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Establishment and characterization of a novel Hodgkin lymphoma cell line, AM‐HLH, carrying the Epstein‐Barr virus genome integrated into the host chromosome

机译:新型霍奇金淋巴瘤细胞系,AM-HLH的建立和表征,将Epstein-Barr病毒基因组载入宿主染色体中

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Abstract We describe the establishment and characterization of a cell line, AM‐HLH, obtained from a patient with Epstein‐Barr virus–positive (EBV + ) nodular sclerosis–type Hodgkin lymphoma (HL). The cells were positive for CD2 and CD30 and negative for CD15. The immunoglobulin heavy‐ and κ light–chain genes were rearranged. The karyotype was of the triploid range. Southern blotting using the EBV terminal repeat probe detected 3 hybridizing bands that were identical to those of the parental HL material. The cells expressed EBV‐encoded RNAs as well as latent genes ( EBNA1 , EBNA2 , LMP1 , and LMP2A ) and lytic genes ( BZLF1 and BALF2 ). Fluorescence in situ hybridization (FISH) with the cosmid pJB8 clone containing a fragment of EBV DNA as a probe revealed multiple hybridization signals at a marker chromosome. Additional FISH using whole chromosome painting and centromere probes in combination with multicolor FISH determined that multiple EBV copies were clustered within the chromosome 20 materials of the marker chromosome. Culture supernatants of AM‐HLH contained IL‐10 as measured by the bead‐based immunoassay. It is possible that an integrated EBV genome and cellular genes on chromosome 20 were coamplified, leading to the enhanced expression of genes involved in cell growth control. The AM‐HLH cell line will be useful to clarify the role of cytokines in the development of EBV + HL.
机译:摘要我们描述了从一例EB病毒阳性(EBV+)结节性硬化型霍奇金淋巴瘤(HL)患者中获得的细胞系AM-HLH的建立和特征。CD2和CD30阳性,CD15阴性。免疫球蛋白重链和κ轻链基因被重新排列。核型为三倍体。使用EBV末端重复探针进行Southern杂交,检测到3条与亲本HL材料相同的杂交条带。这些细胞表达EBV编码的RNA以及潜伏基因(EBNA1、EBNA2、LMP1和LMP2A)和裂解基因(BZLF1和BALF2)。以含有EBV DNA片段的cosmid pJB8克隆为探针的荧光原位杂交(FISH)在标记染色体上显示了多种杂交信号。使用全染色体绘制和着丝粒探针结合多色FISH的其他FISH确定,多个EBV拷贝聚集在标记染色体的20号染色体材料中。AM-HLH的培养上清液中含有IL-10,通过基于珠的免疫分析测定。整合的EBV基因组和20号染色体上的细胞基因可能被共扩增,导致参与细胞生长控制的基因表达增强。AM-HLH细胞系将有助于阐明细胞因子在EBV+HL发展中的作用。

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