Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Cancer Biomark. 2018;22(4):733-745. doi: 10.3233/CBM-181295.
To find out if the T cell repertoire is efficiently and specifically provoked in patients with breast cancer, we have investigated the clonotypes of main T cell subsets (based on Vβ-Chain) in tumor draining lymph nodes. CD4+ helper, CD8+ cytotoxic and (CD4+CD127dimCD25+) regulatory T cells, were negatively selected, and isolated, from lymph node mononuclear cells of 14 untreated patients with breast cancer. Four non-malignant patients, who underwent surgical operation, were also recruited as the control group. Based on sequences and new nomenclature of the T cell Receptor β Variables (TRBVs) available in the international ImMunoGeneTics (IMGT) database, 28 TRBV specific forward primers and two TRB Constant region (TRBC) specific reverse primers were developed to amplify all functional alleles. Fluorescent-labeled PCR products were then run on an ABI PRISM 310 Genetic-Analyzer. The data was analyzed by GeneMapper software version 3.1. Clonotype analysis suggested that activated T cells are present in breast cancer. More TRBV usage were detected among CD4+ helper and regulatory subsets, with Gaussian-like pattern in the majority of functional TRBV families; whereas CD8+ cytotoxic T cells showed oligoclonality in almost all TRBV families with one or two dominant peaks in each family. Similar pattern in some of these TRBVs were also observed among controls. Having no expression or polyclonality in the controls, the oligoclonal pattern observed in the TRBV18, however, appears to be specific to breast cancer patients. This phenomenon may reflect the existence of new antigenic stimulation(s) in BC patients, preferentially activating those clones of T cells that express TRBV18.
为了探究乳腺癌患者的 T 细胞受体库是否能得到有效且特异性的激活，我们对肿瘤引流淋巴结中的主要 T 细胞亚群（基于 Vβ-链）的克隆型进行了研究。我们从 14 例未经治疗的乳腺癌患者的淋巴结单核细胞中，使用阴性选择和分离的方法，获得了 CD4+辅助性 T 细胞、CD8+细胞毒性 T 细胞和（CD4+CD127dimCD25+）调节性 T 细胞。我们还招募了 4 例接受手术的非恶性患者作为对照组。根据国际免疫遗传学（IMGT）数据库中 T 细胞受体 β 变量（TRBVs）的序列和新命名法，我们开发了 28 个 TRBV 特异性正向引物和两个 TRB 恒定区（TRBC）特异性反向引物，以扩增所有功能性等位基因。然后将荧光标记的 PCR 产物在 ABI PRISM 310 遗传分析仪上运行。使用 GeneMapper 软件版本 3.1 对数据进行分析。克隆型分析表明，乳腺癌患者中存在活化的 T 细胞。在 CD4+辅助性和调节性亚群中检测到更多的 TRBV 利用，大多数功能性 TRBV 家族呈现高斯样模式；而 CD8+细胞毒性 T 细胞在几乎所有的 TRBV 家族中都表现出寡克隆性，每个家族中只有一个或两个主要峰。在对照组中也观察到了一些这些 TRBV 的类似模式。在对照组中没有表达或多克隆性，然而，在 TRBV18 中观察到的寡克隆模式似乎是乳腺癌患者所特有的。这种现象可能反映了 BC 患者存在新的抗原刺激，优先激活表达 TRBV18 的 T 细胞克隆。
