STRUCTURE OF HEPATITIS B ANTIGENS

目的是获得有关结构的详细资料， B型肝炎病毒蛋白，并确定它们之间的关系， 蛋白质结构及其抗原功能。 这些研究将包括 HBsAg的研究，包括前S基因产物、HBeAg、HBcAg、 X蛋白 HBsAg的研究将包括对完整HBsAg的研究。 颗粒及其蛋白质和脂质成分。 我们将利用我们的专家组 单克隆抗-HBs抗体（含有抗组特异性， 抗亚型特异性抗体）结合氨基酸序列 分析、化学修饰研究、合成肽、免疫印迹 技术，寡核苷酸定点，特异性诱变，和 真核表达载体，用于从克隆的 病毒基因，试图确定HBsAg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 显微镜低角x射线散射，傅立叶变换红外 分光光度法和圆二色性，以获得关于 蛋白质亚基的数量、排列和二级结构， HBsAg以及脂质成分如何与此维持相关 结构 前S蛋白在颗粒结构中的作用， 还将研究抗原活性。 将研究HBcAg转化为HBeAg的化学过程， 化学转化与抗原改变相关。 位置 将通过与所用相同的方法寻找相关抗原位点 对于HBsAg。 我们还将使用抗合成肽抗体， 试图描述病毒“X”基因的产物 哺乳动物 表达载体将用于合成蛋白质， 进一步表征。 人类对HBV感染或HBV感染后产生的抗体。 通过目前的疫苗（Heptavax）以及 对实验性酵母HBsAg疫苗的反应将通过以下方法进行检查： 竞争研究与我们的单克隆抗体，以确定 这些人抗体的相对量和特异性。 是 预计这将有助于确定主要的免疫原性决定， 被人类所认识。

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.