Chen Zhong, Tan Wan-long, Huang Xin, Liang Zhong-kun, Xu Cui-xiang, Gao Ji-min
Department of Urologic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2010 May;30(5):936-40.
To investigate a novel immunotherapy through immobilization of streptavidin-tagged hTNF-alpha on the biotinylated mucosal surface of the bladder wall for bladder cancer treatment in mice.
A total of 120 female C57BL/6j mice were randomized into 5 equal groups, namely blank control, PBS, soluble hTNF-alpha, SA-GFP, and SA-hTNF-alpha treatment groups. Twenty-four hours after establishment of a mouse model of orthotopic superficial bladder cancer, SA-hTNF-alpha fusion protein was immobilized on the biotinylated mucosal surface of the bladder wall, which was repeated every 4 days for a total of 6 sessions. Immunohistochemistry was performed to detect the retention time of SA-hTNF-alpha fusion protein in the biotinylated mouse bladder mucosa and the distribution of CD4(+) and CD8(+) lymphocytes in the mucosa and tumor tissues, with the tumor growth and mouse survival also observed. The cytotoxiciy of the tumor-specific lymphocytes was evaluated. The mice responding well to the treatment were re-challenged by MB49 and monitored for survival.
SA-hTNF-alpha could be efficiently and stably immobilized on the bladder mucosal surface for as long as 7 days. On day 60 after MB49 implantation, 18 out of 22 SA- hTNF-alpha-treated mice survived, with 9 appearing tumor-free, but all the mice in PBS control group died. Five out of 9 tumor-free mice in SA-hTNF-alpha group showed resistance to a re-challenge with intravesical MB49. The numbers of CD4(+) and CD8(+) lymphocytes were significantly greater in SA-hTNF-alpha group than in the other groups (P<0.05). The cytotoxicity of the tumor-specific lymphocytes was significantly stronger in SA-hTNF-alpha group than in the other groups (P<0.05).
SA-hTNF-alpha immobilized on the biotinylated mucosal surface of the bladder wall can significantly inhibit the tumor growth and promote the survival of the mice bearing orthotopic superficial bladder cancer.
研究一种通过将链霉亲和素标记的人肿瘤坏死因子-α（hTNF-α）固定在膀胱壁生物素化黏膜表面来治疗小鼠膀胱癌的新型免疫疗法。
将120只雌性C57BL/6j小鼠随机分为5组，即空白对照组、PBS组、可溶性hTNF-α组、SA-GFP组和SA-hTNF-α治疗组。建立原位浅表性膀胱癌小鼠模型24小时后，将SA-hTNF-α融合蛋白固定在膀胱壁生物素化黏膜表面，每4天重复一次，共进行6次。进行免疫组织化学检测SA-hTNF-α融合蛋白在生物素化小鼠膀胱黏膜中的保留时间以及黏膜和肿瘤组织中CD4(+)和CD8(+)淋巴细胞的分布，同时观察肿瘤生长情况和小鼠存活情况。评估肿瘤特异性淋巴细胞的细胞毒性。对治疗反应良好的小鼠用MB49进行再次攻击并监测其存活情况。
SA-hTNF-α可有效且稳定地固定在膀胱黏膜表面长达7天。在植入MB49后第60天，22只接受SA-hTNF-α治疗的小鼠中有18只存活，其中9只无肿瘤，但PBS对照组的所有小鼠均死亡。SA-hTNF-α组9只无肿瘤的小鼠中有5只对膀胱内注射MB49的再次攻击表现出抗性。SA-hTNF-α组中CD4(+)和CD8(+)淋巴细胞的数量明显多于其他组（P<0.05）。SA-hTNF-α组中肿瘤特异性淋巴细胞的细胞毒性明显强于其他组（P<0.05）。
固定在膀胱壁生物素化黏膜表面的SA-hTNF-α可显著抑制原位浅表性膀胱癌荷瘤小鼠的肿瘤生长并提高其存活率。
