■犬瘟热(CD)是一种全球传播的疾病,已在12个哺乳动物家族中被描述,尤其是在食肉顺序中,在接种疫苗是最好的控制手段的国内犬类中进行更好的研究。CD通过疫苗接种控制,但许多病例的疾病仍然发生在接种疫苗的动物。
■这项工作的目的是研究可以资助新疫苗方法开发的抗原特异性表位。
■使用来自病毒血凝素蛋白的119个重叠合成肽,通过酶联免疫斑点测定法进行CD病毒(CDV)的T细胞反应性表位的定位,分为22个池,形成一个矩阵来测试32只动物的免疫反应。
■使用为识别反应池而建立的标准进行评估,证明26只动物至少有一个反应池,一个水池对任何动物都没有反应,和六个池是最常见的反应性肽。基质中最具反应性的池的交叉显示了9种肽,这些肽被认为是针对CDV的T细胞刺激的潜在候选表位,并用于设计计算机蛋白质。还含有B细胞刺激的预测表位,并使用免疫表位数据库进行进一步分析,以确保蛋白质质量和稳定性。
■最终的计算机模拟优化的蛋白质呈现有资格用于开发新的原型基于表位的抗CDV疫苗的特征。
UNASSIGNED: Canine distemper (CD) is a worldwide spread disease that has been described in 12 families of mammals, especially in the Carnivora order, being better studied in domestic canines where vaccination represents the best means of control. CD is controlled by vaccination, but many cases of the disease still occur in vaccinated animals.
UNASSIGNED: The aim of this work was to study antigen-specific epitopes that can subsidize the development of a new vaccine approach.
UNASSIGNED: Mapping of T cell reactive epitopes for CD virus (CDV) was carried out through enzyme-linked immunospot assays using 119 overlapped synthetic peptides from the viral hemagglutinin protein, grouped in 22 pools forming a matrix to test the immune response of 32 animals.
UNASSIGNED: Evaluations using the criteria established to identify reactive pools, demonstrated that 26 animals presented at least one reactive pool, that one pool was not reactive to any animal, and six pools were the most frequent among the reactive peptides. The crisscrossing of the most reactive pools in the matrix revealed nine peptides considered potential candidate epitopes for T cell stimulation against the CDV and those were used to design an in-silico protein, containing also predicted epitopes for B cell stimulation, and further analyzed using immune epitope databases to ensure protein quality and stability.
UNASSIGNED: The final in silico optimized protein presents characteristics that qualify it to be used to develop a new prototype epitope-based anti-CDV vaccine.