Mäkelä M J, Lund G A, Salmi A A
Department of Medical Microbiology and Infectious Diseases, University of Alberta, Edmonton, Canada.
J Gen Virol. 1989 Mar;70 ( Pt 3):603-14. doi: 10.1099/0022-1317-70-3-603.
A combined analysis of hydrophilicity, accessibility and flexibility parameters of the deduced amino acid sequence of measles virus (MV) haemagglutinin (H) was used to select 10 regions for synthesis of 10- or 11-amino acid-long peptides. Nine of these sites are probably exposed on the surface of the protein, as polyclonal sera against either purified MV or purified H bound to these peptides as tested by enzyme immunoassay (EIA). Nevertheless, human sera from acute or chronic MV infection did not bind significantly to any peptide, indicating that the selected sites do not function as natural complete epitopes. All antisera raised in rabbits against keyhole limpet haemocyanin-conjugated peptides had a high titre to the homologous peptide and nine of them bound to MV lysate antigen, purified MV and/or purified H as tested in EIA. None of the sera had haemagglutination-inhibiting antibodies and only one antiserum (against peptide 185-195) had a neutralizing antibody titre of 1/160. Only a minority of the antisera were positive in Western blot (four of 10), radioimmunoprecipitation (two of 10) or immunofluorescence (three of 10). The results indicate that the computer program used in this analysis can predict surface-exposed areas of MV H but that the small peptides synthesized have little resemblance to natural antigenic sites.
通过对麻疹病毒（MV）血凝素（H）推导的氨基酸序列的亲水性、可及性和柔韧性参数进行综合分析，选择了10个区域来合成10或11个氨基酸长的肽段。通过酶免疫测定（EIA）测试，发现其中9个位点可能暴露于该蛋白表面，因为针对纯化的MV或纯化的H的多克隆血清与这些肽段结合。然而，来自急性或慢性MV感染患者的血清与任何肽段均无明显结合，这表明所选位点并非天然完整表位。所有用钥孔戚血蓝蛋白偶联肽段免疫家兔产生的抗血清对同源肽段均有高滴度，其中9种抗血清在EIA测试中与MV裂解物抗原、纯化的MV和/或纯化的H结合。所有血清均无血凝抑制抗体，只有一种抗血清（针对肽段185 - 195）的中和抗体滴度为1/160。只有少数抗血清在蛋白质印迹法（10种中有4种）、放射免疫沉淀法（10种中有2种）或免疫荧光法（10种中有3种）中呈阳性。结果表明，本分析中使用的计算机程序可以预测MV H的表面暴露区域，但合成的小肽与天然抗原位点几乎没有相似性。
