目的分析临床孤立综合征患者外周血T细胞受体β链可变区(TCRVβ)多态性,初步预测特异性TCRVβ片段,研究其对临床孤立综合征的诊断价值。方法选择2012年9月至2014年3月诊断并接受治疗的临床孤立综合征患者30例(CIS组),以同期行健康体检...目的分析临床孤立综合征患者外周血T细胞受体β链可变区(TCRVβ)多态性,初步预测特异性TCRVβ片段,研究其对临床孤立综合征的诊断价值。方法选择2012年9月至2014年3月诊断并接受治疗的临床孤立综合征患者30例(CIS组),以同期行健康体检的30例受试者为正常对照组。流式细胞术检测常规免疫学指标和TCRVβ亚家族表达变化,CELL Quest Pro 5.1等相关软件分析24个TCRVβ亚家族成员表达水平,并计算扩增者比例、扩增个数和扩增分数。结果TCRVβ24个亚家族中,CIS组CD4+TCRVβ2、4、5.2、5.3、7.1、8、14、17、20、21.3片段,以及CD8+TCRVβ4、14片段表达水平高于正常对照组,而CD4+TCRVβ11和CD8+TCRVβ9、11片段表达水平低于正常对照组(P<0.05或P<0.01)。CIS组CD4+TCRVβ(P=0.002)和CD8+TCRVβ14(P=0.010)扩增者比例均高于正常对照组,其余亚家族扩增者比例差异则无统计学意义(均P>0.05)。CIS组TCRVβ扩增者比例与正常对照组差异无统计学意义(P>0.05),但扩增个数(P=0.001)、扩增分数(P=0.006)均高于正常对照组。受试者工作特征(ROC)曲线显示,CD4+TCRVβ4诊断临床孤立综合征的曲线下面积(AUC)为0.719(95%CI:0.592~0.851,P=0.003),灵敏度83.33%、特异度56.67%;CD4+TCRVβ14诊断的AUC为0.713(95%CI:0.581~0.845,P=0.005),灵敏度70%、特异度56.23%;CD8+TCRVβ4诊断的AUC为0.727(95%CI:0.600~0.854,P=0.002),灵敏度93.33%、特异度50%;以上3个片段对临床孤立综合征均具有中等诊断价值。结论虽然临床孤立综合征患者外周血CD4+TCRVβ4、CD4+TCRVβ14和CD8+TCRVβ4片段表达水平升高,但其诊断价值有限。展开更多
AIM: To analyze the characterization of T-cell receptor-γ (TCR-γ) gene rearrangement in the gastrointestinal lymphomas and evaluate the value of PCR-SSCP analysis in gastrointestinal lymphomas investigation. METHODS...AIM: To analyze the characterization of T-cell receptor-γ (TCR-γ) gene rearrangement in the gastrointestinal lymphomas and evaluate the value of PCR-SSCP analysis in gastrointestinal lymphomas investigation. METHODS: TCR-γ gene rearrangement segments of gastrointestinal lymphomas were cloned and sequenced. Single clone plasmid and mixed clone plsamids were subsequently submitted to PCR-SSCP analysis to investigate the relationship between the number of amplified clones and band patterns of the amplified products. The PCR products of TCR-γ gene rearrangement of 40 gastrointestinal lymphomas were electrophoresed on agarose gels and the positive cases on agarose gels were studied by SSCP analysis. RESULTS: The sequencing showed that TCR-γ gene rearrangement of the gastrointestinal lyrnphomas included functional gene and pseudogene with extensive variety in the junctional regions. In SSCP analysis, the number of the single-stranded bands was about two times of the number of amplified clones, and double-stranded band became broad with the increased number of the amplified clones. Thirteen of the 25 B-cell gastrointestinal lymphornas and 14 of the 15 gastrointestinal T-cell lymphomas were positive detected on agarose gel electrophoresis. Of the positive cases detected by SSCP analysis, 3 B-cell lyrnphomas and 13 T-cell lyrnphomas showed positive bands. The other cases showed only smears. The rearranged pattern included 13 monoallelic gene rearrangements and 3 biallelic or oligoclonal gene rearrangements. CONCLUSION: The pattern of TCR-γ gene rearrangement in gastrointestinal lymphomas are similar to that of the nodular lymphomas. PCR-SSCP analysis for TCR-γ gene rearrangement can be applied both for adjuvant diagnosis of gastrointestinal lymphomas and analysis of the gene rearrangement pattern. The ratio of TCR-γ gene rearrangements occurred in T-cell gastrointestinal lymphomas is significantly higher than that in B-cell gastrointestinal lymphomas. The gene rearrangement pattern involves monoallelic and biallelic (or oligoclonal) gene rearrangement.展开更多
