羊种布鲁氏菌Rev.1突变株在BALB/c鼠中的免疫保护评价
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摘要
为评价Rev.1-ΔKan-Vir B12突变株在BALB/c鼠中的免疫保护力,以Rev.1为参照,用Rev.1-ΔKanVir B12接种BALB/c小鼠,免疫45 d后用布鲁氏菌16M强毒株攻毒,15 d后取BALB/c鼠脾脏进行克脾指数测定和病理组织学检测。结果显示:BALB/c鼠免疫攻毒15 d后,Rev.1-ΔKan-Vir B12免疫组和Rev.1免疫组的克脾指数与对照组之间有显著性差异(P<0.05),而Rev.1免疫组与Rev.1-ΔKan-Vir B12免疫组之间的克脾指数差异不显著(P>0.05)。结果表明,羊布鲁氏菌Rev.1-ΔKan-Vir B12突变株与其亲本Rev.1株的免疫保护性无明显差异,具备作为布鲁氏菌病标记疫苗的潜力。
In order to evaluate protective efficacy of Rev.1-ΔKan-VirB12 mutant in BALB/c mice,BALB/c mice were vaccinated with both Rev.1 and Rev.1-ΔKan-VirB12,and then were challenged with 16 M after vaccinated 45 d. Fifteen days post challenge,the spleens of BALB/c mice were taken for the Brucella load of gram of spleen determination and histopathological examination. Results showed that there was significant difference between Rev.1-ΔKan-VirB12 group and Rev.1 group and control group(P<0.05). The difference of Brucella load of gram of spleen between Rev.1 immune group and Kan-Vir B12 immune group was not significant(P>0.05). The results showed that the protective efficacy of both was nearly same,holding the potential as a vaccine for Brucellosis.
In order to evaluate protective efficacy of Rev.1-ΔKan-VirB12 mutant in BALB/c mice,BALB/c mice were vaccinated with both Rev.1 and Rev.1-ΔKan-VirB12,and then were challenged with 16 M after vaccinated 45 d. Fifteen days post challenge,the spleens of BALB/c mice were taken for the Brucella load of gram of spleen determination and histopathological examination. Results showed that there was significant difference between Rev.1-ΔKan-VirB12 group and Rev.1 group and control group(P<0.05). The difference of Brucella load of gram of spleen between Rev.1 immune group and Kan-Vir B12 immune group was not significant(P>0.05). The results showed that the protective efficacy of both was nearly same,holding the potential as a vaccine for Brucellosis.
引文
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[9]易德武,张俊波,王远志,等.羊种布鲁氏菌新疆流行株015株sodc基因缺失株的构建与初步评价[J].石河子大学学报(自然科学版),2016,34(1):24-29.
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[13]李默,易德武,王远志,等.布鲁氏菌Ⅳ型分泌系统效应蛋白Bsp B的原核表达及布鲁氏菌免疫逃逸分析[J].中国病原生物学杂志,2015,10(9):778-781.
[14]O'CALLAGHAN D,CAZEVIEILLE C,ALLARDETSERVENT A,et al.A homologue of the Agrobacterium tumefaciens Vir B and Bordetella pertussis Ptl type IV secretion systems is essentialfor intracellular survival of Brucellasuis[J].Molecular Microbiology,1999,33(6):1210-1220.
[15]YAO H S,HORTENSIA G R,ANDREAS B,et al.Brucella abortus Vir B12 is expressed during infection but is not an essential component of the type IV secretion system[J].Infectionand Immunity,2005,73(9):6048-6054.
[16]丁家波,王芳,杨宏军,等.一株中等毒力牛种布鲁氏菌的鉴定和毒力测定[J].中国农业科学,2014,47(13):2652-2658.
[17]易新萍,谷文喜,李金平,等.评价牛型布鲁氏菌疫苗免疫保护力BALB/c鼠模型的建立[J].中国兽医杂志,2013,40(6):73-77.
[18]郑孝辉,胡森,王加兰,等.流产布氏杆菌S19突变株构建及在小鼠感染模型中的免疫保护评估[J].中国预防兽医学报,2009,31(11):887-891.
