E.tenella河北株pcDNA3.0-IL-2-EtMIC-2真核表达载体的免疫调节作用
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摘要
为了研究柔嫩艾美耳球虫(E.tenella)河北株真核表达载体pcDNA3.0-IL-2-EtMIC-2的免疫调节作用,将240只14日龄雏鸡随机分成8组,第1,2组分别为阳性、阴性对照,在14,21 d,3、4组分别肌注100μg的pcDNA3.0-IL-2、pcDNA3.0-EtMIC-2;5、6、7组分别肌注50,100,150μg的pcDNA3.0-IL-2-EtMIC-2;8组21 d时滴口免疫鸡球虫病四价活疫苗。28 d,阳性对照组和6个试验组每只鸡经口攻毒4×10~4个E.tenella卵囊,阴性对照组每只鸡灌服同体积的生理盐水。结果显示:当pcDNA3.0-IL-2-EtMIC-2真核表达载体的免疫剂量达100,150μg/只时,试验后7,21,28 d IgG的含量、7 d的IgA含量和21,28 d的IFN-γ含量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);试验后28 d,血清中IL-2的含量显著高于阴性和阳性对照组(P<0.05),与单独免疫pcDNA3.0-IL-2和pcDNA3.0-EtMIC-2组相比差异不显著(P>0.05);试验后7,14,21,28 d脾脏IL-2和IFN-γmRNA表达量均显著高于阳性对照组、单独免疫pcDNA3.0-IL-2和pcDNA3.0-EtMIC-2组(P<0.05)。综上所述,IL-2可增强EtMIC-2基因的免疫原性,适量的真核表达载体pcDNA3.0-IL-2-EtMIC-2可显著提高感染柔嫩艾美尔球虫雏鸡的免疫功能。
To study the immunoregulatory effects of eukaryotic expression vector pcDNA3.0-IL-2-EtMIC-2 of E.tenella Hebei strain,240 14-day-old chicks were randomly divided into 8 groups.The first and second groups were positive and negative controls respectively.At 14 and 21 days,groups 3 and 4 were intramuscularly injected with 100 μg of pcDNA3.0-IL-2 and pcDNA3.0.-EtMIC-2,5,6,and 7 groups were intramuscularly injected with 50,100,and 150 μg of pcDNA3.0-IL-2-EtMIC-2,and 8 groups of mice were immunized with a four-valent live vaccine for oral coccidiosis.On the 28 th day,each chicken in the positive control group and the 6 test groups was challenged with 4×104 E.tenella oocysts by mouth,and each chicken in the negative control group was given the same volume of normal saline.The results showed that when the immunization dose of pcDNA3.0-IL-2-EtMIC-2 eukaryotic expression vector reached 100,150μg/head,the serum of IgG at 7,21,28 d,the content of IgA at 7 d,the content of IFN-γat 21,28 d,were significantly increased compared with the negative control group,positive control group,pcDNA3.0-IL-2 group and pcDNA3.0-EtMIC-2 group(P<0.05),At 28 days after the test,the serum IL-2 content was significantly higher than that of the negative and positive control groups(P<0.05),and the difference was not significant compared with the immunized pcDNA3.0-IL-2 and pcDNA3.0-EtMIC-2 groups alone(P>0.05);At 7,21,28 d,the expressions of IL-2 and IFN-γmRNA in spleen were significantly higher than those in the positive control group and pcDNA3.0-IL-2 and pcDNA3.0-EtMIC-2 groups(P <0.05).In summary,IL-2 can enhance the immunogenicity of EtMIC-2 gene,and the appropriate amount of eukaryotic expression vector pcDNA3.0-IL-2-EtMIC-2 can significantly improve the immune function of chicks infected with E.tenella.
