Carboxyl-terminal domain of the Epstein-Barr virus nuclear antigen is highly immunogenic in man.
Carboxyl-terminal domain of the Epstein-Barr virus nuclear antigen is highly immunogenic in man.
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Epstein-Barr 病毒核抗原的羧基末端结构域在人体中具有高度免疫原性。
DOI：
10.1073/pnas.82.18.6300
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发表时间：
1985
影响因子：
11.1
通讯作者：
Hayward,SD
中科院分区：
文献类型：
--
作者：
Milman,G;Scott,AL;Cho,MS;Hartman,SC;Ades,DK;Hayward,GS;Ki,PF;August,JT;Hayward,SD
利用高表达质粒在大肠杆菌中合成了BamHI限制性内切片段K编码的eb病毒核抗原（EBNA-1）的羧基末端1 / 3。所得的28 kda EBNA融合多肽，含可溶性细菌蛋白总量的5-10%，通过磷酸纤维素和羟基磷灰石柱层析纯化到明显的均匀性。兔抗28-kDa EBNA单克隆抗体和小鼠抗28-kDa EBNA单克隆抗体对eb病毒感染的淋巴母细胞样细胞系进行核免疫荧光染色，在免疫印迹上识别出相应的完整EBNA多肽带。以纯化的28-kDa EBNA为抗原，采用ELISA法定量测定人血清中EBNA抗体。ELISA法的灵敏度比经典的抗补体免疫荧光法高约100倍。在病毒衣壳抗原血清阳性的正常人血清中检测到抗ebna抗体，而在大多数急性感染性单核细胞增多症患者血清中检测到低抗ebna滴度。该试验给出了类风湿关节炎、伯基特淋巴瘤或鼻咽癌患者血清中预期的滴度模式，从而证实了该纯化试剂用于评估EBNA抗体状态的有效性。大约10%的正常人和类风湿关节炎患者的抗ebna滴度与鼻咽癌患者一样高。在这些高滴度的个体中，大于1%的IgG抗体是识别28-kDa EBNA的抗体，这表明EBNA的羧基末端结构域具有高度的免疫原性。
The carboxyl-terminal one-third of the Epstein-Barr virus nuclear antigen (EBNA-1) encoded by the BamHI restriction fragment K was synthesized in Escherichia coli by use of a high-expression plasmid. The resultant 28-kDa EBNA fusion polypeptide, comprising 5-10% of the total soluble bacterial protein, was purified to apparent homogeneity by phosphocellulose and hydroxylapatite column chromatography. Both rabbit monospecific antibodies and mouse monoclonal antibodies against 28-kDa EBNA gave nuclear immunofluorescence staining on Epstein-Barr virus (EBV)-infected lymphoblastoid cell lines and recognized the appropriate intact EBNA polypeptide bands on immunoblots. An ELISA with the purified 28-kDa EBNA as antigen was used to quantitate anti-EBNA antibody in human serum samples. The ELISA method was approximately 100-fold more sensitive than the classical anticomplement immunofluorescence assay. Anti-EBNA antibody was detected in sera from 100% of normal individuals who were seropositive for the viral capsid antigen, and low anti-EBNA titers were detected in serum from most patients with acute infectious mononucleosis. The assay gave the expected pattern of titers in sera from patients with rheumatoid arthritis, Burkitt lymphoma, or nasopharyngeal carcinoma, thus confirming the validity of this purified reagent for assessing EBNA antibody status. Approximately 10% of normal individuals and rheumatoid arthritis patients had anti-EBNA titers as high as those seen in nasopharyngeal carcinoma patients. In these high-titer individuals, greater than 1% of the total IgG are antibodies that recognize 28-kDa EBNA, which indicates that the carboxyl-terminal domain of EBNA is highly immunogenic.