[2]马晓菁,库尔班·居麦,舒展,等.2012-2014年新疆动物布鲁氏菌病流行病学调查与分析[J].畜牧与兽医,2016,48(5):111-114.
[3]GALINDO R C,MUNOZ P M,MIGUEL M J,et al.Genecxpression changes in spleens of wildlife reservoir species,Eurasian wild boar(Susscrofa),naturally infected with Brucella suisbiovar 2[J].Journal of Genetics and Genomics,2010,37(11):725-736.
[4]毛开荣.布鲁氏菌病研究进展[J].动物保健,2006,109(9):28-30.
[5]WYCKOFF J H,POTTS R D.Killing of Brucella antigen-sensitized macrophages by T lymphocytes in bovine brucellosis[J].Veterinary Immunology and Immunopathology,2007,120(3/4):148-159.
[6]JACQUES I,VERGER J M,LAROUCAU K,et al.Immunological responses and protective efficacy against Brucella melitensis induced by bp26 and omp31 B.melitensis Rev.1 deletion mutants in sheep[J].Vaccine,2007,25(5):794-805.
[7]CLOECKAET A,JACQUES I,GRILLOM J,et al.Development and evaluation as vaccines in mice of Brucella melitensis Rev.1 single and double deletion mutants of the BP26and omp31 genes coding for antigens of diagnostic significance in ovine brucellosis[J].Vaccine,2004,22:2827-2835.
[8]钟旗,何倩倪,范伟兴,等.布鲁氏菌Vir B8变异株的构建及其感染力和毒力的测定[J].畜牧兽医学报,2009,40(6):892-897.
[9]易德武,张俊波,王远志,等.羊种布鲁氏菌新疆流行株015株sodc基因缺失株的构建与初步评价[J].石河子大学学报(自然科学版),2016,34(1):24-29.
[10]CAMPOS E,CRAVERO S L,DELGUI L,et al.Brucella abortus INTA2,a novel strain 19Δbp26:lucΔbmp18 double mutant lacking drug resistance markers[J].Veterinary Microbiology,2002,87(1):1-13.
[11]ROLáNH G,DENHARTIGHA B,KAHLMCDONAGH M,et al.Vir B12 is a serological marker of Brucella infection in experimental andnatural hosts[J].Clinic Vaccine Immunology,2008,15(2):208-214.
[12]STEVENSM G,OLSENS C,PUGHG W,et al.Comparison of immune responses and resistance to Brucellosisin mice vacciuated with Brucella abortus 19 or RB51[J].Infection Immunity,1995,63(1):264-270.
[13]李默,易德武,王远志,等.布鲁氏菌Ⅳ型分泌系统效应蛋白Bsp B的原核表达及布鲁氏菌免疫逃逸分析[J].中国病原生物学杂志,2015,10(9):778-781.
[14]O'CALLAGHAN D,CAZEVIEILLE C,ALLARDETSERVENT A,et al.A homologue of the Agrobacterium tumefaciens Vir B and Bordetella pertussis Ptl type IV secretion systems is essentialfor intracellular survival of Brucellasuis[J].Molecular Microbiology,1999,33(6):1210-1220.
[15]YAO H S,HORTENSIA G R,ANDREAS B,et al.Brucella abortus Vir B12 is expressed during infection but is not an essential component of the type IV secretion system[J].Infectionand Immunity,2005,73(9):6048-6054.
[16]丁家波,王芳,杨宏军,等.一株中等毒力牛种布鲁氏菌的鉴定和毒力测定[J].中国农业科学,2014,47(13):2652-2658.
[17]易新萍,谷文喜,李金平,等.评价牛型布鲁氏菌疫苗免疫保护力BALB/c鼠模型的建立[J].中国兽医杂志,2013,40(6):73-77.
[18]郑孝辉,胡森,王加兰,等.流产布氏杆菌S19突变株构建及在小鼠感染模型中的免疫保护评估[J].中国预防兽医学报,2009,31(11):887-891.