^(125)Ⅰ标记rLTB-UreB及口服BALB/c小鼠示踪实验研究

In Vivo Distribution of 125I - labeled rLTB - UreB in Mice Administered with the Drug by Oral Route

目的 观察^(125)Ⅰ标记基因重组融合蛋白LTB-UreB灌喂BALB/c小鼠后的体内分布。方法 采用氯胺T氧化法标记rLTB-UreB,Sephadex-G25凝胶过滤纯化,纸层析鉴定放射化学纯度及比活度。将BALB/c小鼠随机分为PBS口服空白对照组、Na^(125) Ⅰ口服对照组和^(125)Ⅰ-rLTB-UreB实验组,于不同时间采集标本,γ放射计数器检测CPM(每分钟计数值),SPSS分析结果。结果 纯化后的标记蛋白纯度>90％。实验组PP结和肠系膜淋巴结CPM升高,与对照组比较差异有非常显著意义。结论 ^(125)Ⅰ标记融合蛋白口服后可在肠道粘膜下淋巴组织沉积,为融合蛋白作为幽门螺杆菌口服疫苗候选抗原提供重要实验依据。

Objective To study the in vivo distribution of 125I - labeled rLTB - UreB in mice administered with the drug by oral route. Methods rLTB - UreB was labeled with 1251 by chloramines - T method and purified by Sephadex G25 gel filtration, then detected for radiochemical purity and specific activity by paper chromatography. BALB/c mice were divided into 3 groups and treated with PBS, Na 125I and 125I - labeled rLTB - UreB by oral route respectively. Collect the specimens of blood and various organs of mice at different time after administration and detect for CPM by 7 - radioactive counter. Analyze the result by SPSS software. Results The purity of the 125I - labeled rLTB - UreB after purification reached above 90% . The CPM in Payer' s patches and mesenteric lymph nodes of the mice treated with 125I - labeled rLTB - UreB were significant higher than those of the 2 control groups. Conclusion 125I - labeled rLTB - UreB can deposit in mucosal lym-phoid tissue. The study provided an important basis for using the rLTB - UreB as a candidate of oral Helico-bacter pylori vaccine.