霍奇金病单个H／R—S细胞TCR—γ基因重排的研究

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Ｐ

Analysis of TCR-gamma gene rearrangement in single H/R-S cells of Hodgkin disease

OBJECTIVE: To detect the rearrangement of T cell receptor gamma 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-gamma chain gene rearrangement was detected through semi-nested PCR. In the first round of PCR, a pair of TCR-gamma primers which amplify the segments between Vgamma and Jgamma region (including the N region) of TCR-gamma gene mixing with a pair of beta-globin primers were used. In the second round, only the products positive beta-globin were amplified with the primers for TCR-gamma gene. RESULTS: A 268 bp beta-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-gamma 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.