摘要
为了研究猪附红细胞体eno基因重组蛋白对仔猪的免疫效果,本研究将9头仔猪随机平均分为3组:免疫组A皮下多点注射免疫纯化的猪附红细胞体重组蛋白rMseno,剂量为2.5 mg/头;免疫组B皮下注射免疫诱导后的重组菌Mseno/E.coli裂解液,剂量为5×10^(8)cfu/头;对照组C接种等量PBS;共免疫2次,2次免疫间隔21 d。在一免后0、7 d、14 d、21 d、28 d、35 d采集仔猪血清,通过间接ELISA方法检测血清中重组蛋白特异性抗体、IgG_(1)、IgG_(2a)、细胞因子IFN-γ和IL-4水平;流式细胞术检测免疫仔猪CD4^(+)T细胞、CD8^(+)T细胞水平,评价重组蛋白对仔猪的免疫效果。间接ELISA方法检测结果显示,免疫组A和免疫组B猪血清中重组蛋白特异性抗体、IgG_(1)、IgG_(2a)、细胞因子IL-4和IFN-γ水平均极显著高于对照组C(p<0.01);流式细胞术检测结果显示,免疫组A和免疫组B CD4^(+)T细胞水平极显著高于对照组C(p<0.01);免疫组A和免疫组B的CD8+T细胞水平和CD4^(+)T细胞/CD8+T细胞比值显著高于对照组C(p<0.05);免疫组A与免疫组B比较,上述所测指标均差异不显著(p>0.05)。结果表明,猪附红细胞体eno基因重组蛋白能诱导仔猪产生较高的体液免疫和细胞免疫水平,纯化后的重组蛋白与表达重组蛋白的重组菌裂解液所诱导的免疫效果差异不显著。本研究首次证实猪附红细胞体eno基因重组蛋白对仔猪的免疫效果,为猪附红细胞体病亚单位疫苗的研发提供参考依据。
In order to investigate the immune effect of eno gene recombinant protein on piglets,nine piglets are randomly divided into three groups.Piglets in group A or group B were vaccinated with 2.5 mg purified eno gene recombinant protein of Mycoplasma suis or 5×10^(8) cfu induced recombinant bacteria Mseno/E.coli lysate subcutaneously on day 0 and again 21 days post immunization;piglets in group C received the same amount of PBS.To evaluate the immune effect of recombinant protein on piglets,the serum of piglets was collected at 0,7,14,21,28 and 35 days after the first immunization,and the levels of antirecombinant protein antibodies,IgG_(1),IgG_(2a),IFN-γand IL-4 were detected by indirect ELISA;The percentage CD4^(+)T cells and CD8^(+)T cells were detected by flow cytometry.The indirect ELISA results showed that the levels of anti-recombinant protein antibodies,IgG_(1),IgG_(2a),IL-4 and IFN-γin serum of group A and group B were significantly higher than those of control group(p<0.01);the flow cytometry results showed that the percentage of CD4^(+)T cells and CD8^(+)T cells in spleen of group A and group B were significantly higher than those of control group C(p<0.01);as well as the ratio of CD4^(+)/CD8^(+)T cells in group A and group B was significantly higher than that in control group C(p<0.05).There were no significant differences in the above data between the group A and group B(p>0.05).These results demonstrated that the eno gene recombinant protein of Eperythrozoon suis could induce good humoral and cellular immunity in piglets,and there was no significant difference between the purified recombinant protein and the lysate of recombinant bacteria expressing the recombinant protein.This is the first report about the immune effect of eno gene recombinant protein of of Eperythrozoon suis on piglets,and is meaningful for the development of subunit vaccine of Eperythrozoon suis.
作者
韩金成
伍生军
闫宗斌
于啸洋
金鑫
李文学
许应天
薛书江
HAN Jin-cheng;WU Sheng-jun;YAN Zong-bin;YU Xiao-yang;JIN Xin;LI Wen-xue;XU Ying-tian;XUE Shu-jiang(Agricutural College in Yanbian University,Yanji 133002,China;Engineering Research Center of North-East Cold Region Beef Cattle Science&Technology Innovation,Ministry of Education,Yanbian University,Yanji 133002,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2021年第11期1192-1196,1218,共6页
Chinese Journal of Preventive Veterinary Medicine
基金
国家自然科学基金(31460657)
吉林省教育厅“十三五”科学技术项目(JJKH20191134KJ)
高等学校学科创新引智计划(D20034)
