CD3γ/δ in sea bass (Dicentrarchus labrax): Molecular characterization and expression analysis

Results in immunology Pub Date : 2011-05-01 Epub Date: 2011-08-30 DOI:10.1016/j.rinim.2011.08.003
Elisa Randelli, Giuseppe Scapigliati, Francesco Buonocore
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引用次数: 9

Abstract

The CD3 complex is the common marker on the surface of both αβ and γδ T cells and is essential for formation of the T-cell receptor complex and for T-cell activation.

In this paper, we report the gene cloning and molecular characterization of a CD3γ/δ homologue in sea bass (Dicentrarchus labrax), the analysis of transcription levels in lymphoid and non-lymphoid organs and the gene regulation after in vitro stimulation with LPS and PHA.

Four cysteine residues in the extracellular domain, involved in the constitution of immunoglobulin-like domain, are present in sea bass CD3γ/δ sequence and they are conserved both in number and position from mammals to teleost sequences. Similar to other known CD3γ/δs, in sea bass CD3γ/δ there is also a conserved immunoreceptor tyrosine-based activation ITAM motif that could be responsible for its individual signal transduction capacity.

The real time RT-PCR basal analysis shows the highest level of CD3γ/δ mRNA in thymus, followed by peripheral blood leucocytes, spleen, gills, gut, liver, head kidney, brain and muscle. The expression analysis under stimuli condition reveals a significant decrease of CD3γ/δ expression after LPS stimulation and a significant increase after PHA-L stimulation, in agreement with mammals results.

In conclusion, these data allow us to affirm that sea bass CD3γ/δ can be used as a T cell marker and will help in adding new insight on the immune response mechanisms of sea bass.

黑鲈(Dicentrarchus labrax) CD3γ/δ的分子特征及表达分析
CD3复合物是αβ和γδ T细胞表面的共同标记物,对T细胞受体复合物的形成和T细胞活化至关重要。本文报道了海鲈鱼(Dicentrarchus labrax)中CD3γ/δ同源基因的克隆和分子特征,分析了其在淋巴和非淋巴器官中的转录水平以及LPS和PHA体外刺激后的基因调控。在黑鲈CD3γ/δ序列中,存在4个参与免疫球蛋白样结构域构成的胞外结构域半胱氨酸残基,从哺乳动物到硬骨鱼,它们的数量和位置都是保守的。与其他已知的CD3γ/δs类似,在黑鲈CD3γ/δ中也存在一个保守的基于酪氨酸的免疫受体激活ITAM基序,该基序可能负责其个体信号转导能力。实时RT-PCR基础分析显示,胸腺中CD3γ/δ mRNA水平最高,其次是外周血白细胞、脾脏、鳃、肠道、肝脏、头肾、脑和肌肉。刺激条件下的表达分析显示,LPS刺激后CD3γ/δ表达显著降低,PHA-L刺激后CD3γ/δ表达显著升高,与哺乳动物实验结果一致。综上所述,这些数据使我们确认了黑鲈CD3γ/δ可以作为T细胞标记物,并将有助于对黑鲈免疫反应机制的新认识。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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