Zambello R, Trentin L, Facco M, Cerutti A, Sancetta R, Milani A, Raimondi R, Tassinari C, Agostini C, Semenzato G
Department of Clinical and Expedrimental Medicine, Padua University School of Medicine, Italy.
Cancer Res. 1995 Dec 15;55(24):6140-5.
To investigate whether cell populations in CD3+ lymphoproliferative disease of granular lymphocytes (LDGLs) were skewed toward the use of specific V beta regions, we studied the repertoire of T-cell receptor (TCR) V beta gene products in 18 patients, as well as their relationship to the clonal bands in the Southern blot and the activation mediated by superantigens. Using a panel of monoclonal antibodies (mAbs) for conserved V beta segments and PCR, a dominant population expressing a specific V beta region was demonstrated in all patients. In five (27%) cases, granular lymphocytes (GLs) were found to express the V beta 13.1, while V beta 8 and V beta 6 were each expressed in three (17%) cases. The remaining cases were characterized by the proliferation of TCR V beta 2, V beta 3, V beta 4, V beta 9, V beta 12, V beta 16, and V beta 20. This finding indicates a biased usage of a limited TCR V beta in LDGLs, since nearly 60% of the cases utilized only three families of the TCR V beta genes. In all of the cases studied, we proved that the subset recognized by mAb and PCR was identical to that accounting for the extra band(s) of the Southern blot. This finding confirms the clonal nature of the population identified according to TCR V beta expression both by phenotype and PCR. On functional grounds, we evaluated whether GLs can be activated through the specific TCR using the superantigens recognizing discrete V beta families, such as staphylococcal proteins, including SEA, SEB, SEC1, SEC2, SED, and SEE. We demonstrated that the TCR-alpha/beta of clonal GLs in LDGL patients is functionally active in delivering cytotoxic and proliferative signals upon superantigen activation.
为了研究颗粒淋巴细胞（GLs）的CD3⁺淋巴细胞增殖性疾病（LDGLs）中的细胞群体是否偏向于使用特定的Vβ区域，我们研究了18例患者的T细胞受体（TCR）Vβ基因产物库，以及它们与Southern印迹中的克隆条带和超抗原介导的激活之间的关系。使用一组针对保守Vβ片段的单克隆抗体（mAb）和PCR，在所有患者中均证实了表达特定Vβ区域的优势群体。在5例（27%）病例中，发现颗粒淋巴细胞（GLs）表达Vβ13.1，而Vβ8和Vβ6在3例（17%）病例中各有表达。其余病例的特征是TCR Vβ2、Vβ3、Vβ4、Vβ9、Vβ12、Vβ16和Vβ20增殖。这一发现表明LDGLs中TCR Vβ的使用存在偏向性，因为近60%的病例仅使用了三个TCR Vβ基因家族。在所有研究的病例中，我们证明了mAb和PCR识别的亚群与Southern印迹中额外条带所对应的亚群相同。这一发现证实了根据TCR Vβ表达通过表型和PCR鉴定的群体的克隆性质。基于功能方面的考虑，我们使用识别离散Vβ家族的超抗原，如包括SEA、SEB、SEC1、SEC2、SED和SEE在内的葡萄球菌蛋白，评估GLs是否可以通过特异性TCR被激活。我们证明，LDGL患者克隆性GLs的TCR-α/β在超抗原激活后在传递细胞毒性和增殖信号方面具有功能活性。
