Molecular cloning and expression analysis of bovine programmed death-1
Molecular cloning and expression analysis of bovine programmed death-1
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DOI：
10.1111/j.1348-0421.2010.00208.x
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发表时间：
2010-05-01
影响因子：
2.6
通讯作者：
Ohashi, Kazuhiko
中科院分区：
文献类型：
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作者：
Ikebuchi, Ryoyo;Konnai, Satoru;Ohashi, Kazuhiko
最近的研究表明，PD-1是一种免疫抑制受体，参与肿瘤进展或慢性病毒感染期间下调免疫反应的机制。然而，在牛疾病的情况下，由于缺乏关于牛PD-1的信息，尚未有关于该分子的报道。在这项研究中，我们在两个品种的牛PD-1基因进行鉴定和初步表征。我们克隆了编码荷斯坦-弗里斯兰和日本黑牛PD-1的cDNA全序列，发现这两个基因都编码一个282个氨基酸的蛋白质，该蛋白质具有信号序列、跨膜结构域和基于免疫受体酪氨酸的抑制基序。这种牛PD-1与人和小鼠PD-1的同源性分别为72.9%和65.6%，两者均已得到充分表征和记录。定量实时PCR分析显示，牛PD-1主要在T细胞（如CD 4+和CD 8+细胞）和PBMC中表达，并且在T细胞刺激时通过ConA强烈上调。对有限数量的牛进行了检测，但正如预期的那样，患有牛白血病病毒诱导的淋巴瘤的牛中CD 4+和CD 8 + T细胞中PD-1 mRNA表达的程度高于未感染的牛。因此，需要进一步研究牛PD-1的功能，以阐明与牛中几种疾病和治疗进展相关的免疫抑制机制。
Recent work has shown that PD-1, an immune inhibitory receptor, is involved in mechanisms for down-regulating immune responses during tumor progression or chronic viral infection. However, in the case of bovine diseases, there have been no reports on this molecule due to lack of information about bovine PD-1. In this study, we performed identification and preliminary characterization of the bovine PD-1 gene in two breeds of cattle. We cloned full cDNA sequences encoding for PD-1 from both Holstein-Friesian and Japanese Black breeds, and found that both of the genes encoded a 282-amino acid protein, which had a signal sequence, transmembrane domain and an immunoreceptor tyrosine-based inhibitory motif. This bovine PD-1 showed 72.9% and 65.6% homology to human and mouse PD-1, respectively, both of which have been well characterized and documented. Quantitative real-time PCR analysis showed that bovine PD-1 is expressed predominantly in T-cells (such as CD4+ and CD8+ cells) and among PBMCs, and is strongly upregulated on T-cell stimulation via ConA. A limited number of cattle were tested yet, as expected, the degree of PD-1 mRNA expression in CD4+ and CD8+ T-cells was greater in cattle with bovine leukemia virus-induced lymphoma than in uninfected cattle. Further studies to characterize the functions of bovine PD-1 are therefore warranted, in order to elucidate the mechanism of the immunosuppression associated with progression of several diseases and therapy in cattle.