Noraz N, Benichou S, Madaule P, Tiollais P, Vernant J C, Desgranges C
Inserm U271, Lyon, France.
Virology. 1993 Mar;193(1):80-8. doi: 10.1006/viro.1993.1105.
A peptide library of HTLV-I Env and Tax proteins was constructed in yeast in order to characterize which domains of these proteins are immunogenic in HTLV-I-infected individuals. Five yeast colonies (A to E) were selected using HTLV-I positive plasma, and one yeast colony (F) was selected using rabbit anti-Tax sera. Plasmid DNA present in each positive clone was recovered and sequenced. Overlapping clones A to E covered the C-terminal part of the gp46 exterior glycoprotein (aa 197 to 305) and were all glycosylated. Clone F encoded the C-terminal 25 aa of Tax (aa 329-353). Recombinant peptides were recognized by more than 40% of the HTLV-I positive human sera, confirming that they are major immunodominant domains. We studied the antibody response to each recombinant peptide in patients with TSP/HAM and asymptomatic carriers. Higher absorbance values were obtained with sera from TSP/HAM patients than from asymptomatic carriers, but the difference between the two groups was not statistically significant. Our study confirms that the COOH-terminal region of gp46 is highly immunogenic in HTLV-I-infected individuals and demonstrates a new immunogenic epitope of the Tax protein.
构建了人嗜T淋巴细胞病毒I型(HTLV-I)包膜蛋白(Env)和反式激活蛋白(Tax)的肽库,以确定这些蛋白的哪些结构域在HTLV-I感染个体中具有免疫原性。使用HTLV-I阳性血浆筛选出5个酵母菌落(A至E),并使用兔抗Tax血清筛选出1个酵母菌落(F)。回收每个阳性克隆中的质粒DNA并进行测序。重叠克隆A至E覆盖了gp46外膜糖蛋白的C末端部分(第197至305位氨基酸),且均被糖基化。克隆F编码Tax的C末端25个氨基酸(第329 - 353位氨基酸)。超过40%的HTLV-I阳性人血清可识别重组肽,证实它们是主要的免疫显性结构域。我们研究了热带痉挛性截瘫/HTLV-I相关脊髓病(TSP/HAM)患者和无症状携带者对每种重组肽的抗体反应。TSP/HAM患者血清的吸光度值高于无症状携带者,但两组之间的差异无统计学意义。我们的研究证实,gp46的COOH末端区域在HTLV-I感染个体中具有高度免疫原性,并展示了Tax蛋白的一个新的免疫原性表位。