铜绿假单胞菌重组Ef-PopB疫苗诱导小鼠脾细胞因子和Treg变化的研究  

Study on the changes in splenocyte cytokines and Treg in mice immunized with a recombinant Ef-PopB vaccine against Pseudomonas aeruginosa

作　　者：何爱琳 李文桂[1] HE Ai-lin;LI Wen-gui(Institute of Infections and Parasitic Diseases,the First Affiliated Hospital,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学附属第一医院传染病寄生虫病研究所,重庆400016

出　　处：《中国病原生物学杂志》2021年第3期266-269,共4页Journal of Pathogen Biology

基　　金：重庆市科委地方病专项基金项目(No.2008AB5055,2008AB5008,2008AB5054)。

摘　　要：目的观察铜绿假单胞菌重组Ef-PopB疫苗免疫及PA01株攻击后小鼠产生的保护力、脾细胞因子和Treg的变化。方法用5×10^(8) CFU的Ef-PopB疫苗灌胃免疫BALB/c鼠,每日接种1次,持续3 d,连续3周。初次免疫后4周,取5×107 CFU的PA01株滴鼻攻击小鼠。攻击后2周杀鼠,分离肺和脾,取肺组织培养肺细菌并行菌落计数;用PaAg培养脾细胞,PCR扩增IL-2、IL-4、IFN-γ、IL-10、IL-12和Foxp3基因。结果Ef-PopB疫苗组、空载体组和Ef对照组肺组织的菌落数分别为(0.37±0.02)×10^(8) CFU、(7.58±0.20)×10^(8) CFU和(7.55±0.19)×10^(8) CFU,差异有统计学意义(P<0.01);从疫苗组中抽提的脾细胞基因组的DNA分别扩增出453 bp的IL-2基因、359 bp的IL-4基因、399 bp的IFN-γ基因、301 bp的IL-10基因、300 bp的IL-12基因和250 bp的Foxp3基因。结论重组Ef-PopB疫苗可诱导小鼠脾细胞因子表达升高,产生混合型的Th1和Th2免疫应答。Objective To observe changes in the protective action and splenocyte cytokines and Treg in mice immunized with a recombinant Ef-PopB vaccine against Pseudomonas aeruginosa and challenged with the PA01 strain.Methods BALB/c mice received a daily intragastric administration of 5×10^(8) CFU of the Ef-PopB vaccine for 3 consecutive days.Intragastric administration was performed for a total of 3 weeks.Four weeks after the first immunization,mice were challenged intranasally with nasal drops of 5×107 CFU of the PA01 strain.Two weeks after the challenge,mice were sacrificed to collect the lungs and spleen.Bacteria in lung tissues were incubated and colonies were counted.Splenocytes were incubated with P.aeruginosa antigens,and the IL-2,IL-4,IFN-γ,IL-10,IL-12,and Foxp3 genes were amplified using PCR.Results The number of colonies in lung tissue was 0.37±0.02×10^(8) CFU in the Ef-PopB vaccine group,7.58±0.20×10^(8) CFU in the blank vector group,and 7.55±0.19×10^(8) CFU in the E.faecium control group.The number of colonies differed significantly among the groups(P<0.01).The IL-2 gene(453 bp in length),the IL-4 gene(359 bp in length),the IFN-γgene(399 bp in length),the IL-10 gene(301 bp in length),the IL-12 gene(300 bp in length),and the Foxp3 gene(250 bp in length)were separately amplified from the DNA of splenocytes from the vaccine group.Conclusion The recombinant Ef-PopB vaccine may induce increased expression of splenocyte cytokines in mice and produce mixed Th1 and Th2 immune responses.

关 键 词：铜绿假单胞菌 屎肠球菌 疫苗 细胞因子 

分 类 号：R378.991[医药卫生—病原生物学]