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目的分析ＤＲ９纯合细胞刺激下ＤＲ９阴性正常人ＴＣＲＢＶ基因片段的取用格局，确定和分离出ＤＲ９关联性疾病中的自身反应性Ｔ细胞克隆。方法常规方法获取ＰＢＭＣ经ＰＣＲ－ＳＳＯ和ＰＣＲ－ＲＦＬＰＤＮＡ分型，确定ＤＲ、ＤＱ、ＤＰ等位基因后，进行单向混合淋巴细胞培养，抽提总ＲＮＡ并合成第一链ｃＤＮＡ，定量ＰＣＲ检测２２种ＴＣＲＢＶ基因片段的取用格局。结果Ｔ细胞对ＤＲ９分子的识别和扩增具有寡克隆性，２例都有ＢＶ７和ＢＶ１６片段的优势取用，而片段ＢＶ１９和ＢＶ１０则在２例中分别表达。结论实验结果表明，共有ＢＶ７和ＢＶ１６片段的优势取用是ＤＲ９抗原等分子经直接途径激活了带有相应ＴＣＲＢＶ细胞的结果；而片段ＢＶ１９和ＢＶ１０在２例中分别表达可能是不同ＨＬＡ背景的ＡＰＣ间接递呈ＤＲ９抗原的结果。
OBJECTIVE: To analyze the access pattern of TCRBV gene fragment in DR9-negative normal subjects stimulated by DR9 homozygous cells and to identify and isolate autoreactive T cell clones in DR9-associated diseases. Methods PBMCs were genotyped by PCR-SSO and PCR-RFLP, and the DR, DQ and DP alleles were determined by routine methods. One-way mixed lymphocyte culture was performed. Total RNA was extracted and the first-strand cDNA was synthesized. TCRBV gene fragment access pattern. Results T cells showed oligoclonality for the recognition and amplification of DR9 molecules. Both of them had the advantage of using BV7 and BV16 fragments, while the fragments of BV19 and BV10 were respectively expressed in 2 cases. Conclusions The experimental results show that the advantages of shared BV7 and BV16 fragments are that the DR9 antigen and other molecules activate the corresponding TCRBV cells via direct pathway. The fragments BV19 and BV10 express APC Indirect presentation of DR9 antigen results.