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欧洲鲈鱼发育过程中 TRγ 链的分子、细胞和功能分析。

Molecular, Cellular and Functional Analysis of TRγ Chain along the European Sea Bass Development.

机构信息

Department for Innovation in Biological, Agro-Food and Forest Systems, University of Tuscia, Largo dell'Università, 01100 Viterbo, Italy.

Section of Electron Microscopy, Great Equipment Center, University of Tuscia, 01100 Viterbo, Italy.

出版信息

Int J Mol Sci. 2021 Mar 25;22(7):3376. doi: 10.3390/ijms22073376.

Abstract

In jawed vertebrates, adaptive immune responses are enabled by T cells. Two lineages were characterized based on their T cell receptor (TcR) heterodimers, namely αβ or γδ peptide chains, which display an Ig domain-type sequence that is somatically rearranged. γδ T cells have been less extensively characterized than αβ and teleost fish, in particular, suffer from a severe scarcity of data. In this paper, we worked on the well-known model, the European sea bass to broaden the understanding of teleost γδ-T cells. The T cell receptor chain (TR) γ transcript was expressed at a later developmental stage than TRβ, suggesting a layered appearance of fish immune cells, and the thymus displayed statistically-significant higher mRNA levels than any other organ or lymphoid tissue investigated. The polyclonal antibody developed against the TRγ allowed the localization of TRγ-expressing cells in lymphoid organs along the ontogeny. Cell positivity was investigated through flow cytometry and the highest percentage was found in peripheral blood leukocytes, followed by thymus, gut, gills, spleen and head kidney. Numerous TRγ-expressing cells were localized in the gut mucosa, and the immunogold labelling revealed ultrastructural features that are typical of T cells. At last, microalgae-based diet formulations significantly modulated the abundance of TRγ cells in the posterior intestine, hinting at a putative involvement in nutritional immunity. From a comparative immunological perspective, our results contribute to the comprehension of the diversity and functionalities of γδ T cells during the development of a commercially relevant marine teleost model.

摘要

在有颌脊椎动物中,适应性免疫反应是由 T 细胞启动的。根据 T 细胞受体(TcR)异二聚体,即 αβ 或 γδ 肽链的特征,可以将其分为两个谱系,这些肽链显示出具有体细胞重排的 Ig 结构域类型序列。γδ T 细胞的特征不如 αβ T 细胞广泛,尤其是在硬骨鱼类中,数据非常匮乏。在本文中,我们以著名的欧洲鲈鱼模型为研究对象,旨在拓宽对硬骨鱼 γδ-T 细胞的理解。T 细胞受体链(TR)γ的转录本在比 TRβ 更晚的发育阶段表达,这表明鱼类免疫细胞的出现具有分层性,并且胸腺的 mRNA 水平比研究的任何其他器官或淋巴组织都高,具有统计学意义。针对 TRγ 开发的多克隆抗体允许在沿个体发生过程中的淋巴器官中定位表达 TRγ 的细胞。通过流式细胞术研究细胞阳性率,发现外周血白细胞中的阳性率最高,其次是胸腺、肠道、鳃、脾脏和头肾。在肠道黏膜中定位了大量表达 TRγ 的细胞,免疫金标记揭示了 T 细胞的典型超微结构特征。最后,基于微藻的饮食配方显著调节了后肠中 TRγ 细胞的丰度,提示其可能参与营养免疫。从比较免疫学的角度来看,我们的研究结果有助于理解商业相关海洋硬骨鱼类模型中 γδ T 细胞的多样性和功能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3ab1/8036326/dff95f673cd8/ijms-22-03376-g001.jpg

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