Sales Maria M, Bezerra Camila N A, Hiraki Yumi, Melo Neusa B, Rebouças Nancy A
Serviço de Hematologia, Divisão de Laboratório Central, Departamento de Patologia/Hospital das Clínicas, Universidade de São Paulo, Av. Prof. Lineu Prestes 1524, 05508-900-São Paulo, SP, Brazil.
Eur J Haematol. 2005 Oct;75(4):280-7. doi: 10.1111/j.1600-0609.2005.00506.x.
The diagnosis of Adult T-cell leukemia/lymphoma ATLL subtypes in human T-lymphotropic virus type I (HTLV-I) carriers based in morphology and immunophenotype of lymphocytes can be challenger. We propose that polymerase chain reaction (PCR) amplification of the rearranged TCR gene in HTLV-I healthy carriers would be a convenient method for establishing the nature of the circulating T lymphocytes in asymptomatic HTLV-I carriers, presenting only mild and inconclusive signals of deviation from normality.
Using PCR, we analyzed the genetic recombination pattern of the T-cell beta-chain receptor gene (TCR-beta) in order to identify clonal expansion of peripheral blood T lymphocytes in 17 HTLV-I-positive healthy carriers and in nine normal HTLV-I-negative blood donors. To evaluate the performance of PCR in detection of clonality, we also analyzed 18 patients with post-thymic/mature T-cell malignancies presenting circulating abnormal lymphocytes.
Seven of the 17 HTLV-I positive individuals presented circulating abnormal lymphocytes; monoclonal or oligoclonal expansion of T-cells was detected in five of the 17 HTLV-I-positive individuals, all of them presenting abnormal lymphocytes. Clonal expansion was not detected in any of the negative controls or in any of the 12 remaining healthy carriers. All patients in the positive control group tested positive by PCR and Southern blots. Southern blots were negative for all 17 healthy carriers.
PCR amplification of segments of rearranged TCR-beta is reliable for allowing early detection of small populations of clonal T cells in blood samples from asymptomatic HTLV-I carriers, providing an additional alert in the follow-up of carriers with abnormal circulating lymphocytes.
基于淋巴细胞的形态学和免疫表型对I型人类嗜T淋巴细胞病毒(HTLV-I)携带者的成人T细胞白血病/淋巴瘤(ATLL)亚型进行诊断可能具有挑战性。我们提出,对HTLV-I健康携带者重排的TCR基因进行聚合酶链反应(PCR)扩增,将是一种便捷的方法,用于确定无症状HTLV-I携带者循环T淋巴细胞的性质,这些携带者仅表现出轻微且不确定的偏离正常的信号。
我们使用PCR分析了T细胞β链受体基因(TCR-β)的基因重组模式,以鉴定17名HTLV-I阳性健康携带者和9名正常HTLV-I阴性献血者外周血T淋巴细胞的克隆性扩增。为了评估PCR检测克隆性的性能,我们还分析了18例胸腺后/成熟T细胞恶性肿瘤患者,这些患者存在循环异常淋巴细胞。
17名HTLV-I阳性个体中有7名存在循环异常淋巴细胞;在17名HTLV-I阳性个体中的5名检测到T细胞单克隆或寡克隆扩增,他们均存在异常淋巴细胞。在任何阴性对照或其余12名健康携带者中均未检测到克隆性扩增。阳性对照组的所有患者通过PCR和Southern印迹检测均为阳性。17名健康携带者的Southern印迹均为阴性。
重排的TCR-β片段的PCR扩增对于早期检测无症状HTLV-I携带者血样中的少量克隆性T细胞是可靠的,为循环淋巴细胞异常的携带者的随访提供了额外的警示。