Immunophenotypic analysis of the TCR-Vβ repertoire in 98 persistent expansions of CD3+/TCR-αβ+ large granular lymphocytes -: Utility in assessing clonality and insights into the pathogenesis of the disease
Immunophenotypic analysis of the TCR-Vβ repertoire in 98 persistent expansions of CD3+/TCR-αβ+ large granular lymphocytes -: Utility in assessing clonality and insights into the pathogenesis of the disease
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DOI：
10.1016/s0002-9440(10)63032-5
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发表时间：
2001-11-01
影响因子：
6
通讯作者：
Orfao, A
中科院分区：
文献类型：
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作者：
Lima, M;Almeida, J;Orfao, A
目前，大颗粒淋巴细胞（LGL）白血病初步诊断的一个主要挑战是确定扩大群体的克隆性质。在本研究中，我们通过流式细胞术免疫表型分析了 98 个连续 CD4(+) 或 CD8(+bright)CD3(+)/TCR-alpha beta (+) LGL 持续扩增病例的 TCR-V beta 库，并将结果与 TCR-beta 基因重排的分子研究中获得的结果进行了比较。 58 例在分子研究中被认为是单克隆，而其余 40 例没有单克隆证据（11 例被认为是寡克隆，29 例被认为是多克隆）。在 96% 的病例中，TCR-V beta 库倾向于优先使用一个或多个 TCR-V beta 家族，总共诊断了 124 个 TCR-V beta 扩展：71 个病例中有 1 个 TCR-V beta 扩展，23 个病例有两个或多个 TCR-V beta 扩展。与非单克隆病例 (24 +/- 14%) (24 +/- 14%) (P = 0.001) 相比，在每个病例中观察到的最高 TCR-V β 扩增在单克隆病例中更高 (74 +/- 19%)，表现出 TCR-V β 限制模式的 LGL 比例也是如此 (分别为 86 +/- 16% 和 42 +/- 23%；P = 0.0001)；相比之下，后一组中显示不止一个 TCR-V β 扩增的病例比例较高：分别为 7% 和 48% (P = 0.001)。寡克隆病例中获得的结果介于多克隆和单克隆病例中获得的结果之间，并且对于 CD4(+) 和 CD8(+bright) T 细胞扩增观察到相似的结果。 CD8(+bright) T 细胞-LGL 增殖中表达的 TCR-V β 家族表现出一种分布模式，模拟单个 TCR-V β 家族在正常外周血 T 细胞中出现的频率。假设每当给定的 TCR-V β 家族扩增至少占总 CD4(+) 或 CD8(+bright) T 细胞的 40% 时，给定的 LGL 增殖是单克隆的。隔室中，我们能够以 93% 的敏感性和 80% 的特异性预测克隆性。通过将截止值提高到 60%，敏感性和特异性分别为 81% 和 100%。总之，我们的结果表明，TCR-V beta 库的流式细胞术免疫表型分析是评估 TCR-alpha beta (+) LGL 持续扩增中 T 细胞克隆性的强大筛选手段。
At present, a major challenge in the initial diagnosis of leukemia of large granular lymphocytes (LGLs) is to establish the clonal nature of the expanded population. in the present study we have analyzed by flow cytometry immunophenotyping the TCR-V beta repertoire of 98 consecutive cases of persistent expansions of CD4(+) or CD8(+bright)CD3(+)/TCR-alpha beta (+) LGLs and compared the results with those obtained in molecular studies of TCR-beta gene rearrangements. Fifty-eight cases were considered to be monoclonal in molecular studies whereas in the remaining 40 cases there was no evidence for monoclonality (11 cases were considered oligoclonal and 29 polyclonal). The TCR-V beta repertoire was biased to the preferential use of one or more TCR-V beta families in 96% of cases, a total of 124 TCR-V beta expansions being diagnosed: one TCR-V beta expansion in 71 cases and two or more TCR-V beta expansions in 23 cases. The highest TCR-V beta expansion observed in each case was higher among monoclonal (74 +/- 19%) as compared to nonmonoclonal cases (24 +/- 14%) (P = 0.001), as did the fraction of LGLs that exhibited a TCR-V beta -restricted pattern (86 +/- 16% and 42 +/- 23%, respectively; P = 0.0001); by contrast, the proportion of cases displaying more than one TCR-V beta expansion was higher in the latter group: 7% versus 48%, respectively (P = 0.001). Results obtained in oligoclonal cases were intermediate between those obtained in polyclonal and monoclonal cases and similar results were observed for CD4(+) as for CD8(+bright) T-cell expansions. TCR-V beta families expressed in CD8(+bright) T-cell-LGL proliferations showed a pattern of distribution that mimics the frequency at which the individual TCR-V beta families are represented in normal peripheral blood T cells. Assuming that a given proliferation of LGLs is monoclonal whenever there is an expansion of a given TCR-V beta family of at least 40% of the total CD4(+) or CD8(+bright) T-cell. compartment, we were able to predict clonality with a sensitivity of 93% and a specificity of 80%. By increasing the cut-off value to 60%, sensitivity and specificity were of 81% and 100%. In summary, our results suggest that flow cytometry immunophenotypic analysis of the TCR-V beta repertoire is a powerful screening toot for the assessment of T-cell clonality in persistent expansions of TCR-alpha beta (+) LGLs.