Bungy Poor Fard G A, Latchman Y, Rodda S, Geysen M, Roitt I, Brostoff J
Department of Immunology, University College London Medical School, UK.
Clin Exp Immunol. 1993 Oct;94(1):111-6. doi: 10.1111/j.1365-2249.1993.tb05986.x.
One hundred and fifteen overlapping synthetic peptides spanning the entire sequence of the iso-allergen clone1A of Lol p I from rye grass Lolium perenne were synthesized by the multi-pin technique. The peptides were overlapping 12mers, offset by two residues and overlapping by 10 residues. Sets of six adjacent overlapping peptides (except pool-1, 15, 20) were pooled and were used in vitro and in vivo to map the T cell epitopes on Lol p I. Six atopics who were skin test and RAST positive to rye grass showed T cell responses to L. perenne extract (LPE) and its major fraction (Lol p I). Five out of six showed T cell responses in vitro to peptide pool-17, while five non-atopics did not respond to any of the peptide pools. By testing the individual peptides of pool-17, we have located the T cell epitope on Lol p I. Interestingly, when we tested pool-17 and its single peptides in vivo by intradermal skin testing we found in one patient a typical DTH after 24-48 h to pool-17 and its peptides (peptides 3 and 4) which exactly matched the in vitro responses. By defining the T cell epitopes in this way a greater understanding of the allergic response to pollen will be obtained, and a more effective and less dangerous vaccine may be possible for treating patients with hay fever.
采用多针技术合成了115个重叠的合成肽,这些肽覆盖了黑麦草(Lolium perenne)中Lol p I同种变应原克隆1A的整个序列。这些肽是重叠的12肽,偏移两个残基,重叠10个残基。将六组相邻的重叠肽(池1、15、20除外)合并,用于体外和体内定位Lol p I上的T细胞表位。6名对黑麦草皮肤试验和RAST呈阳性的特应性个体显示出对多年生黑麦草提取物(LPE)及其主要成分(Lol p I)的T细胞反应。6名个体中有5名在体外对肽池17有T细胞反应,而5名非特应性个体对任何肽池均无反应。通过测试池17中的单个肽,我们定位了Lol p I上的T细胞表位。有趣的是,当我们通过皮内皮肤试验在体内测试池17及其单个肽时,我们在一名患者中发现,在24 - 48小时后,对池17及其肽(肽3和4)出现了典型的迟发型超敏反应,这与体外反应完全匹配。通过这种方式定义T细胞表位,将能更深入地了解对花粉的过敏反应,并且有可能开发出一种更有效且危险性更低的疫苗来治疗花粉热患者。