摘要背景 近年来眼眶淋巴瘤的发病率逐渐升高,在病理特点、治疗方法以及发病机制方面的研究不断深入.由于眼眶淋巴瘤发病率低,体外培养困难,眼眶淋巴瘤裸鼠模型研究的报道较少. 目的 应用细胞系pfeiffer建立弥漫大B细胞淋巴瘤(DLBCL)眼眶裸鼠模型,比较小鼠淋巴瘤模型与人眼眶DLBCL病理标本的病理学特点及生物学行为,探讨全身DLBCL细胞系pfeiffer用于眼眶淋巴瘤研究的可能性.方法 选用SPF级BALB/c裸鼠10只和nod-SCID小鼠5只,先用137 Cs对BALB/c裸鼠(吸收剂量为3.5 Gy)和nod-SCID小鼠(吸收剂量为2.6Gy)进行照射,照射后6h内分别进行小鼠眼眶内(BALB/c小鼠4只眼)及皮下注射(BALB/c小鼠和nod-SCID小鼠各4只眼)pfeiffer细胞,接种细胞密度约为1.5×108/ml,即眼眶接种组和皮下接种组.注射后每日观察肿瘤的生长状态,并绘制肿瘤生长曲线.于注射后54 d无菌条件下完整取出小鼠眼眶肿瘤和皮下肿瘤及附近淋巴结,制备4μm厚组织切片.采用苏木精-伊红染色评估肿瘤的组织病理学特点,采用免疫组织化学法检测模型肿瘤切片中CD20、CD79α、CD45RO蛋白的表达,并根据CD10、BCL-6和mum-1的表达进行分型,依据Ki-67及survivin的表达判断肿瘤的预后,并将模型小鼠的检测结果与人眼眶DLBCL病理标本的特点进行比较.结果 眼眶接种组和皮下接种组成瘤率均为100％,nod-SCID小鼠的肿瘤生长速度快于BALB/c裸鼠.组织病理学检查可见眼眶接种组小鼠邻近淋巴结无肿瘤细胞浸润,皮下接种组小鼠腋窝淋巴结少量肿瘤细胞浸润.苏木精-伊红染色显示,小鼠淋巴瘤组织的组织病理学特点与人眼眶DLBCL标本一致.BALB/c小鼠CD20表达强度＜50％和≥50％的淋巴瘤模型中表达例数分别为3和5,CD79α分别为2和6,CD45RO分别为8和0;nod-SCID小鼠CD20表达强度＜50％和≥50％的淋巴瘤模型中表达例数分别为1和3,CD79α分别0和4,CD45RO分别为4和0;与人眼眶DLBCL标本的1和2,1和2,2和1比较,差异均无统计学意义(均P=1.00);BALB/c和nod-SCID小鼠淋巴瘤模型与人眼眶DLBCL中Ki-67和survivin表达的例数差异均无统计学意义(均P=1.00).BALB/c小鼠、nod-SCID小鼠淋巴瘤模型和人眼眶DLBCL中依据CD10、BCL-6和mum-1的表达均分为非生发中心来源型. 结论 采用pfeiffer细胞系接种法可成功建立眼眶和皮下DLBCL的小鼠模型,这些模型肿瘤在生物学行为、病理学特点和免疫组织化学染色方面均一致.Nod-SCID小鼠较BALB/c小鼠接种后肿瘤生长更快.

abstractsBackground Recently, the morbidity of orbital lymphoma increased gradually, and we have made deeper research in pathology,therapy and pathogenesis of the disease.There were few reports of mice model of orbital lymphoma up to now for its lower morbidity and culture difficulty.Objective This study was to establish a mouse model of orbital diffuse large B cell lymphoma (DLBCL) by injection of systemic DLBCL cell line pfeiffer.Methods Ten SPF BALB/c mice and 5 nod-SCID mice were radiated firstly using 137Cs,with the absorption dose 3.5 Gy in the BALB/c mice and 2.6 Gy in the nod-SCID mice,and then pfeiffer cells were intraobitally injected in 4eyes of BALB/c mice (orbital injection group) and suncutaneously injectied in 4 eyes of BALB/c mice and 4 eyes of nod-SCID mice (subcutaneous injection group) at the concentration of 1.5×l08/ml.The developing status of tumors were examined once per day and the growth curve was drawn.The tumors and nearby lymph nodes were obtained 54 days after injection for the preparation of 4 μm thickness of serial sections.Hemotoxylin-eosin staining was used to examine the histopathology of the tumors, and immunochemistry was employed to detect the expressions of CD20,CD79α and CD45RO proteins.The tumors were typed based on the expressions of CD10, BCL-6 and mum-1 in the specimens,and the expressions of Ki-67 and survivin were assyed to assess the prognosis of the tumor.All the results were compared with 3 diagnosed human orbital DLBCL sections.The use and care of the mice complied with Chinese Administration Rule of Laboratory Animal.Results The tumor formation rates were 100％ in both the orbital injection group and subcutaneous injection group, and the tumors grew much faster in nod-SCID mice than BALB/c mice.Infiltration of tumor cells in lymph nodes were found in the subcutaneous injection group rather than the orbital injection group.The pathological features were accordant among the orbital injection group, subcutaneous injection group and human orbital DLBCL sections.The number of ＜50％ and ≥50％ CD20+ specimens was 3 and 5,CD79αwas 2 and 6,CD45RO was 8 and 0 in the BALB/c mice;while that in the nod-SCID mice was 1 and 3 in CD20,0 and 4 in CD79α,4 and 0 in CD45RO;the number of human orbital DLBCL specimens was 1 and 2 in CD20,1 and 2 in CD79α,2 and 1 in CD45RO,without significant differences among them (all at P=1.00).No significant differences were seen in Ki-67+ number and survivin+ number among the BALB/c mice, nod-SCID mice and human orbital DLBCL specimens (all at P=1.00).The detection of CD10,BCL-6 and mum-1 expressions indicated that the tumors of BALB/c mice,nod-SCID mice and human orbital DLBCL specimens all were the non-germinal center B cell-like types.Conclusions The orbital and subcutaneous DLBCL mouse models are successfully established by injection of pfeiffer cell line.There are the same findings and features in biological behavior, pathology and immunohistochemistry in orbital,subcutaneous models with human orbitl DLBCL.Nod-SCID mice appear to be more suitable for the growth of lymphoma cells.