Gu Linhui, Hong Lianlian, Ling Zhiqiang, Feng Jianguo, Zheng Zhiguo, Du Lingbin, Mou Hanzhou, Sun Wenyong, Kong Xiangming, Ling Yutian, Jiang Zhiming, Zhu Chihong, Mao Weimin, Qian Lijuan
Clin Lab. 2015;61(7):731-9. doi: 10.7754/clin.lab.2014.140602.
CD20 positive NK/T-cell lymphoma is extremely rare and difficult for clinical treatment. Due to the lack of an established cell model for this disease, less is known about its biological characterization and potential therapeutic options.
A cell line of NK/T-cell lymphoma, which was enriched by magnetic sorting with proper cell surface markers (CD56) from peripheral blood mononuclear cells (PBMCs) drawn from a 21-year-old male patient with nasal angiocentric NK/T-cell lymphoma, was designated as ZQNK-29. Immunophenotypic analysis of ZQNK-29 was performed by flow cytometric and immunohistochemical analysis. Comparative genomic hybridization (CGH) analysis was used for cytogenetic analysis of ZQNK-29. Potential rearrangements of the immunoglobulin gene and Epstein-Barr virus (EBV) infection were examined by PCR and RT-PCR, respectively.
ZQNK-29 cells express the phenotypic T-cell marker (CD3), T cell activation markers (HLA-DR), markers for both NK and cytotoxic T lymphocytes (TIA-1), and B-lineage marker CD20; however, expression of CD56 was not detected in expanded ZQNK-29 cells although this NK cell surface marker was used as one of selective cell surface markers for the initial isolation of NK/T cells. RT-PCR analysis showed that the pattern of gene expressions for infected EBV was latency type III, with the expressions of LMP1, EBNA-1, and EBNA-2; no rearrangements were found in the heavy-chain of the immunoglobulin gene or in the y chain of the T cell receptors (TCRs) gene. CGH analysis demonstrated that ZQNK-29 possessed an abnormal karyotype, 46XY, 1p (dist)+, 4p (dist)+, 4q (mid)-, 5q (mid)-, 9q (dist)+, 16p (dist)+, 16q (dist)+, 17p+, 17q (dist)+, 19q (dist)+, 20p+, 20q+, 21q+, and 22q+. Of these, 1p (dist)+, which has been confirmed to be mitochondrial DNA amplification, is believed to be mainly caused by EBV infection.
ZQNK-29 is a well characterized premature human NK/T-cell lymphoma cell line with expression of the B-cell marker CD20 and will provide a useful pre-clinic model for characterization and potential therapeutic studies of the aggressive NK/T-cell lymphoma.
CD20阳性NK/T细胞淋巴瘤极为罕见，临床治疗困难。由于缺乏该疾病成熟的细胞模型，对其生物学特性和潜在治疗方案的了解较少。
从一名21岁男性鼻型血管中心性NK/T细胞淋巴瘤患者外周血单个核细胞（PBMC）中，通过磁珠分选结合合适的细胞表面标志物（CD56）富集得到一株NK/T细胞淋巴瘤细胞系，命名为ZQNK-29。通过流式细胞术和免疫组化分析对ZQNK-29进行免疫表型分析。采用比较基因组杂交（CGH）分析对ZQNK-29进行细胞遗传学分析。分别通过PCR和RT-PCR检测免疫球蛋白基因的潜在重排和EB病毒（EBV）感染情况。
ZQNK-29细胞表达表型T细胞标志物（CD3）、T细胞活化标志物（HLA-DR）、NK和细胞毒性T淋巴细胞标志物（TIA-1）以及B系标志物CD20；然而，尽管最初分选NK/T细胞时使用了CD56作为选择性细胞表面标志物之一，但在扩增的ZQNK-29细胞中未检测到CD56表达。RT-PCR分析显示，感染的EBV基因表达模式为Ⅲ型潜伏，伴有LMP1、EBNA-1和EBNA-2表达；免疫球蛋白基因重链和T细胞受体（TCR）基因的γ链均未发现重排。CGH分析表明，ZQNK-29具有异常核型，46XY，1p（远端）+，4p（远端）+，4q（中间）-，5q（中间）-，9q（远端）+，16p（远端）+，16q（远端）+，17p+，17q（远端）+，19q（远端）+，20p+，20q+，21q+，22q+。其中，已证实1p（远端）+为线粒体DNA扩增，认为主要由EBV感染引起。
ZQNK-29是一株特征明确的人NK/T细胞淋巴瘤早期细胞系，表达B细胞标志物CD20，将为侵袭性NK/T细胞淋巴瘤的特性研究和潜在治疗研究提供有用的临床前模型。
