Hong S S, Bardy M, Monteil M, Gay B, Denesvre C, Tournier J, Martin G, Eloit M, Boulanger P
Laboratoire de Virologie et Pathogénèse Virale, Faculté de Médecine, Lyon, France.
Viral Immunol. 2000;13(3):353-71. doi: 10.1089/08828240050144671.
A panel of nine independent mouse monoclonal antibodies (MAbs) against penton base capsomers of subgenus C adenovirus serotypes 2 (Ad2) and 5 (Ad5) were isolated and characterized. Two of them (1D2 and 5A5), raised against Ad5 virion as the immunogen, bound to sodium dodecyl sulfate (SDS)-resistant and subgenus C-specific epitopes that were not present in subgenus B Ad3 penton base. The 1D2 and 5A5 epitopes were mapped to two distinct regions that did not belong to the main variable region carrying the integrin-binding RGD motif at position 340. For the other seven MAbs, raised against recombinant Ad2 penton base protein (9S-pentamers), the epitopes were sensitive to SDS-denaturation, but reacted with native Ad2, Ad5, and Ad3 penton base. The epitopes recognized by the nine MAbs and by polyclonal antipenton base antibodies defined three major immunoreactive regions. One (I) mapped to the N-terminal domain (residues 116-165); the other two regions were almost symmetrically disposed on both sides of the integrin-binding RGD motif at position 340, within residues 248-270 (II), and within residues 368-427 (III) in the C-terminal domain. Region II overlapped the fiber-binding site in penton base (residues 254-260). None of the MAbs showed any detectable virus neutralization effect, but they all slightly augmented the efficiency of Ad-mediated gene transfer. Although none of their epitopes included the RGD-340 tripeptide, substitutions of the arginine residue in the RGD motif abolished the reactivity of six individual and distant epitopes, suggesting a major conformational role for the RGD-containing domain.
分离并鉴定了一组针对C亚群腺病毒血清型2（Ad2）和5（Ad5）五邻体基底壳粒的9种独立小鼠单克隆抗体（MAb）。其中两种（1D2和5A5）以Ad5病毒粒子作为免疫原产生，它们与十二烷基硫酸钠（SDS）抗性且C亚群特异性的表位结合，这些表位不存在于B亚群Ad3五邻体基底中。1D2和5A5表位被定位到两个不同区域，它们不属于在第340位携带整合素结合RGD基序的主要可变区。对于另外七种以重组Ad2五邻体基底蛋白（9S - 五聚体）作为免疫原产生的MAb，其表位对SDS变性敏感，但能与天然的Ad2、Ad5和Ad3五邻体基底发生反应。这9种MAb和多克隆抗五邻体基底抗体识别的表位定义了三个主要免疫反应区域。一个区域（I）定位到N端结构域（第116 - 165位氨基酸残基）；另外两个区域几乎对称地分布在第340位整合素结合RGD基序两侧，分别位于C端结构域的第248 - 270位氨基酸残基（II）和第368 - 427位氨基酸残基（III）。区域II与五邻体基底中的纤维结合位点（第254 - 260位氨基酸残基）重叠。这些MAb均未显示出任何可检测到的病毒中和作用，但它们都略微提高了Ad介导的基因转移效率。尽管它们的表位均不包含RGD - 340三肽，但RGD基序中精氨酸残基的替换消除了六个单独且距离较远的表位的反应性，表明含RGD结构域具有主要的构象作用。
