Hodges E, Edwards S E, Howell W M, Smith J L
Molecular Immunology Group, Southampton University Hospitals NHS Trust, UK.
Leukemia. 1994 Feb;8(2):295-8.
Clonality in T-cell malignancy was investigated using T-cell receptor (TcR) V beta 1-20 family primers and polymerase chain reaction amplification (PCR) of cDNA prepared from tissue biopsies and blood involved with tumour. Secondary PCR amplification of the VDJ joints of primary PCR products was performed to distinguish clonal from polyclonal products, and clonal V beta gene products were confirmed by direct PCR sequencing in the majority of cases. In eight T-cell malignancies including T-cell acute lymphoblastic leukaemia (T-ALL) and T-cell chronic lymphocytic leukaemia (T-CLL) shown to be clonal by Southern blot analysis, one or two primary PCR products were identified and shown to be clonal. In five cases of peripheral T-cell lymphoma (PTCL) all V beta 1-20 families were identified after primary PCR amplification, and clonal products were identified in two cases after secondary amplification; TcR V beta clonal families could not be demonstrated in the remaining three cases. These data were in agreement with previous Southern blot analysis of these cases, and confirmed the presence of reactive T cells in PTCL as well as providing further evidence for the genotypic heterogeneity of this entity. In the remaining case, a blood lymphocytosis, primary PCR amplification produced predominant TcR V beta 6 and V beta 12 family products, of which the V beta 6 family proved clonal after secondary PCR amplification. There was no evidence for overrepresentation of TCR V beta families by the tumour populations in this study, furthermore the data confirm the involvement of reactive cells in T-cell malignancy and the genetic heterogeneity of PTCL.
利用T细胞受体（TcR）Vβ1 - 20家族引物以及从肿瘤相关组织活检和血液中制备的cDNA进行聚合酶链反应扩增（PCR），研究T细胞恶性肿瘤中的克隆性。对一级PCR产物的VDJ接头进行二级PCR扩增，以区分克隆产物和多克隆产物，并且在大多数情况下通过直接PCR测序确认克隆性Vβ基因产物。在通过Southern印迹分析显示为克隆性的8例T细胞恶性肿瘤中，包括T细胞急性淋巴细胞白血病（T - ALL）和T细胞慢性淋巴细胞白血病（T - CLL），鉴定出一或两个一级PCR产物并显示为克隆性。在5例外周T细胞淋巴瘤（PTCL）中，一级PCR扩增后鉴定出所有Vβ1 - 20家族，二级扩增后在2例中鉴定出克隆产物；在其余3例中未证实TcR Vβ克隆家族。这些数据与之前对这些病例的Southern印迹分析一致，证实了PTCL中反应性T细胞的存在，并为该实体的基因型异质性提供了进一步证据。在其余病例中，一名血液淋巴细胞增多症患者，一级PCR扩增产生主要的TcR Vβ6和Vβ12家族产物，其中Vβ6家族在二级PCR扩增后被证明是克隆性的。本研究中没有证据表明肿瘤群体中TCR Vβ家族过度表达，此外，数据证实反应性细胞参与T细胞恶性肿瘤以及PTCL的基因异质性。
