Jenison S A, Yu X P, Valentine J M, Galloway D A
Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
J Virol. 1989 Feb;63(2):809-18. doi: 10.1128/JVI.63.2.809-818.1989.
Recombinant proteins encoded by the human papillomavirus type 6b (HPV6b) L1 open reading frame react with sera from patients with condylomata acuminata and also react with rabbit antiserum raised against sodium dodecyl sulfate-disrupted bovine papillomavirus type 1 (BPV1) virions. To map the immunoreactive epitopes, a series of procaryotic expression plasmids was made which contained a nested set of 3' to 5' deletions in the HPV6b L1 open reading frame. The deleted plasmids expressed a set of carboxy to amino terminus truncated fusion proteins. Regions containing the immunoreactive epitopes were mapped by determining which of the deleted fusion proteins retained reactivity with sera in Western immunoblot assays. The coding sequence for a human antibody-reactive linear epitope mapped between HPV6b nucleotide coordinates 7045 and 7087, and the rabbit anti-BPV1-reactive epitope coding sequence mapped between coordinates 6377 and 6454. Synthetic peptides derived from the epitope mapping were reacted with sera in enzyme-linked immunosorbent assay. Human sera reacted with synthetic peptide QSQAITCQKPTPEKEKPDPYK (HPV6b L1 amino acids 417 through 437). Rabbit anti-BPV1 and rabbit antisera raised against HPV16 L1 recombinant proteins reacted with the synthetic peptide DGDMVDTGFGAMNFADLQTNKSDVPIDI (HPV6b L1 amino acids 193 through 220). Human sera which reacted with HPV6b L1 fusion proteins cross-reacted with an HPV11 L1 fusion protein but did not react with fusion proteins encoded by HPV1a, HPV16, or HPV18. Rabbit anti-BPV1 reacted with L1 fusion proteins encoded by all of these HPV types. In contrast to the type-common (rabbit anti-BPV1-reactive) epitope, the human antibody-reactive epitope appears to be relatively HPV type specific.
人乳头瘤病毒6b型（HPV6b）L1开放阅读框编码的重组蛋白可与尖锐湿患者血清发生反应，也可与针对十二烷基硫酸钠破坏的1型牛乳头瘤病毒（BPV1）病毒粒子产生的兔抗血清发生反应。为了定位免疫反应性表位，构建了一系列原核表达质粒，这些质粒在HPV6b L1开放阅读框中包含一组从3'到5'的嵌套缺失。缺失的质粒表达了一组从羧基端到氨基端截短的融合蛋白。通过确定哪些缺失的融合蛋白在Western免疫印迹分析中与血清保持反应性，来定位包含免疫反应性表位的区域。人抗体反应性线性表位的编码序列定位于HPV6b核苷酸坐标7045和7087之间，兔抗BPV1反应性表位编码序列定位于坐标6377和6454之间。从表位定位衍生的合成肽在酶联免疫吸附测定中与血清反应。人血清与合成肽QSQAITCQKPTPEKEKPDPYK（HPV6b L1氨基酸417至437）发生反应。兔抗BPV1和针对HPV16 L1重组蛋白产生的兔抗血清与合成肽DGDMVDTGFGAMNFADLQTNKSDVPIDI（HPV6b L1氨基酸193至220）发生反应。与HPV6b L1融合蛋白发生反应的人血清与HPV11 L1融合蛋白发生交叉反应，但不与HPV1a、HPV16或HPV18编码的融合蛋白发生反应。兔抗BPV1与所有这些HPV类型编码的L1融合蛋白发生反应。与类型共同（兔抗BPV1反应性）表位不同，人抗体反应性表位似乎相对具有HPV类型特异性。
