Department of Hematopathology, University of Texas MD Anderson Cancer Center, Houston, TX 77030-4009, USA.
Mod Pathol. 2010 Feb;23(2):284-95. doi: 10.1038/modpathol.2009.175. Epub 2009 Nov 20.
Peripheral blood involvement has been recognized as an adverse prognostic factor in patients with mycosis fungoides and Sézary syndrome. However, accurate identification and enumeration of the neoplastic cells in these diseases can be challenging. We assessed the clinical utility of flow cytometric immunophenotypic analysis of T-cell receptor Vbeta expression in 82 mycosis fungoides and 6 Sézary syndrome patients, with an atypical T-cell immunophenotype, or abnormal CD4:CD8 ratio, identified from peripheral blood specimens of 723 patients submitted for routine mycosis fungoides/Sézary syndrome blood staging. To improve detection sensitivity, Vbeta expression was analyzed on gated CD3+CD4+ T cells or T cells with an aberrant immunophenotype, if present. The flow cytometric results were compared with traditional morphologic assessment (n=88) and molecular methods to assess the T-cell receptor gamma or beta genes (n=41 tested in parallel). Flow cytometric immunophenotyping yielded a clonal Vbeta pattern in 60/82 mycosis fungoides and 6/6 Sézary syndrome patients. By contrast, flow cytometric Vbeta was negative in all 10 healthy donors and 18 control patients, showing a specificity of 100% and concordance with molecular testing of 86%. Using flow cytometric Vbeta results instead of morphologic assessment, 12 patients were upstaged from B1 to B2, and 20 patients from B0 to B1 (P<0.0001). The 12 upstaged B2 patients had no morphologic evidence of involvement, but had an aggressive clinical course similar to those staged by traditional morphologic assessment (median survival 27 vs 41 months, log-rank P=0.701). In 30/44 patients with a tumor-associated Vbeta expression, a single Vbeta tube was used to monitor treatment response. In conclusion, flow cytometric Vbeta analysis is rapid and convenient, can assess T-cell clonality and tumor quantity simultaneously, and is useful both in initial blood staging and monitoring tumor burden during therapy in patients with mycosis fungoides or Sézary syndrome.
外周血受累已被认为是蕈样肉芽肿和塞扎里综合征患者的不良预后因素。然而，准确识别和计数这些疾病中的肿瘤细胞可能具有挑战性。我们评估了流式细胞术免疫表型分析 T 细胞受体 Vβ 表达在 82 例蕈样肉芽肿和 6 例塞扎里综合征患者中的临床应用，这些患者的外周血标本具有非典型 T 细胞免疫表型或异常 CD4:CD8 比值，这些患者是从 723 例常规蕈样肉芽肿/塞扎里综合征血液分期患者的标本中识别出来的。为了提高检测灵敏度，如果存在异常免疫表型的 T 细胞，则对 gated CD3+CD4+T 细胞或 T 细胞进行 Vβ 表达分析。将流式细胞术结果与传统形态学评估（n=88）和评估 T 细胞受体 γ 或β 基因的分子方法（n=41 例平行检测）进行比较。流式细胞术免疫表型在 60/82 例蕈样肉芽肿和 6/6 例塞扎里综合征患者中产生了克隆性 Vβ 模式。相比之下，所有 10 名健康供体和 18 名对照患者的流式细胞术 Vβ 均为阴性，特异性为 100%，与 86%的分子检测结果一致。使用流式细胞术 Vβ 结果替代形态学评估，12 例患者从 B1 期升级为 B2 期，20 例患者从 B0 期升级为 B1 期（P<0.0001）。12 例升级为 B2 期的患者没有形态学受累的证据，但具有与传统形态学评估相似的侵袭性临床病程（中位生存时间 27 个月与 41 个月，对数秩 P=0.701）。在 30/44 例具有肿瘤相关 Vβ 表达的患者中，使用单个 Vβ 管监测治疗反应。总之，流式细胞术 Vβ 分析快速方便，可同时评估 T 细胞克隆性和肿瘤数量，在蕈样肉芽肿或塞扎里综合征患者的初始血液分期和监测治疗期间肿瘤负荷方面均有用。
