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EB病毒 (EBV ,Epstein Barrvirus)与人类多种肿瘤有关 ,尤其是与鼻咽癌和淋巴瘤的关系密切。为此研究了EB病毒在huPBL SCID嵌合体小鼠体内诱发人B细胞淋巴瘤的分子特性及肿瘤发生机制。从健康成人外周血分离出淋巴细胞 ,将之移植到SCID小鼠腹腔内 ,实验感染EBV ,观察肿瘤的形成 ;采用单向免疫扩散法连续监测小鼠血清中人IgG的含量。分别用PCR方法检测肿瘤组织中是否存在人Alu序列 ,原位杂交检测肿瘤组织中EB病毒小RNA分子EBER 1;免疫组织化学方法检测人白细胞分化抗原 (CD4 5、CD2 0、CD4 5RO、CD3) ,病毒基因(LMP1、EBNA2、BZLF1)的表达 ,以及细胞瘤基因蛋白 (p5 3、C myc、Bcl 2、Bax)在诱发肿瘤中的表达情况。结果发现 ,实验中 34只感染EBV的huPBL SCID小鼠有 2 4只诱发出肿瘤 ,根据病理形态学特征、Alu PCR和免疫标志均证实诱发瘤是人源性B淋巴细胞肿瘤。原位分子杂交显示肿瘤细胞核内存在EBER 1,少数瘤细胞表达EB病毒BZLF1蛋白阳性 ,部分瘤细胞表达LMP1和EBNA2蛋白阳性。连续监测 12只huPBL SCID小鼠血清中人IgG含量 ,发现IgG水平随诱瘤时间延长和肿瘤生长有逐渐增高趋势。免疫组化显示诱发的 2 4例淋巴瘤组织p5 3、C myc、Bcl 2和Bax蛋白表达阳性率分别为 83.33%、10 0 %、95 .83%、91.6 7%。结果
Epstein-Barr virus (Epstein-Barr virus) is associated with a variety of human cancers, especially nasopharyngeal carcinoma and lymphomas. To investigate the molecular characteristics and tumorigenic mechanism of Epstein-Barr virus (EBV) induced human B cell lymphoma in huPBL SCID chimeric mice. Lymphocytes were isolated from healthy adult peripheral blood and transplanted into the peritoneal cavity of SCID mice. EBV was infected experimentally to observe the formation of tumors. The serum IgG levels in mice were continuously monitored by one-way immunodiffusion. The human Alu sequences were detected by PCR and EBER 1 in the tumor tissues were detected by in situ hybridization. The differentiation of human leukocyte antigen (CD4 5, CD20, CD4 5RO and CD3) was detected by immunohistochemical method. , The expression of viral genes (LMP1, EBNA2, BZLF1), and the expression of oncogene proteins (p5 3, C myc, Bcl 2, Bax) in induced tumors. The results showed that 24 of huPBL SCID mice infected with EBV in the experiment had 24 tumor-inducing tumors. According to the pathological features, both Alu PCR and immunological markers confirmed that the induced tumor was a human B lymphocyte tumor. In situ hybridization revealed the presence of EBER 1 in the nucleus of tumor cells. A few tumor cells expressed positive for Epstein-Barr virus BZLF1 protein and some tumor cells expressed LMP1 and EBNA2 protein positive. The serum levels of human IgG in 12 huPBL SCID mice were continuously monitored. It was found that IgG level increased gradually with the time of tumorigenesis and tumor growth. Immunohistochemistry showed that the positive rates of p53, C-myc, Bcl-2 and Bax expression in 24 cases of lymphoma were 83.33%, 100%, 95.83% and 91.67% respectively. result