OPERATIONAL AND TOPOLOGICAL ANALYSES OF ANTIGENIC SITES ON INFLUENZA-C VIRUS GLYCOPROTEIN AND THEIR DEPENDENCE ON GLYCOSYLATION
OPERATIONAL AND TOPOLOGICAL ANALYSES OF ANTIGENIC SITES ON INFLUENZA-C VIRUS GLYCOPROTEIN AND THEIR DEPENDENCE ON GLYCOSYLATION
复制标题
DOI：
10.1099/0022-1317-69-3-537
复制
发表时间：
1988-03-01
影响因子：
3.8
通讯作者：
NAKAMURA, K
中科院分区：
文献类型：
--
作者：
SUGAWARA, K;KITAME, F;NAKAMURA, K
本实验室制备了7株抗丙型流感病毒糖蛋白（gp 88）的单克隆抗体，初步分为两组：A组（J14，J 9，Q5，K16）具有中和活性，而B组（S16，J6，J15）无中和活性。这些抗体用于分析gp 88的抗原结构，并检查糖基化对糖蛋白抗原性的影响。用A组抗体中的每一种选择的一组抗原变体的操作分析鉴定了gp 88分子上的两个不重叠的抗原位点，位点A-1被J14、J 9和Q5识别，位点A-2被K16识别。位点A-1和A-2显示，但是，通过竞争性结合试验，地形重叠。与B组抗体的竞争性结合分析鉴定了两个额外的非重叠抗原位点，S16识别的位点B-1和J6和J15识别的位点B-2。在放射免疫沉淀实验中发现，位点B-1和B-2的抗体不仅与gp 88反应，而且与在衣霉素存在下合成的其非糖基化形式（T76）反应。相反，位点A-1和A-2的抗体仅以痕量或根本不免疫沉淀T76多肽。此外，Western印迹分析表明，在硝酸纤维素上印迹的变性gp 88与位点B-1和B-2的抗体反应，但不与位点A-1和A-2的抗体反应。这些观察结果表明，糖基化的gp 88选择性地影响抗原位点A-1和A-2的完整性，这是由构象依赖性表位。
In our previous study, seven monoclonal antibodies specific for influenza C virus glycoprotein (gp88) were prepared and tentatively classified into two groups: group A (J14, J9, Q5, K16) has neutralization activity whereas group B (S16, J6, J15) does not. These antibodies were used to analyse the antigenic structure of gp88 and to examine the effect of glycosylation on the antigenicity of the glycoprotein. Operational analysis with a panel of antigenic variants selected with each of the group A antibodies identified two non-overlapping antigenic sites on the gp88 molecules, site A-1 recognized by J14, J9 and Q5 and site A-2 by K16. Sites A-1 and A-2 were shown, however, to be topographically overlapping by competitive binding assays. Competitive binding analysis with group B antibodies identified two additional non-overlapping antigenic sites, site B-1 recognized by S16 and site B-2 by J6 and J15. It was found in radioimmunoprecipitation experiments that antibodies to sites B-1 and B-2 were reactive not only with gp88 but with its non-glycosylated form (T76) synthesized in the presence of tunicamycin. Antibodies to sites A-1 and A-2, in contrast, immunoprecipitated the T76 polypeptide in only trace amounts or not at all. Additionally, Western blot analysis showed that denatured gp88 blotted on nitrocellulose was reactive with antibodies to sites B-1 and B-2 but not with those to sites A-1 and A-2. These observations suggest that glycosylation of gp88 selectively influences the integrity of antigenic sites A-1 and A-2 which are composed of conformation-dependent epitopes.