Lymphoblastoid cell line with B1 cell characteristics established from a chronic lymphocytic leukemia clone by in vitro EBV infection
Lymphoblastoid cell line with B1 cell characteristics established from a chronic lymphocytic leukemia clone by in vitro EBV infection
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DOI：
10.4161/onci.1.1.18400
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发表时间：
2012-01-01
期刊：
影响因子：
7.2
通讯作者：
Klein, Eva
中科院分区：
文献类型：
--
作者：
Rosen, Anders;Bergh, Ann-Charlotte;Klein, Eva
慢性淋巴细胞白血病（CLL）细胞表达EB病毒（EBV）受体，并可在体外感染。在大多数实验中，感染的细胞不表达EBV编码基因的生长促进组，因此它们不产生LCL。然而，对于例外的克隆，获得了品系。我们描述了一个新的线，HG 3，建立了体外EBV感染从IGHV 1 -2未突变的CLL患者克隆。所有细胞都表达EBNA-2和LMP-1，这是EBV编码的转化关键基因。该细胞系和患者的离体克隆的核型、FISH细胞遗传学和SNP阵列谱显示双等位基因13 q14缺失，伴有DLEU 7、miR 15 a/miR 16 -1的基因组缺失，这两种微小RNA在50%的CLL病例中缺失。CLL细胞的其他特征是：表达CD 5/CD 20/CD 27/CD 43和释放与凋亡细胞上的oxLDL样表位反应的IgM天然抗体（参见图11）。定型子集-1）。与从正常B细胞建立的两个LCL的比较显示32个基因以更高水平（> 2倍）表达。其中包括LHX 2和LILRA。这些基因可能在疾病的发展中发挥作用。LHX 2表达在自我更新的多能造血干细胞中显示，LILRA 4编码骨髓基质细胞抗原-2的受体，其有助于B细胞发育。24个基因表达水平较低，其中PARD 3是不对称细胞分裂所必需的。这些基因可能有助于通过调节造血区室中的细胞表型来建立CLL克隆的前体。CD 5/CD 20/CD 27/CD 43的表达和天然抗体的自发产生可以将CLL细胞鉴定为自我更新的B1淋巴细胞。
Chronic lymphocytic leukemia (CLL) cells express the receptor for Epstein-Barr virus (EBV) and can be infected in vitro. Infected cells do not express the growth-promoting set of EBV-encoded genes and therefore they do not yield LCLs, in most experiments. With exceptional clones, lines were obtained however. We describe a new line, HG3, established by in vitro EBV-infection from an IGHV1-2 unmutated CLL patient clone. All cells expressed EBNA-2 and LMP-1, the EBV-encoded genes pivotal for transformation. The karyotype, FISH cytogenetics and SNP-array profile of the line and the patient's ex vivo clone showed biallelic 13q14 deletions with genomic loss of DLEU7, miR15a/miR16-1, the two micro-RNAs that are deleted in 50% of CLL cases. Further features of CLL cells were: expression of CD5/CD20/CD27/CD43 and release of IgM natural antibodies reacting with oxLDL-like epitopes on apoptotic cells (cf. stereotyped subset-1). Comparison with two LCLs established from normal B cells showed 32 genes expressed at higher levels (> 2-fold). Among these were LHX2 and LILRA. These genes may play a role in the development of the disease. LHX2 expression was shown in self-renewing multipotent hematopoietic stem cells, and LILRA4 codes for a receptor for bone marrow stromal cell antigen-2 that contributes to B cell development. Twenty-four genes were expressed at lower levels, among these PARD3 that is essential for asymmetric cell division. These genes may contribute to establish precursors of CLL clones by regulation of cellular phenotype in the hematopoietic compartment. Expression of CD5/CD20/CD27/CD43 and spontaneous production of natural antibodies may identify the CLL cell as a self-renewing B1 lymphocyte.