弓形虫ESA鼻内免疫小鼠诱导的黏膜及系统免疫应答  

Intranasal immunization with Toxoplasma gondii excreted-secreted antigene induced mucosal and systemic immune response in mice

作　　者：管志玉[1] 孟晓丽[1] 殷国荣[1] 刘红丽[1] 王海龙[1] 

机构地区：[1]山西医科大学医学寄生虫学研究所寄生虫学教研室,太原030001

出　　处：《山西医科大学学报》2007年第10期868-871,共4页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(30640057);山西省自然科学基金资助项目(20041105);山西省高校科技研究开发基金重点资助项目(20041238);山西省青年科技基金资助项目(20051045)

摘　　要：目的观察弓形虫排泄-分泌抗原(excreted-secreted antigen,ESA)鼻内免疫小鼠诱导黏膜和系统免疫应答的效果。方法将64只BALB/c小鼠随机分为8个组,每组8只。实验组小鼠分别用腹腔感染弓形虫小鼠第0,6,12,24,36,48和60小时无菌收集的腹腔液中获取的ESA 20μg/只滴鼻免疫小鼠2次,间隔14 d,以PBS 20μl/只滴鼻为对照。逐日观察小鼠状况,于末次免疫后第14天颈椎脱位处死小鼠,检测血清特异性IgG、小肠冲洗液sIgA水平,分离肠上皮内淋巴细胞(intesti-nal intraepithelial lymphocytes,iIEL)及肠系膜淋巴结淋巴细胞(mesenteric lymph node lymphocytes,MLNL)并计数。结果60 h组小鼠初次免疫后第8-11天,有倦怠、饮水及采食量减少等表现,体重增长较其他组缓慢(P<0.05),其他组小鼠健康状况良好。不同时相ESA鼻内免疫小鼠后特异性抗体水平升高,MLNLi、IEL发生增殖性应答。48 h组和60 h组各项指标均明显高于0 h组(P<0.01)及PBS组(P<0.05)。结论48 h和60 h弓形虫ESA均能诱导黏膜及系统免疫应答,但60h ESA可能对机体有毒副作用,不适宜直接滴鼻免疫,48 h ESA可作为弓形虫黏膜疫苗的候选抗原。　Objective To investigate immune responses after intranasal immunization with Toxoplasma gondii RH strain excreted-secreted antigens（ESA） in BALB/c mice.Methods ESA were taken from the infected murine peritoneal fluids after infection for 0 h,6 h,12 h,24 h,36 h,48 h and 60 h.Sixty-four BALB/c mice were randomly divided into 8 groups： experimental groups and control group.The mice in experimental groups were immunized intranasally with 20 μg ESA taken from the peritoneal fluids in infected mice at hours 0,6,12,24,36,48 and 60,respectively,while mice in control group were immunized with 20 μl PBS.All mice were twice immunized intranasally at interval of 14 d.The healthy status and body weight（BW） of mice were recorded every day.All mice were killed at day 14 after the last immunization.IgG in serum and sIgA in intestinal washes were detected by ELISA.Intestinal intraepithelial lymphocytes（iIEL）,mesenteric lymph node lymphocytes（MLNL） and spleen lymphocytes were isolated and counted. Results All mice were healthy and BW increased time-dependently,but the increase of BW in 60 h group was slower than in other groups（P〈0.05）.After intranasal immunization with ESA,level of specific antibodies in serum and mucosal site increased,and cell-proliferative responses were observed in iIEL,MLNL and spleen lymphocytes.The level of IgG in serum and sIgA in intestinal washes,the numbers of iIEL,MLNL in 48 h and 60 h groups were higher than in other groups（P〈0.05）.Conclusion The 48 h and 60 h ESA can effectively induce the mucosal and systemic immune responses.Thus 48 h ESA may be an attractive candidate antigen for Toxoplasma gondii vaccine,but 60 h ESA has toxicity and side effects in mice and could not be used in intranasal immunization.

关 键 词：弓形虫 免疫 黏膜 排泄-分泌抗原 鼻内免疫 

分 类 号：R382.5[医药卫生—医学寄生虫学]