To study the immunoregulatory effects of eukaryotic expression vector pcDNA3.0-IL-2-EtMIC-2 of E.tenella Hebei strain,240 14-day-old chicks were randomly divided into 8 groups.The first and second groups were positive and negative controls respectively.At 14 and 21 days,groups 3 and 4 were intramuscularly injected with 100 μg of pcDNA3.0-IL-2 and pcDNA3.0.-EtMIC-2,5,6,and 7 groups were intramuscularly injected with 50,100,and 150 μg of pcDNA3.0-IL-2-EtMIC-2,and 8 groups of mice were immunized with a four-valent live vaccine for oral coccidiosis.On the 28 th day,each chicken in the positive control group and the 6 test groups was challenged with 4×104 E.tenella oocysts by mouth,and each chicken in the negative control group was given the same volume of normal saline.The results showed that when the immunization dose of pcDNA3.0-IL-2-EtMIC-2 eukaryotic expression vector reached 100,150μg/head,the serum of IgG at 7,21,28 d,the content of IgA at 7 d,the content of IFN-γat 21,28 d,were significantly increased compared with the negative control group,positive control group,pcDNA3.0-IL-2 group and pcDNA3.0-EtMIC-2 group(P<0.05),At 28 days after the test,the serum IL-2 content was significantly higher than that of the negative and positive control groups(P<0.05),and the difference was not significant compared with the immunized pcDNA3.0-IL-2 and pcDNA3.0-EtMIC-2 groups alone(P>0.05);At 7,21,28 d,the expressions of IL-2 and IFN-γmRNA in spleen were significantly higher than those in the positive control group and pcDNA3.0-IL-2 and pcDNA3.0-EtMIC-2 groups(P <0.05).In summary,IL-2 can enhance the immunogenicity of EtMIC-2 gene,and the appropriate amount of eukaryotic expression vector pcDNA3.0-IL-2-EtMIC-2 can significantly improve the immune function of chicks infected with E.tenella.
引文
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[14] SHAH M A,YAN R,XU L,et al.A recombinant DNA vaccine encoding Eimeria acervulina cSZ-2 induces immunity against experimental E.tenella infection[J].Vet Parasitol,2010,169(1/2):185-189.
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[17] 刘颖丽,李建华,郑君,等.鸡柔嫩艾美耳球虫EtMIC-2基因真核表达载体的构建及其在Hela细胞中的表达[J].中国兽医学报,2011,31(8):1142-1146.
[2] SHIRLEY M W.Control of coccidiosis with living vaccines[J].World Poultry,1993(8/9):24-27.
[3] 张香斋,张召兴,李蕴玉,等.鸡球虫基因工程疫苗的研究进展(英文)[J].Agr Sci Tech J,2017,18(8):1480-1483.
[4] JENKINS M C,CASTLE M D,DANFORTH H D,Protective immunization against the intestinal parasite Eimeria acervulina with recombinant merozoite antigen[J].Immunology,1990,71:127-132.
[5] 李蕴玉,张召兴,李佩国,等.E.tenella河北株重组IL-2-EtMIC-2DNA疫苗免疫原性及抗氧化能力[J].中国兽医学报,2018,38(5):911-916.
[6] 张召兴,张香斋,李蕴玉,等.E.tenella河北株重组IL-2-EtMIC-2 DNA疫苗的构建及保护力分析[J].中国兽医学报,2018,38(1):77-81.
[7] RATCLIFFE M J.Antibodies,immunoglobulin genes and the bursa of Favricius in chicken B cell development[J].Dev Comp Immunol,2006,30(1/2):101-118.
[8] 张召兴.复方中草药超微粉防治鸡致病性大肠杆菌病的研究[D].河北秦皇岛:河北科技师范学院,2017.
[9] 李蕴玉,李佩国,张艳英,等.白细胞介素-2在兽医中的应用进展[J].河北科技师范学院学报,2006,20(4):63-66
[10] 张怀,高春生,蒋媛媛,等.重组白细胞介素-2对固始鸡免疫机能的影响[J].中国畜牧兽医,2009,36(7):48-54.
[11] 张伟,朱连勤,马凤龙,等鸡γ-干扰素的研究进展[J].畜牧兽医杂志,2007,26(2):43-44.
[12] SHAH M A,SONG X,XU L,et al.Construction of DNA vaccines encoding Eimeria acervulina cSZ-2 with chicken IL-2 and IFN-γ and their efficacy against poultry coccidiosis[J].Res Vet Sci,2011,90(1):72-77.
[13] 张娜.鸡毒害艾美耳球虫NA4基因免疫调节型DNA疫苗的构建及其保护性研究[D].江苏南京:南京农业大学,2008.
[14] SHAH M A,YAN R,XU L,et al.A recombinant DNA vaccine encoding Eimeria acervulina cSZ-2 induces immunity against experimental E.tenella infection[J].Vet Parasitol,2010,169(1/2):185-189.
[15] SONG H,YAN R,XU L,et al.Efficacy of DNA vaccines carrying Eimeria acervulina lactate dehydrogenase antigen gene against coccidiosis[J].Exp Parasitol,2010,126(2):224-231.
[16] GERILETU,XU L X,XU R H,et al.Vaccination of chickens with DNA vaccine expressing Eimeria tenella MZ5-7 against coccidiosis[J].Vet Parasitol,2011,177(1/2):6-12.
[17] 刘颖丽,李建华,郑君,等.鸡柔嫩艾美耳球虫EtMIC-2基因真核表达载体的构建及其在Hela细胞中的表达[J].中国兽医学报,2011,31(8):1142-1146.