Wu C J, Chillemi A, Alyea E P, Orsini E, Neuberg D, Soiffer R J, Ritz J
Center for Hematologic Oncology and Department of Biostatistics, Dana-Farber Cancer Institute, Boston, MA 02115, USA.
Blood. 2000 Jan 1;95(1):352-9.
CDR3 spectratyping was used to analyze the complexity of the T-cell repertoire and to define the mechanisms and kinetics of the reconstitution of T-cell immunity after allogeneic bone marrow transplantation (BMT). This method, which is based on polymerase chain reaction amplification of all CDR3 regions using the T-cell receptor (TCR) Vbeta genes, was used to examine serial samples of peripheral blood lymphocytes from 11 adult patients with chronic myelogenous leukemia (CML) who underwent T-cell-depleted allogeneic BMT. In contrast to 10 normal donors who display highly diverse and polyclonal spectratypes, patient samples before and early after BMT revealed markedly skewed repertoires, consisting of absent, monoclonal, or oligoclonal profiles for the majority of Vbeta subfamilies. To quantify changes in TCR repertoire over time, we established an 8-point scoring system for each Vbeta subfamily. The mean complexity score for patient samples before transplant (130.8) was significantly lower than that for normal donors (183; P = 0. 0007). TCR repertoire complexity was abnormal in all patients at 3 months after BMT (mean score = 87). Normalization of repertoire began in 4 patients at 6 months after BMT, but the majority of patients continued to display abnormal repertoires for up to 3 years after BMT. To determine whether the reconstituted T-cell repertoire was derived from the donor or recipient, unique microsatellite loci were examined to establish chimeric status. At 3 months after BMT, 7 patients demonstrated mixed chimerism; 4 had complete donor hematopoiesis (CDH). CDH strongly correlated with likelihood of restoration of T-cell repertoire complexity (P = 0.003). In contrast, patients who demonstrated persistence of recipient hematopoiesis failed to reconstitute a diverse TCR repertoire. These findings suggest that the reconstitution of a normal T-cell repertoire from T-cell progenitors in adults is influenced by interactions between recipient and donor hematopoietic cells. (Blood. 2000;95: 352-359)
互补决定区3（CDR3）谱型分析用于分析T细胞库的复杂性，并确定异基因骨髓移植（BMT）后T细胞免疫重建的机制和动力学。该方法基于使用T细胞受体（TCR）Vβ基因对所有CDR3区域进行聚合酶链反应扩增，用于检测11例接受T细胞去除的异基因BMT的成年慢性粒细胞白血病（CML）患者的外周血淋巴细胞系列样本。与10名显示高度多样化和多克隆谱型的正常供体相比，BMT前及BMT后早期的患者样本显示出明显偏态的库，大多数Vβ亚家族呈现缺失、单克隆或寡克隆图谱。为了量化TCR库随时间的变化，我们为每个Vβ亚家族建立了一个8分评分系统。移植前患者样本的平均复杂性评分（130.8）显著低于正常供体（183；P = 0.0007）。BMT后3个月时所有患者的TCR库复杂性均异常（平均评分 = 87）。4例患者在BMT后6个月时库开始正常化，但大多数患者在BMT后长达3年仍显示异常库。为了确定重建的T细胞库是来源于供体还是受体，检测了独特的微卫星位点以确定嵌合状态。BMT后3个月时，7例患者表现为混合嵌合；4例有完全供体造血（CDH）。CDH与T细胞库复杂性恢复的可能性密切相关（P = 0.003）。相反，显示受体造血持续存在的患者未能重建多样化的TCR库。这些发现表明，成体中T细胞祖细胞正常T细胞库的重建受受体和供体造血细胞之间相互作用的影响。（《血液》。2000年；95：352 - 359）
