减毒鼠伤寒杆菌为载体的幽门螺杆菌尿素酶口服疫苗的制备  

CONSTRUCTION OF AN ORAL VACCINE OF HELICOBACTER PYLORI UREASE SUBUNIT B WITH ATTENUATED SALMONELLA TYPHIMURIUM AS VECTOR

作　　者：刘晓峰[1] 胡家露[2] 张霞[1] 权启镇[1] 樊代明[2] 

机构地区：[1]济南军区总医院,济南250031 [2]第四军医大学西京医院全军消化病研究所

出　　处：《解放军医学杂志》2003年第7期633-635,共3页Medical Journal of Chinese People's Liberation Army

摘　　要：构建能表达幽门螺杆菌 (H .pylori)尿素酶B亚单位 (UreB)的重组减毒鼠伤寒杆菌SL3 2 61/ pTC0 1 UreB ,Westernblot检测UreB的表达。将SL3 2 61/pTC0 1 UreB口服免疫BALB/c小鼠 ,12周后检测肠液和血清中的特异性抗体反应及脾细胞培养液上清中的IFN γ和IL 10含量。结果显示 ,减毒鼠伤寒杆菌SL3 2 61/pTC0 1 UreB能表达约 61kD的蛋白 ,与H .pyloriUreB亚单位相符 ,具有抗原性。口服免疫小鼠后 ,疫苗组小鼠的肠液和血清中可分别检测到针对UreB的特异性IgA和IgG抗体 ,小鼠脾细胞培养上清中的IFN γ和IL 10的含量亦明显升高。病理学检查显示疫苗组小鼠较对照组小鼠胃粘膜炎症指数无差异。提示表达H .pyloriUreB的减毒鼠伤寒杆菌SL3 2 61/pTC0 1 UreB有望用作抗H .pylori感染的口服疫苗 ,其防治H .In order to construct a recombinant attenuated Salmonella typhimurium expressing Helicobacter pylori (H. pylori) urease subunit B(UreB), UreB gene fragment amplified by PCR was cloned into the prokaryotic expression vector pTC01 after sequencing, then transformed into attenuated Salmonella typhimurium SL3261 to acquire SL3261/pTC01-UreB. The expression of H. pylori UreB in SL3261 was detected by Western blot. Tewelve weeks after oral immunization of mice with this vaccine, anti-UreB IgA antibodies in mouse intestinal fluid and IgG antibodies in serum were determined by ELISA. IFN-γ and IL-10 contents in the supernatant of spleen cell culture were also assessed by ELISA. The results showed that 61kD protein was expressed in SL3261/pTC01-UreB that could be recognized by anti-H. pylori UreB antiserum by Western-blot. The multiple oral immunizations with SL3261/pTC01-UreB could induce significantly H. pylori-specific mucosal IgA response as well as serum IgG responses. Moreover, there was significant increase of IFN-γ and IL-10 contents in the supernatant of spleen cell culture in SL3261/pTC01-UreB group. These results suggested that the attenuated Salmonella typhimurium expressing H. pylori UreB may be used as oral vaccine against H. pylori infection. Its effect against H. pylori infection also needs to be further evaluated in animal models.

关 键 词：螺杆菌 幽门 尿素酶 沙门氏菌 鼠伤寒 疫苗 

分 类 号：R573.03[医药卫生—消化系统]