Selection of T-cell epitopes from foot-and-mouth disease virus reflects the binding affinity to different cattle MHC class II molecules
Selection of T-cell epitopes from foot-and-mouth disease virus reflects the binding affinity to different cattle MHC class II molecules
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DOI：
10.1007/s002510000205
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发表时间：
2000-07-01
期刊：
影响因子：
3.2
通讯作者：
Hensen, EJ
中科院分区：
文献类型：
--
作者：
Haghparast, A;Wauben, MHM;Hensen, EJ
应用主要组织相容性复合体（MHC）肽段直接结合试验，研究了牛MHC Ⅱ类DR（BoLA-DR）分子对口蹄疫病毒（FMDV）T细胞表位的限制性选择。通过在体外引发的T淋巴细胞来源于动物的MHC I类和II类的纯合子，五个T细胞表位进行了分析的背景下，三个MHC II类单倍型。我们发现这些T细胞表位的呈递是由DR分子介导的，因为使用单克隆抗体TH 14 B阻断这种抗原呈递途径完全消除了对肽的增殖反应。为了研究这些T细胞表位的DR限制性呈递，开发了对分离的牛DR分子的直接MHC肽结合测定。从外周血单核细胞中分离纯化的BoLA-DRB 3 *0201、BoLA-DRB 3 *1101和BoLADRB 3 *1201等位基因的牛MHC II类DR分子。对于每个等位基因，将鉴定的T细胞表位之一生物素化，并用作标记肽用于开发竞争性MHC-肽结合测定。随后，在该结合测定中分析了具有功能上定义的MHC II类特异性的FMDV的T细胞表位。表位与某些DR分子结合的亲和力与诱导T细胞增殖的能力显著相关。这在分子水平上证明了在功能水平上发现的单个T细胞表位的选择确实是MHC限制的结果。
The major histocompatibility complex (MHC)-restricted selection of T-cell epitopes of foot-and-mouth disease virus (FMDV) by individual cattle MHC class II DR (BoLA-DR) molecules was studied in a direct MHC-peptide binding assay. By in vitro priming of T lymphocytes derived from animals homozygous for both MHC class I and II, five T-cell epitopes were analyzed in the context of three MHC class II haplotypes. We found that the presentation of these T-cell epitopes was mediated by DR molecules, since blocking this pathway of antigen presentation using monoclonal antibody TH14B completely abolished the proliferative responses against the peptides. To study the DR-restricted presentation of these T-cell epitopes, a direct MHC-peptide binding assay on isolated cattle DR molecules was developed. Purified cattle MHC class II DR molecules of the BoLA-DRB3*0201, BoLA-DRB3*1101, and BoLADRB3*1201 alleles were isolated from peripheral blood mononuclear cells. For each allele, one of the identified T-cell epitopes was biotinylated, and used as a marker peptide for the development of a competitive MHC-peptide binding assay. Subsequently, the T-cell epitopes of FMDV with functionally defined MHC class II specificity were analyzed in this binding assay. The affinity of the epitopes to bind to certain DR molecules was significantly correlated to the capacity to induce T-cell proliferation This demonstrated at the molecular level that the selection of individual T-cell epitopes found at the functional level was indeed the result of MHC restriction.