日本血吸虫重组双歧杆菌属两歧双歧杆菌pGEX—Sj14—3—3疫苗免疫BALB／c小鼠脾细胞凋亡的动态观察  

Dynamic observation of splenocyte apoptosis in mice immunized with recombinant vaccine Bifulobacterium bifidum pGEX-Sj14-3-3 of Schistosoma japonicum

作　　者：张宁[1] 李文桂[2] 

机构地区：[1]重庆三峡中心医院综合医疗科,404000 [2]重庆医科大学附属第一医院传染病寄生虫病研究所寄生虫病研究室,重庆400016

出　　处：《中国地方病学杂志》2012年第6期604-607,共4页Chinese Jouranl of Endemiology

基　　金：重庆市科委地方病重大专项基金（2008AB5055、2008AB5008、2008AB5054）

摘　　要：目的探讨日本血吸虫重组双歧杆菌属两歧双歧杆菌（Bifutobacterium bifidum，Bb）pGEX—Sj14—3—3疫苗对BALB／c小鼠脾细胞凋亡的影响。方法96只BALB／c小鼠按体质量分为2组，每组48只，用重组Bb（pGEX—Sj14—3—3）疫苗分别经口服灌胃和鼻腔黏膜两种途径免疫小鼠，在免疫后0、2、4、6、8、10、12、14、16、18、20和22周各剖杀4只小鼠，取脾并分离脾细胞。两种免疫途径均分别用或不用刀豆蛋白A（ConA）刺激培养脾细胞，采用流式细胞仪检测脾细胞的凋亡发生率。结果未刺激时，口服灌胃（PO）组和鼻腔黏膜（IN）组小鼠的脾淋巴细胞凋亡水平在免疫后2～4周（PO组：0．069±0．005、0．076±0．010；IN组：0．037±0．002、0．075±0．002）显著升高，在免疫后4周达峰值（P均〈0．05）；ConA刺激时，两组小鼠的脾淋巴细胞凋亡水平在免疫后2—6周（PO组：0．089±0．006、0．098±0．010、0．060±0．007；IN组：0．054±0．001、0．093±0．003、0．058±0．012）升高，在4周达峰值（P均〈0．05）；每组ConA刺激时脾细胞凋亡发生率均明显高于相应的原液组（P均〈0．05）。结论重组Bb（pGEX—Sj14—3—3）疫苗口服及鼻腔黏膜免疫小鼠可抑制脾细胞发生凋亡。Objective To investigate the effects of recombinant vaccine Bifidobacterium bifidum （Bb） pGEX-Sj14-3-3 on splenocyte apoptosis in BALB/c mice. Methods Ninety-six BALB/c mice were randomly divided into two groups according to their body mass： per os group （PO） and intranasal immunization group （IN）, with 48 mice in each group. All mice were orally and intranasally immunized with recombinant vaccine Bb （pGEX-Sj14-3-3）. Four mice in each group were sacrificed on weeks 0, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20 and 22, respectively, after immunization, and splenocytes were separated and cultured with or without ConA stimulation. The apoptotic rates of splenocytes were detected by flow cytometry. Results It showed that apoptotic level of splenocytes in both groups remarkably increased after 2 - 4 weeks without ConA stimulation （PO： 0.069 ± 0.005, 0.076 ± 0.010; IN： 0.037 ± 0.002, 0.075 ± 0.002）, and the value reached the peak on the 4th week, and the differences were statistically significant compared with that of week 0（all P 〈 0.05）. Apoptotic level of splenocytes in both groups with ConA stimulation increased after 2- 6 weeks（PO： 0.089 ± 0.006, 0.098 ± 0.010, 0.060 ± 0.007; IN： 0.054 ± 0.001, 0.093 ± 0.003, 0.058 ± 0.012）, and the value also reached the peak after 4 week, respectively. The differences were statistically significant compared with that of week 0 （all P 〈 0.05 ）. Apoptotic level of splenocytes in each group with ConA stimulation was significantly higher than that without ConA stimulation. Conclusion It is suspected that the recombinant vaccine Bb（pGEX-Sj14-3-3） may inhibit apoptosis of splenocytes in mice immunized orally or intranasally.

关 键 词：血吸虫 日本 两歧双歧杆菌 疫苗 细胞凋亡 质粒 

分 类 号：R392[医药卫生—免疫学]