Schwab Cornelia, Willers Jörg, Niederer Eva, Ludwig Elisabeth, Kündig Thomas, Grob Peter, Burg Günter, Dummer Reinhard
Department of Dermatology, University Hospital of Zurich, Zurich, Switzerland.
Br J Haematol. 2002 Sep;118(4):1019-26. doi: 10.1046/j.1365-2141.2002.03726.x.
Sézary syndrome and Mycosis fungoides are the most common forms of cutaneous T-cell lymphomas. To assess the response to different therapies especially in Sézary syndrome, it is helpful to monitor the percentage of circulating tumour cells in the blood. The use of T-cell receptor (TCR)-Vbeta specific monoclonal antibodies provides a suitable tool for detecting Sézary cells. In this study, we analysed the levels of clonal CD4+Vbeta+ cells of seven patients with various treatment modalities using flow cytometry and investigated the immunophenotype of the clonal cells by double staining with a panel of antibodies recognizing lymphatic surface markers. Additionally, a polymerase chain reaction-denaturing gradient gel electrophoresis assay was performed on clonal CD4+Vbeta2+ cells, showing that these cells carry a Vgamma10/11, JgammaP1/2 TCR rearrangement. Follow-up studies revealed close association of the Vbeta+ clone developmentwith the clinical response to different therapiesinsixpatients. Intwo cases, the CD4+Vbeta+ cells decreased accompanied by partial regression or even complete remission. In four cases, a stable or increasing clonal CD4+Vbeta+ population reflected well a stable or progressing course of the disease. Double staining of Vbeta+ cells revealed the following pattern, CD3+, CD5+, CD7+, CD28+, CD80-, CD86+ and human leucocyte antigen (HLA) class I+. In contrast, HLA-DR was heterogeneously expressed. We conclude that identification and monitoring of CD4+Vbeta+ clonal T cells by fluorescence-activated cell sorting with double staining is a suitable method to assess clinical responses to different therapies.
覃样肉芽肿和 Sézary 综合征是皮肤 T 细胞淋巴瘤最常见的形式。为了评估对不同疗法的反应，尤其是在 Sézary 综合征中，监测血液中循环肿瘤细胞的百分比是有帮助的。使用 T 细胞受体（TCR）-Vβ特异性单克隆抗体为检测 Sézary 细胞提供了一种合适的工具。在本研究中，我们使用流式细胞术分析了 7 例接受各种治疗方式的患者的克隆性 CD4⁺Vβ⁺细胞水平，并通过用一组识别淋巴表面标志物的抗体进行双重染色来研究克隆细胞的免疫表型。此外，对克隆性 CD4⁺Vβ2⁺细胞进行了聚合酶链反应-变性梯度凝胶电泳分析，结果表明这些细胞携带 Vγ10/11、JγP1/2 TCR 重排。随访研究显示，6 例患者中 Vβ⁺克隆的发展与对不同疗法的临床反应密切相关。在 2 例患者中，CD4⁺Vβ⁺细胞减少，同时伴有部分缓解甚至完全缓解。在 4 例患者中，克隆性 CD4⁺Vβ⁺细胞群体稳定或增加很好地反映了疾病的稳定或进展过程。Vβ⁺细胞的双重染色显示出以下模式：CD3⁺、CD5⁺、CD7⁺、CD28⁺、CD80⁻、CD86⁺和人类白细胞抗原（HLA）I 类⁺。相比之下，HLA-DR 表达异质性。我们得出结论，通过双重染色的荧光激活细胞分选来鉴定和监测 CD4⁺Vβ⁺克隆性 T 细胞是评估对不同疗法临床反应的一种合适方法。