文摘目的分析临床孤立综合征患者外周血T细胞受体β链可变区(TCRVβ)多态性,初步预测特异性TCRVβ片段,研究其对临床孤立综合征的诊断价值。方法选择2012年9月至2014年3月诊断并接受治疗的临床孤立综合征患者30例(CIS组),以同期行健康体检的30例受试者为正常对照组。流式细胞术检测常规免疫学指标和TCRVβ亚家族表达变化,CELL Quest Pro 5.1等相关软件分析24个TCRVβ亚家族成员表达水平,并计算扩增者比例、扩增个数和扩增分数。结果TCRVβ24个亚家族中,CIS组CD4+TCRVβ2、4、5.2、5.3、7.1、8、14、17、20、21.3片段,以及CD8+TCRVβ4、14片段表达水平高于正常对照组,而CD4+TCRVβ11和CD8+TCRVβ9、11片段表达水平低于正常对照组(P<0.05或P<0.01)。CIS组CD4+TCRVβ(P=0.002)和CD8+TCRVβ14(P=0.010)扩增者比例均高于正常对照组,其余亚家族扩增者比例差异则无统计学意义(均P>0.05)。CIS组TCRVβ扩增者比例与正常对照组差异无统计学意义(P>0.05),但扩增个数(P=0.001)、扩增分数(P=0.006)均高于正常对照组。受试者工作特征(ROC)曲线显示,CD4+TCRVβ4诊断临床孤立综合征的曲线下面积(AUC)为0.719(95%CI:0.592~0.851,P=0.003),灵敏度83.33%、特异度56.67%;CD4+TCRVβ14诊断的AUC为0.713(95%CI:0.581~0.845,P=0.005),灵敏度70%、特异度56.23%;CD8+TCRVβ4诊断的AUC为0.727(95%CI:0.600~0.854,P=0.002),灵敏度93.33%、特异度50%;以上3个片段对临床孤立综合征均具有中等诊断价值。结论虽然临床孤立综合征患者外周血CD4+TCRVβ4、CD4+TCRVβ14和CD8+TCRVβ4片段表达水平升高,但其诊断价值有限。
基金Supported by Science and Technology Project of Guangzhou,No.2002Z3-E4061 Science and Technology Project of Guangdong,No.B30101
文摘AIM: To analyze the characterization of T-cell receptor-γ (TCR-γ) gene rearrangement in the gastrointestinal lymphomas and evaluate the value of PCR-SSCP analysis in gastrointestinal lymphomas investigation. METHODS: TCR-γ gene rearrangement segments of gastrointestinal lymphomas were cloned and sequenced. Single clone plasmid and mixed clone plsamids were subsequently submitted to PCR-SSCP analysis to investigate the relationship between the number of amplified clones and band patterns of the amplified products. The PCR products of TCR-γ gene rearrangement of 40 gastrointestinal lymphomas were electrophoresed on agarose gels and the positive cases on agarose gels were studied by SSCP analysis. RESULTS: The sequencing showed that TCR-γ gene rearrangement of the gastrointestinal lyrnphomas included functional gene and pseudogene with extensive variety in the junctional regions. In SSCP analysis, the number of the single-stranded bands was about two times of the number of amplified clones, and double-stranded band became broad with the increased number of the amplified clones. Thirteen of the 25 B-cell gastrointestinal lymphornas and 14 of the 15 gastrointestinal T-cell lymphomas were positive detected on agarose gel electrophoresis. Of the positive cases detected by SSCP analysis, 3 B-cell lyrnphomas and 13 T-cell lyrnphomas showed positive bands. The other cases showed only smears. The rearranged pattern included 13 monoallelic gene rearrangements and 3 biallelic or oligoclonal gene rearrangements. CONCLUSION: The pattern of TCR-γ gene rearrangement in gastrointestinal lymphomas are similar to that of the nodular lymphomas. PCR-SSCP analysis for TCR-γ gene rearrangement can be applied both for adjuvant diagnosis of gastrointestinal lymphomas and analysis of the gene rearrangement pattern. The ratio of TCR-γ gene rearrangements occurred in T-cell gastrointestinal lymphomas is significantly higher than that in B-cell gastrointestinal lymphomas. The gene rearrangement pattern involves monoallelic and biallelic (or oligoclonal) gene rearrangement.
