E.tenella河北株重组IL-2-EtMIC-2DNA疫苗免疫原性及抗氧化能力
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摘要
为了确定柔嫩艾美耳球虫(E.tenella)河北株重组IL-2-EtMIC-2DNA疫苗的免疫原性与抗氧化能力,将240只14日龄雏鸡随机分成8组,每组30只,Ⅰ、Ⅱ组分别为阳性对照组和阴性对照组,Ⅲ、Ⅳ组在14、21日龄分别肌肉注射100μg的pcDNA3.0-IL-2、pcDNA3.0-EtMIC-2,Ⅴ、Ⅵ、Ⅶ组在14、21日龄分别肌肉注射50,100,150μg的pcDNA3.0-IL-2-EtMIC-2,Ⅷ组在21日龄免疫鸡球虫活疫苗;28日龄时,除Ⅱ组外,每只鸡接种4×10~4个E.tenella卵囊。结果显示:各免疫组鸡的外周血T淋巴细胞转化率、血清EtMIC-2抗体水平以及血清T-SOD、GSH-Px和TAOC活性较Ⅰ、Ⅱ组均呈现升高趋势,其中当pcDNA3.0-IL-2-EtMIC-2重组质粒的免疫剂量达100μg/只时,使试验21d的外周血T淋巴细胞转化率、14,21,28d的血清EtMIC-2抗体水平和T-AOC活性,14,21d的血清GSH-Px活性以及14d的血清T-SOD活性均显著高于阴性对照组、阳性对照组、单独免疫pcDNA3.0-IL-2和pcDNA3.0-EtMIC-2组(P<0.05)。结果表明:IL-2可增强EtMIC-2基因的免疫原性和抗氧化能力,适量的pcDNA3.0-IL-2-EtMIC-2重组质粒可显著提高机体的细胞与体液免疫水平以及抗氧化能力,达到抗球虫的效果。
In order to determine the immunogenicity and antioxidant capacity of recombinant IL-2-EtMIC-2 DNA vaccine,240 chickens at 14 days old were randomly divided into 8 groups of 30 each,including positive control group(groupl) and negative control group(group2), intramuscular injection pcDNA3.0-IL-2 and pcDNA3.0-EtMIC-2(100 μg/each bird) groups at 14,21 days old respectively(3,4 groups),intramuscular injection recombinant plasmid pcDNA3. 0-IL-2-EtMIC-2(50.100,150μg/each bird) groups at 14,21 days old,respectively(5,6,7 groups),oral live attenuated coccidiosis vaccine immunization group( 8 group) at 21 days old. All birds except the group 2 were inoculated with 4×10~4 sporulated E. tenella oocysts at 28 days.The results indicated that the T lymphocyte transformation rate of peripheral blood, serum EtMIC-2 antibody level,T-SOD,GSH-Px and T-AOC activity in all the immune groups showed an upward trend compared with the control groups, When the immunizing dose of recombinant plasmid pcDNA3.0-IL-2-EtMIC-2 was100 μg/each bird,the peripheral blood T lymphocyte transformation rate at 21,serum EtMIC-2 group,pcDNA3.0-IL-2 and pcDNA3.0-EtMIC-2 groups(P<0.05).The results indicated that IL-2 could enhance the immunogenicity and antioxidant activity of EtMIC-2 gene,and appropriate amount of recombinant plasmid pcDN A3. 0-IL-2-EtMIC-2 could significantly improve the cellular and humoral immunity and antioxidant capacity, resulting in the effect of anticoccidiosis.
In order to determine the immunogenicity and antioxidant capacity of recombinant IL-2-EtMIC-2 DNA vaccine,240 chickens at 14 days old were randomly divided into 8 groups of 30 each,including positive control group(groupl) and negative control group(group2), intramuscular injection pcDNA3.0-IL-2 and pcDNA3.0-EtMIC-2(100 μg/each bird) groups at 14,21 days old respectively(3,4 groups),intramuscular injection recombinant plasmid pcDNA3. 0-IL-2-EtMIC-2(50.100,150μg/each bird) groups at 14,21 days old,respectively(5,6,7 groups),oral live attenuated coccidiosis vaccine immunization group( 8 group) at 21 days old. All birds except the group 2 were inoculated with 4×10~4 sporulated E. tenella oocysts at 28 days.The results indicated that the T lymphocyte transformation rate of peripheral blood, serum EtMIC-2 antibody level,T-SOD,GSH-Px and T-AOC activity in all the immune groups showed an upward trend compared with the control groups, When the immunizing dose of recombinant plasmid pcDNA3.0-IL-2-EtMIC-2 was100 μg/each bird,the peripheral blood T lymphocyte transformation rate at 21,serum EtMIC-2 group,pcDNA3.0-IL-2 and pcDNA3.0-EtMIC-2 groups(P<0.05).The results indicated that IL-2 could enhance the immunogenicity and antioxidant activity of EtMIC-2 gene,and appropriate amount of recombinant plasmid pcDN A3. 0-IL-2-EtMIC-2 could significantly improve the cellular and humoral immunity and antioxidant capacity, resulting in the effect of anticoccidiosis.
