Oligoclonal and polyclonal CD4 and CD8 lymphocytes in aplastic anemia and paroxysmal nocturnal hemoglobinuria measured by Vβ CDR3 spectratyping and flow cytometry
Oligoclonal and polyclonal CD4 and CD8 lymphocytes in aplastic anemia and paroxysmal nocturnal hemoglobinuria measured by Vβ CDR3 spectratyping and flow cytometry
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DOI：
10.1182/blood-2002-01-0236
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发表时间：
2002-07-01
期刊：
影响因子：
20.3
通讯作者：
Maciejewski, JP
中科院分区：
文献类型：
--
作者：
Risitano, AM;Kook, H;Maciejewski, JP
我们假设在再生障碍性贫血（AA）中，抗原特异性T细胞的存在通过它们对特定可变β链（Vbeta）亚家族的扩增的贡献以及通过克隆CDR 3偏斜来反映。为了确定疾病特异性“签名”T细胞克隆在AA中的作用，我们通过流式细胞术研究了Vbeta的优先使用，并分析了Vbeta-CDR 3区域的寡克隆性的存在。我们首先确定了每个V β家族对总CD 4（+）和CD 8（+）淋巴细胞池的贡献;在AA和阵发性睡眠性血红蛋白尿症中，观察到不同VP家族的随机过度表达。平均而言，我们发现扩展在3（22检查）Vbeta家庭每例患者。当检测单个Vbeta家族对效应子库的贡献时，发现了更显著的Vbeta偏置（skewing. Vbeta-CDR 3大小分布分析了分离的CD 4（+）和CD 8（+）群体中扩增的Vbeta家族;代表性不足的Vbeta家族显示出更明显的CDR 3偏置。扩增的CD 4（+）Vbeta亚家族大多显示多克隆CDR 3大小分布，在扩增的Vbeta家族中仅38%的偏斜。相反，在过度表达的CD 8（+）Vbeta类型中，观察到明显的CDR 3偏斜（82%），与特异性CD 8（+）T细胞克隆的非随机扩增一致。没有观察到与HLA型相关的特定Vbeta家族的优先扩展。在免疫抑制治疗后检查的患者中，异常的V β分布模式被保留，但单个V β的扩展程度较低。由于Vbeta偏斜可能与相对Vbeta大小相关，寡克隆性与数值Vbeta扩增的组合可用于识别疾病特异性T细胞受体。
We have hypothesized that in aplastic anemia (AA) the presence of antigen-specific T cells is reflected by their contribution to the expansion of a particular variable beta chain (Vbeta) subfamily and also by clonal CDR3 skewing. To determine the role of disease-specific "signature" T-cell clones in AA, we studied preferential Vbeta usage by flow cytometry and analyzed Vbeta-CDR3 regions for the presence of oligoclonality. We first established the contribution of each Vbeta family to the total CD4(+) and CD8(+) lymphocyte pool; in AA and paroxysmal nocturnal hemoglobinuria, a seemingly random overrepresentation of different VP families was observed. On average, we found expansion in 3 (of 22 examined) Vbeta families per patient. When the contribution of individual Vbeta families to the effector pool was examined, more striking Vbeta skewing was found. Vbeta-CDR3 size distribution was analyzed for the expanded Vbeta families in isolated CD4(+) and CD8(+) populations; underrepresented Vbeta families displayed more pronounced CDR3 skewing. Expanded CD4(+)Vbeta subfamilies showed mostly a polyclonal CDR3 size distribution with only 38% of skewing in expanded Vbeta families. In contrast, within overrepresented CD8(+)Vbeta types, marked CDR3 skewing (82%) was seen, consistent with nonrandom expansion of specific CD8(+) T-cell clones. No preferential expansion of particular Vbeta families was observed, in relation to HLA-type. In patients examined after immunosuppressive therapy, an abnormal Vbeta-distribution pattern was retained, but the degree of expansion of individual Vbeta was lower. As Vbeta skewing may correlate with relative Vbeta size, oligoclonality in combination with numerical Vbeta expansion can be applied to recognition of disease-specific T-cell receptors.