PRRSV灭活抗原免疫接种后抗主要结构蛋白抗体的产生规律研究  

Generation rules of anti-main structural protein antibodies induced by inactivated PRRSV antigen

作　　者：段二珍[1] 赵琳[1] 陈静[1] 张宜娜[1] 卢晓艳[1] 张志远[1] 刘肖萍[1] 夏平安[1] 张红英[1] 崔保安[1] 

机构地区：[1]河南农业大学牧医工程学院,郑州45000

出　　处：《免疫学杂志》2011年第3期234-238,共5页Immunological Journal

基　　金：国家自然科学基金(31040080)

摘　　要：目的检测PRRSV灭活抗原免疫接种后抗主要结构蛋白抗体的产生规律。方法用差速离心法纯化PRRSV Hn-3/06株,0.1%甲醛灭活PRRSV,加入弗氏佐剂乳化制备成PRRSV灭活抗原。20只昆明系小鼠(7～8周龄)随机分为2组,每组10只,每次每只110μg蛋白剂量,第1组免疫1次,第2组免疫2次,每次间隔7 d。每隔7 d采血1次,间接ELISA方法检测血清中抗PRRSV及GP5、M和N蛋白的抗体效价。用Marc-145细胞进行细胞中和试验,间接免疫荧光灶抑制方法检测血清中和抗体效价。结果 2组小鼠均产生针对PRRSV及各结构蛋白的抗体,抗PRRSV的抗体和抗N蛋白的抗体一免后14 d出现,21 d后抗体效价达到1∶1 280,35 d抗体水平达到最高;抗M蛋白的抗体一免后42 d出现,49 d抗体水平达到最高,抗体效价达到1:160;抗GP5蛋白的抗体一免后49 d出现,至56 d抗体水平呈上升趋势,抗体效价达到1∶80。免疫2次的小鼠各项ELISA抗体效价总体略微偏高。细胞中和试验结果表明第1组免疫小鼠免疫后42 d左右产生微弱的中和抗体;第2组免疫小鼠一免后35～49 d产生可检测到的中和抗体,42 d时中和抗体水平达到最高,抗体效价达到1∶2。结论纯化灭活的PRRSV抗原诱导机体产生的抗主要结构蛋白的抗体滴度较低,说明PRRSV抗原的免疫原性较弱,不能诱导机体产生有效的保护抗体。We aimed to detect generation rules of PRRSV main structural proteins specific antibodies induced by inactivated PRRSV antigen.PRRSV strain Hn-3/06 was purified by differential centrifugation,inactivated with formaldehyde,and emulsified with equal amount of Freund＇s adjuvant.Total of 20 mice（7~8 weeks old） were randomly assigned to two groups（10 mice each group）,and each mouse received 110 μg PRRSV antigen in Freund＇s adjuvant every time immunization.The group 1 was vaccinated once,while the group 2 was vaccinated twice,in 7 days interval.We collected serum weekly after vaccination,and detected PRRSV,GP5,M and N specific serum antibody titers by indirect ELISA.The results show that inactivated PRRSV antigen could induce PRRSV,GP5,M and N specific antibody in both experimental groups.Anti-PRRSV and N protein antibodies emergenced at 14 days post the primo vaccination,and the titers reached 1：1280 at 21 days,reached the highest level at 35 days post the primo immunization;anti-M protein antibodies appeared at 42 days post the primo vaccination,and the titers reached the highest level of 1∶160 at 49 days post the primo immunization;anti-GP5 protein antibodies appeared at 49 days post the primo vaccination and the antibody level was upgrading continuosly till 56 days,then the antibody titers reached 1∶80.Specific serum antibody titers of group 2 were totally slightly higher than those of group 2.Iindirect immunofluorescence focus inhibition test indicated that virus-neutralizing antibodies appeared in low amounts around 42 days post vaccination in group 1,while appeared between 35~49 days post the primo vaccination in group 2 and the antibody titers reached the highest level of 1∶2 at 42 days.In conclusion,the immunogenicity of PRRSV is weak and inactivated PRRSV antigen can not induce effective protection.

关 键 词：PRRSV ELISA 间接免疫荧光灶抑制试验 结构蛋白 免疫原性 

分 类 号：R392.11[医药卫生—免疫学]