is essential for the rational design of vaccine adjuvants in cattle, and provides a solid basis for comparative studies on DC and monocyte biology across species.

中文翻译：

TLR刺激后牛血树突状细胞和单核细胞的转录组分析。

树突状细胞（DC）和单核细胞对于启动先天性和适应性免疫反应至关重要。最近，我们确定了善意牛血液中的DC亚群，揭示了Toll样受体（TLR）的亚群和物种特异性转录。在本研究中，我们分析了牛DC亚群和单核细胞对体外刺激4-6种不同TLR配体的表型和转录反应。牛DC亚群，尤其是浆细胞样DC（pDC）在mRNA和蛋白质水平上均显示CCR7，CD25，CD40，CD80，CD86和MHC-II表达明显增加。刺激后15分钟，流式细胞术检测p38 MAPK磷酸化，证实DC子集和单核细胞根据TLR基因表达被激活。排序和TLR刺激的子集的全转录组测序揭示了与炎症，T细胞共刺激，迁移，代谢重编程，和抗病毒活性。Gardiquimod被发现在DC亚群和单核细胞中均引起强烈反应，而Poly（I：C）和CpG分别在cDC1和pDC中优先引发反应。这种对配体反应性的深入分析对于牛类疫苗佐剂的合理设计至关重要，并且为跨物种的DC和单核细胞生物学的比较研究提供了坚实的基础。

更新日期：2020-06-27

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