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A comprehensive analysis of TDO2 expression in immune cells and characterization of immune cell phenotype in TDO2 knockout mice
Transgenic Research ( IF 2.7 ) Pub Date : 2021-09-16 , DOI: 10.1007/s11248-021-00281-8
Susu Li 1 , Siyu Li 1 , Yingjie Zhao 1 , Bingjie Zhang 1 , Xinwei Wang 1 , Xuezhi Yang 1 , Yueye Wang 1 , Chengyan Jia 1 , Yan Chang 1 , Wei Wei 1
Affiliation  

Tryptophan 2,3-dioxygenase (TDO2) was an initial rate-limiting enzyme of the kynurenine (Kyn) pathway in tryptophan (Trp) metabolism. We undertook this study to determine a comprehensive analysis of TDO2 expression in immune cells and assess the characterization of immune cell phenotype in TDO2 knockout mice. The expression of TDO2 in various tissues of DBA/1 mice was detected by quantitative real-time PCR (qPCR) and immunohistochemistry. Both flow cytometry and immunofluorescence were used to analyze the expression of TDO2 in immune cells. Furthermore, TDO2 knockout (KO) mice were generated by CRISPR/Cas9 technology to detect immune cell phenotype. TDO2 protein level in liver was tested by western blot. High-performance liquid chromatography was used to detect the level of Trp and Kyn. Flow cytometry was used to test the proportions of splenic lymphocyte subsets in wild-type (WT) and TDO2 KO mice. We found that TDO2 was expressed in various tissues and immune cells, and TDO2 staining was mainly observed in the cytoplasm of cells. There was no difference in the development of immune cells between TDO2 KO mice and WT mice, including T cells, B cells, memory B cells, plasma cells, dendritic cells, and natural killer cells. Interestingly, the reduced M1/M2 ratio was observed in the peritoneal macrophages of TDO2 KO mice. Taken together, these findings enriched the known expression profile of TDO2, especially its expression in immune cells. Our study suggested that TDO2-mediated Trp-Kyn metabolism pathway might be involved in the immune response.



中文翻译:

TDO2基因敲除小鼠免疫细胞中TDO2表达及免疫细胞表型特征的综合分析

色氨酸 2,3-双加氧酶 (TDO2) 是色氨酸 (Trp) 代谢中犬尿氨酸 (Kyn) 途径的初始限速酶。我们进行了这项研究,以确定免疫细胞中 TDO2 表达的综合分析,并评估 TDO2 敲除小鼠中免疫细胞表型的表征。采用定量实时PCR(qPCR)和免疫组化方法检测DBA/1小鼠不同组织中TDO2的表达。流式细胞术和免疫荧光均用于分析TDO2在免疫细胞中的表达。此外,通过 CRISPR/Cas9 技术生成 TDO2 敲除 (KO) 小鼠以检测免疫细胞表型。通过蛋白质印迹测试肝脏中的TDO2蛋白水平。使用高效液相色谱法检测Trp和Kyn的水平。流式细胞仪用于检测野生型 (WT) 和 TDO2 KO 小鼠脾淋巴细胞亚群的比例。我们发现TDO2在各种组织和免疫细胞中均有表达,TDO2染色主要在细胞的细胞质中观察到。TDO2 KO小鼠和WT小鼠的免疫细胞发育没有差异,包括T细胞、B细胞、记忆B细胞、浆细胞、树突状细胞和自然杀伤细胞。有趣的是,在 TDO2 KO 小鼠的腹腔巨噬细胞中观察到 M1/M2 比率降低。总之,这些发现丰富了 TDO2 的已知表达谱,尤其是它在免疫细胞中的表达。我们的研究表明 TDO2 介导的 Trp-Kyn 代谢途径可能参与免疫反应。我们发现TDO2在各种组织和免疫细胞中均有表达,TDO2染色主要在细胞的细胞质中观察到。TDO2 KO小鼠和WT小鼠的免疫细胞发育没有差异,包括T细胞、B细胞、记忆B细胞、浆细胞、树突状细胞和自然杀伤细胞。有趣的是,在 TDO2 KO 小鼠的腹腔巨噬细胞中观察到 M1/M2 比率降低。总之,这些发现丰富了 TDO2 的已知表达谱,尤其是它在免疫细胞中的表达。我们的研究表明 TDO2 介导的 Trp-Kyn 代谢途径可能参与免疫反应。我们发现TDO2在各种组织和免疫细胞中均有表达,TDO2染色主要在细胞的细胞质中观察到。TDO2 KO小鼠和WT小鼠的免疫细胞发育没有差异,包括T细胞、B细胞、记忆B细胞、浆细胞、树突状细胞和自然杀伤细胞。有趣的是,在 TDO2 KO 小鼠的腹腔巨噬细胞中观察到 M1/M2 比率降低。总之,这些发现丰富了 TDO2 的已知表达谱,尤其是它在免疫细胞中的表达。我们的研究表明 TDO2 介导的 Trp-Kyn 代谢途径可能参与免疫反应。包括T细胞、B细胞、记忆B细胞、浆细胞、树突细胞和自然杀伤细胞。有趣的是,在 TDO2 KO 小鼠的腹腔巨噬细胞中观察到 M1/M2 比率降低。总之,这些发现丰富了 TDO2 的已知表达谱,尤其是它在免疫细胞中的表达。我们的研究表明 TDO2 介导的 Trp-Kyn 代谢途径可能参与免疫反应。包括T细胞、B细胞、记忆B细胞、浆细胞、树突细胞和自然杀伤细胞。有趣的是,在 TDO2 KO 小鼠的腹腔巨噬细胞中观察到 M1/M2 比率降低。总之,这些发现丰富了 TDO2 的已知表达谱,尤其是它在免疫细胞中的表达。我们的研究表明 TDO2 介导的 Trp-Kyn 代谢途径可能参与免疫反应。

更新日期:2021-09-16
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