摘要目的 探讨日本血吸虫重组双歧杆菌属两歧双歧杆菌(Bifidobacterium 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.

abstractsObjective 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.