摘要
为了探寻转移因子(Transfer factor,TF)对猪丹毒杆菌SpaA(Surface protective antigen A)蛋白抗原免疫效果的影响,将重组SpaA蛋白与双相佐剂乳化后(称SpaA抗原)免疫小鼠,取免疫小鼠和未免疫小鼠脾脏,以超滤法分别制备特异性TF和普通TF,并利用BCA法测定所得TF的质量分数,经过无菌及安全性检测后,将2种TF分别腹腔接种至小鼠体内,24 h后,对2组TF处理后的小鼠均免疫SpaA抗原,并以未接种TF的免疫小鼠作为对照(仅免疫组)。分别于免疫后12 d、21 d和30 d对小鼠进行采血,用间接ELISA法对小鼠血清进行抗体检测。结果表明,利用原核表达系统表达的重组SpaA蛋白分子质量60 ku且具有可溶性;以超滤法制备的特异性TF质量浓度为2.56 mg/mL,普通TF质量浓度为2.40 mg/mL,且2种TF安全无毒;抗体检测数据显示,2种TF处理组在免疫后21、30 d的抗体水平均高于仅免疫组,且差异显著,而2种TF处理组之间抗体水平差异不显著。因此,TF能诱导SpaA抗原快速产生抗体,提高SpaA抗原的免疫效果,但与TF的特异性关系不大,表明TF可作为猪丹毒杆菌基因工程亚单位疫苗的免疫增效剂,提高免疫动物的抗体水平。
The aim was to explore the effect of transfer factor on immune efficacy of SpaA antigen of Erysipelothrix rhusiopathiae.Mice were immunized with the recombinant SpaA antigen emulsified with two-phasic adjuvant.And then,the SpaA antigen-specific TF and normal TF were prepared from immunized and non-immunized mouse spleens by ultrafiltration.Both two kinds of TFs were injected into the mouse abdomen after the BCA concentration determination,sterility test and safty test.24 hours later,mice treated with TFs were immunized with SpaA antigen while the untreated mice were used as control group(only immunized group).Blood was collected at 12,21,30 d after the last vaccination and tested by indirect enzyme-linked immuno-sorbent assay.The results showed that the recombinant SpaA protein expressed by prokaryotic expression system was about 60 ku and soluble,the SpaA antigen-specific TF peptide content was 2.56 mg/mL and the normal TF was 2.40 mg/mL prepared by ultrafiltration method,and the two TFs were safe and non-toxic.Antibody detection data indicated that the two groups treated with TFs had a higher antibody level than the group of only immunized group at 21 and 30 days after immunization,which showed significant differences,while there were no significant differences between the two groups treated with TFs.Therefore,TFs could induce SpaA antigen to rapidly produce antibodies and improve immune efficacy.However,it has little relation with the specificity of TFs.It also indicates that TFs can be used as an immunopotentiator for the genetic engineering subunit vaccine of Erysipelothrix rhusiopathiae to increase the antibody level of immunized animals.
作者
胡辉灿
刘洋
姚清
赵墩
余兴龙
HU Huican;LIU Yang;YAO Qing;ZHAO Dun;YU Xinglong(College of Veterinary Medicine,Hunan Agriculture University,Changsha 410128,China)
出处
《河南农业科学》
北大核心
2019年第4期135-138,共4页
Journal of Henan Agricultural Sciences
基金
湖南省战略性新兴产业科技攻关类项目(2016GK4015)
关键词
转移因子
特异性
猪丹毒杆菌
SpaA抗原
抗体水平
Transfer factor
Specificity
Erysipelothrix rhusiopathiae
SpaA antigen
Antibody levels