引文
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[12]汪小红,武书庚,崔耀明,等.茶多酚对蛋鸡生产性能、蛋品质和抗氧化能力的影响[J].动物营养学报,2017,29(1):193-201.
[13]金光明,张岱芋,顾有方,等.柔嫩艾美耳球虫感染后组织抗氧化功能的动态变化[J].中国兽医学报,2002,22(3):233-235.
[14]聂奎,汤明,鲁智,等.柔嫩艾美耳球虫感染雏鸡LPO含量和抗自由基酶活性的动力学[J].中国兽医学报,2003,23(5):469-471.
[15]李佩国,李蕴玉,贾青辉,等.地克珠利对球虫感染雏鸡血清抗氧化功能的影响[J].中国家禽,2007,29(23):24-26.
[16]YANG W,WAINE G J,MCMANUS D P.Antibodies to schistosoma japonicum(Asian blood fluke)paramyosin induced by nucleic acids vaccination[J].Biochem Biophys Res Commun,1995,212(3):1029-1039.
[17]XU D,LIEW F Y.Genetic vaccination against leishmaniasis[J].Vaccine,12(16):1534-1536.
[18]SEDEGAH M,HEDSTROM R,HOBART P,et al.Protection against malaria by immunization with plasmid DNA encoding circumsporozoite protein[J].Proc Natl Acad Sci USA,1994,91(21):9866-9870.
[19]侯洪烈,张许科,李运娜,等.ADF-IL-2基因重组卡介苗的构建及免疫保护效果[J].中国兽医学报,2013,33(7):1016-1020.
[2]丁熙成,蒋金书.EtMIC-2表达产物对鸡3种艾美耳球虫的保护试验[J].畜牧兽医学报,2002,33(2):205-208.
[3]SATHISH K,SRIRAMAN R,SUBRAMANIAN B M,et al.Plant expressed EtMIC2is an effective immunogen in conferring protection against chicken coccidiosis[J].Vaccine,2011,29(49):9201-9208.
[4]DING X,LILLEHOJ H S,DALLOUL R A,et al.In ovo vaccination with the Eimeria tenella EtMIC-2gene induces protective immunity against coccidiosis[J].Vaccine,2005,23(28):3733-3740.
[5]MIN W,LILLEHOJ H S.Adjuvant effects of IL-1,beta,IL-2,IL-8,IL-15,INF-alpha,IFN-gamma,TGFbeta 4and lymphotactin on DNA vaccination against Eimeria acervulina[J].Vaccine,2001,20(122):267-274.
[6]SONG H,QIU B,YAN R,et al.The protective efficacy of chimeric SO7/IL-2DNA vaccine against coccidiosis in chickens[J].Res Vet Sci,2013,94(3):562-567.
[7]张杰.鸡柔嫩艾美尔球虫微线蛋白-2的真核表达及免疫原性研究[D].山东泰安:山东农业大学,2014.
[8]崔治中,朱瑞良.兽医免疫学实验指导[M].北京:中国农业大学出版社,2015.
[9]陈瑗,周玫.自由基与衰老[M].北京:人民卫生出版社,2004:10-20.
[10]郑荣梁,中洋.自由基生物学[M].北京:高等教育出版社,2007:9-24.
[11]董亮,何永志,王远亮,等.超氧化物歧化酶SOD的应用研究进展[J].中国农业科技导报,2013,15(5):53-58.
[12]汪小红,武书庚,崔耀明,等.茶多酚对蛋鸡生产性能、蛋品质和抗氧化能力的影响[J].动物营养学报,2017,29(1):193-201.
[13]金光明,张岱芋,顾有方,等.柔嫩艾美耳球虫感染后组织抗氧化功能的动态变化[J].中国兽医学报,2002,22(3):233-235.
[14]聂奎,汤明,鲁智,等.柔嫩艾美耳球虫感染雏鸡LPO含量和抗自由基酶活性的动力学[J].中国兽医学报,2003,23(5):469-471.
[15]李佩国,李蕴玉,贾青辉,等.地克珠利对球虫感染雏鸡血清抗氧化功能的影响[J].中国家禽,2007,29(23):24-26.
[16]YANG W,WAINE G J,MCMANUS D P.Antibodies to schistosoma japonicum(Asian blood fluke)paramyosin induced by nucleic acids vaccination[J].Biochem Biophys Res Commun,1995,212(3):1029-1039.
[17]XU D,LIEW F Y.Genetic vaccination against leishmaniasis[J].Vaccine,12(16):1534-1536.
[18]SEDEGAH M,HEDSTROM R,HOBART P,et al.Protection against malaria by immunization with plasmid DNA encoding circumsporozoite protein[J].Proc Natl Acad Sci USA,1994,91(21):9866-9870.
[19]侯洪烈,张许科,李运娜,等.ADF-IL-2基因重组卡介苗的构建及免疫保护效果[J].中国兽医学报,2013,33(7):1016-1020.