摘要
目的 从单个细胞水平检测H R S细胞是否存在T细胞受体 (TCR) γ链基因重排 ,以研究其是否来源于分化早期的T淋巴细胞。方法 用显微操作仪从冷冻的新鲜活检组织切片中挑取CD30阳性的H R S细胞 ,提取其DNA并行聚合酶链反应 (PCR)。TCR γ基因重排的PCR采用半巢式扩增法 ,首轮用与TCR γ基因Vγ和Jγ区 (包括N区 )互补的引物对加入β 珠蛋白引物行混合扩增 ,对其中 β 珠蛋白扩增阳性的PCR产物进行第二轮PCR扩增 ,检测其TCR γ基因重排。 结果 在 6例霍奇金病冷冻切片中挑取的 5 6个CD30阳性的H R S细胞中有 2 4个细胞具有特异性的 2 6 8bp大小的β 珠蛋白PCR产物带。在第二轮扩增中 ,这 2 4个 β 珠蛋白阳性的H R S细胞均未发现有TCR γ基因重排。结论 所选病例中的H R S细胞并非来源于T淋巴细胞 ,可能来源于B淋巴细胞。
Objective To detect the rearrangement of T cell receptor γ chain gene in single H/R S cell as an indication of early T lymphocyte differentiation. Methods CD30 positive H/R S cells were isolated with micromanipulator from the frozen sections and analyzed by single cell PCR. TCR γ chain gene rearrangement was detected through semi nested PCR. In the first round of PCR, a pair of TCR γ primers which amplify the segments between Vγ and Jγ region (including the N region) of TCR γ gene mixing with a pair of β globin primers were used. In the second round, only the products positive β globin were amplified with the primers for TCR γ gene. Results A 268 bp β globin gene specific product was obtained in 24 out of 56 H/R S cells from 6 cases of Hodgkin′s disease. After the first round of PCR, no rearrangement of the TCR γ gene was noticed at a single cell level. Conclusion H/R S cells from the selected cases were not derived from the T cell lineage but possibly from the B cell lineage.
出处
《中华病理学杂志》
CAS
CSCD
北大核心
2000年第1期24-26,共3页
Chinese Journal of Pathology
基金
国家自然科学基金资助项目!(39470291)
关键词
何杰金氏病
H/R-S细胞
TCR-γ
基因重排
Hodgkin′s disease
Gene rearrangement, gamma-chain T-cell antigen receptor
Polymerase chain reaction
Micromanipulation
