摘要
为评价猪圆环病毒2型病毒样颗粒(PCV2-VLP)和塞内卡病毒(SVV)全病毒灭活组合物的免疫效果。本研究首先通过透射电镜对PCV2-VLP的完整性进行鉴定,结果显示,PCV2-VLP颗粒完整,大小均一,直径为17 nm~20 nm。表明PCV2-VLP完整性良好,可以用于后续研究。然后将PCV2-VLP与SVV全病毒灭活病毒液等体积混合,制备二联组合物。将二联组合物、PCV2-VLP和SVV全病毒灭活病毒液分别以603和卡波姆作为佐剂两次免疫小鼠,并设立对照组。初免后每周采血,通过ELISA检测PCV2特异性抗体水平及IL-4和IFN-γ的分泌水平,通过中和试验检测小鼠血清针对PCV2和SVV的中和抗体。结果显示,相比卡波姆佐剂,二联组合物、PCV2-VLP和SVV全病毒灭活病毒液配合603佐剂免疫均能够诱导小鼠产生更高的特异性抗体及中和抗体。在相同佐剂下,二联组合物免疫小鼠产生的PCV2抗体与PCV2-VLP组相当,产生的SVV中和抗体则高于SVV单独免疫组(p<0.05)。各免疫组小鼠产生的细胞因子含量相当。表明,603佐剂优于卡波姆佐剂;二联组合物可以刺激小鼠产生针对PCV2和SVV的高水平中和抗体,具有良好的免疫原性。本研究首次将PCV2-VLP与SVV全病毒灭活病毒液进行联合免疫,在小鼠上证明具有良好的免疫原性,具有很大的参考价值,为进一步猪的免疫实验提供参考依据。
In order to evaluate of the immunogenicity of bivalent Porcine circovirus type 2 virus-like particles(PCV2-VLP)and inactivated Seneca valley virus(SVV) complex, integrity of PCV2-VLP particles were verified by transmission electron microscopy firstly. The results showed that PCV2-VLP particles are complete and uniform in size with a diameter of about 17 nm-20 nm, indicating that PCV2-VLP has good integrity and can be used for further vaccine research. The equal amounts of PCV2-VLP and SVV inactivated virus solution were mixed. Mice were immunized twice with complex, PCV2-VLP and SVV inactivated virus solution with 603 and carbomer as adjuvant respectively, and the control group was established meanwhile. After the initial immunization, antibodies and cytokines were examined by enzyme-linked immunosorbent assay(ELISA), and neutralizing antibodies were examined by neutralization test. The results showed that compared with carbomer adjuvant, complex, PCV2-VLP and SVV inactivated virus solution combined with 603 adjuvant could produce higher specific antibody and neutralizing antibody.With the same adjuvant, both mixed complex and PCV2-VLP could induce a comparable level of specific antibody and neutralizing antibody against PCV2, SVV neutralizing antibody produced by the bivalent mixed complex were significantly higher than those of in single group(p<0.05). In addition, the cytokine levels were the same in each group. These findings suggest that603 adjuvant was better than carbomer adjuvant;the complex was able to produce high level neutralizing antibody against PCV2 and SVV, and had good immunogenicity. In general, the bivalent PCV2-VLP and inactivated SVV complex was used for immunogenicity evaluation for the first time. It has been proved that the complex has good immunogenicity in mice shown great reference value, which provide reference basis for further pig immunity test.
作者
段卫同
王海伟
孙明霞
王尚辉
王刚
于力
蔡雪辉
涂亚斌
DUANWei-tong;WANG Hai-wei;SUN Ming-xia;WANG Shang-hui;WANG Gang;YU Li;CAI Xue-hui;TU Ya-bin(State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2021年第10期1080-1084,1107,共6页
Chinese Journal of Preventive Veterinary Medicine
基金
国家重点研发计划项目(2017YFD0500600)。
