STRUCTURE OF HEPATITIS B ANTIGENS

目的是获得有关结构的详细信息 乙肝病毒蛋白，并确定它们之间的关系 蛋白质结构及其抗原性功能。这些研究将包括 乙肝表面抗原的研究，包括S前基因产物、HBeAg、HBcAg和 “X”蛋白。对乙肝表面抗原的研究将包括对完整的 颗粒及其蛋白质和脂肪成分。我们将利用我们的小组 单抗-HBs抗体(包含抗群特异性和 抗亚型特异性抗体)与氨基酸序列结合 分析、化学修饰研究、合成肽、免疫印迹 技术，寡核苷酸定向位点，特异突变，和 克隆的乙型肝炎病毒表面抗原的真核表达载体 病毒基因试图确定乙肝表面抗原的物理结构和 各种抗原决定簇的位置和结构。我们还将 使用物理技术，如冷冻蚀刻和冷冻断裂电子 显微小角x射线散射、傅里叶变换红外 分光光度和圆二向色性，以获得关于 蛋白质亚基的数量、排列和二级结构 乙肝表面抗原及其脂类成分与其维持的关系 结构。前S蛋白在颗粒结构和细胞周期中的作用 还将调查其抗原性。 将研究HBcAg转化为HBeAg的化学，这 化学转化与抗原性改变相关。地点 将以相同的方法寻找相关的抗原点 对于乙肝表面抗原。我们还将在一种 试图描述病毒“X”基因的产物。哺乳动物 表达载体将被用来合成蛋白质以用于 进一步的刻画。 人类对乙肝病毒感染或对病毒的反应而产生的抗体 目前的疫苗(Heptavax)和#年生产的疫苗免疫 对实验性酵母乙肝疫苗的反应将通过以下方式进行检查 与我们的单抗进行竞争研究，以确定 这些人类抗体的相对数量和特异性。它是 预计这将有助于确定主要的免疫原性确定 为人类所认可。

The objective is to obtain detailed information concerning the structure of hepatitis B viral proteins, and to determine the relationship between their protein structure and their antigenic function. These studies will include research on the HBsAg, including the pre-S gene products, HBeAg, HBcAg and the "X" protein. Studies of the HBsAg will include studies of the intact particle and its protein and lipid components. We will utilize our panel of monoclonal anti-HBs antibodies (which contain anti-group specific and anti-subtypes specific antibodies) in conjunction with amino acid sequence analysis, chemical modification studies, synthetic peptides, immunoblotting techniques, olignucleotide directed site, specific mutagenesis, and eukaryotic expression vectors for the production of HBsAg from the cloned viral gene to attempt to determine the physical structure of HBsAg and the location and structure of the various antigenic determinants. We will also use physical techniques such as freeze ETCH and freeze fracture electron microscopy low angle x-ray scattering, fourier transform infrared spectorphotometry, and circular dichroism to gain information on the number, arrangement, and secondary structure of the protein subunits with HBsAg and how the lipid components are related to the maintenance of this structure. The role of the pre-S proteins in the particle structure and antigenic activity will also be investigated. The chemistry of the conversion of HBcAg to HBeAg will be studied and this chemical conversion correlated with the antigenic alteration. The location of the relevant antigenic sites will be sought by the same methods as used for HBsAg. We will also use anti-synthetic peptide antibodies in an attempt to characterize the product of the viral "X" gene. Mammalian expression vectors will be used in an effort to synthesize the protein for further characterization. The antibody produced in humans in response to HBV infection or to immunization by the current vaccine (Heptavax) as well as that produced in response to the experimental yeast HBsAg vaccines will be examined by competition studies with our monoclonal antibodies to determine the relative amounts and specificities of these human antibodies. It is expected that this will help define the major immunogenic determination recognized by humans.