摘要
塞内卡病毒A(Senecavirus A)是小RNA病毒科塞内卡病毒属的唯一成员,仔猪感染后死亡率高达30%~70%,给全球养猪业造成了巨大的损失。开展抗原表位的研究对于病原学研究、免疫监测、疾病诊断及表位疫苗设计具有重要意义,细胞免疫作为清除胞内微生物最有效的防御手段,在免疫应答中起到了十分重要的作用。为了筛选塞内卡病毒A结构蛋白T细胞抗原表位,我们构建了真核表达质粒pcDNA3.1-P1,验证体外表达情况后,将其作为DNA疫苗免疫小鼠,通过流式细胞术检测免疫小鼠后刺激淋巴细胞增殖情况,通过酶联免疫斑点试验筛选SVA结构蛋白T细胞抗原表位,最终选择重复率高且效果较好的多肽,经检测,该多肽具有刺激淋巴细胞产生IFN-γ的能力。结果表明,通过对SVA结构蛋白的筛选,发现1号肽(SFDTASGTF)、9号肽(MPFQSLGTY)、11号肽(ITLTFCGPM)、21号肽(TSVDISVPYI)的效果较好,为塞内卡病毒A的T细胞抗原表位。
Senecavirus A is the only member of the Senecavirus genus of the Picornaviridae family.The mortality rate of piglets after infection ranges from 30% to 70%,causing huge losses to the global pig industry. The epitope research is of great significance to pathogenic research,immune monitoring,disease diagnosis,and epitope-based vaccine design. Cellular immunity,as the most effective defense method to eliminate intracellular microorganisms,plays a very important role in the immune response. To screen the T cell epitopes in the structural protein of Seneca virus A,we constructed the eukaryotic expression vector pc DNA3.1-P1. After verifying its expression in vitro,it was used as a DNA vaccine to immunize mice. The proliferation level of mouse lymphocytes after vaccination was detected by flow cytometry. The T cell epitopes in SVA structural protein were screened by enzyme-linked immunospot assay. Then the peptides with high repetition rate and good effect would be selected to test their ability to stimulate the lymphocytes to produce IFN-γ.The results showed that through the screening,there were four peptides with a better effect,which were named peptide No.1(SFDTASGTF),peptide No.9(MPFQSLGTY),peptide No.11(ITLTFCGPM),peptide No.21(TSVDISVPYI).Hence these peptides are the T cell epitopes of Senecavirus A.
作者
郭丽莹
冯霞
潘颂佳
马军武
GUO Li-ying;FENG Xia;PAN Song-jia;MA Jun-wu(Lanzhou Veterinary Research Institute,Chinese Academy of Agric ultural Sciences/State Key Laboratory of Veterinary Eiological Biolog/Nationad Foot and Mouth Disease Reference Laboratory,Lanzhou 730046,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2022年第1期77-84,共8页
Chinese Veterinary Science
基金
甘肃省自然科学基金项目(20JR5RA584):塞内卡病毒VP1蛋白B细胞表位和T细胞表位图谱的绘制。
