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目的初步分析1月龄Balb/c小鼠胸腺、脾脏、肝脏、小肠和血液中TRBV-TRBJ1-1 CDR3谱型特征。方法提取1月龄Balb/c小鼠胸腺、脾脏、肝脏、小肠和血液样本的总RNA,逆转录成cDNA,以22条TRBV基因家族为上游引物,共同的TRBJ1-1基因为下游引物(荧光标记),PCR扩增包含完整CDR3区片断,毛细管电泳激光DNA扫描技术分析CDR3谱型的长度和多态性。结果胸腺样本毛细管电泳激光扫描图呈现标准的中间高两端低的多态性分布;各家族CDR3区最长和最短比较,多数为相差8～15个氨基酸的谱型分布。脾脏、肝脏、小肠、血液样本扫描图显示多数家族呈多态性分布,但均出现数量不等的单峰、寡峰、偏峰、低峰;多数家族表现为相差3～13个氨基酸的谱型分布,部分家族在预测大小位置无对应峰型。结论 1月龄Balb/c小鼠胸腺中基本包含TCR基因随机重排的多种克隆性T细胞;脾脏、肝脏、小肠、血液表现为单、寡、多克隆形式的T细胞分布,部分家族T细胞缺失,本实验结果可为进一步研究Balb/c小鼠的T细胞发育、分布、应答及CDR3谱型与疾病关系提供方法和基础。
OBJECTIVE: To characterize the CDR3 pattern of TRBV-TRBJ1-1 in the thymus, spleen, liver, small intestine and blood of 1-month-old Balb / c mice. Methods Total RNA was extracted from the thymus, spleen, liver, small intestine and blood samples of 1-month-old Balb / c mice and reverse transcribed into cDNA. Twenty-two TRBV genes were used as upstream primers and common TRBJ1-1 genes as downstream primers Marker), PCR amplification of the entire CDR3 region containing fragments, capillary electrophoresis laser DNA scanning technology to analyze CDR3 profile length and polymorphism. Results The capillary scanning electrophoresis of thymus samples showed a low polymorphism distribution at the middle and the high end of each sample. The longest and shortest CDR3 regions in most families were found, most of which were 8-15 amino acids in spectral distribution. The spleen, liver, small intestine and blood samples showed most of the families were polymorphic, but there were a number of single peak, low peak, partial peak and low peak; most families showed a spectrum of 3 to 13 amino acids Type distribution, some families in the predicted size location no corresponding peak. CONCLUSIONS: Many clonogenic T cells in the thymus of 1-month-old Balb / c mice are randomly rearranged with TCR genes. The spleen, liver, small intestine and blood show the distribution of T cells in single, oligo and polyclonal forms. Some T The results of this study provide a method and basis for further study of T cell development, distribution and response of Balb / c mice and the relationship between CDR3 patterns and diseases.