CELLULAR & MOLECULAR STUDIES OF LMP ANTIGENS & GENES

鼠LMP抗原是一组独特的蛋白质抗原 分子量范围为15- 30，000千道尔顿， 由主要组织相容性复合体（MHC）控制。 已经鉴定了16种不同的LMP抗原多肽 通过二维凝胶电泳， 显示了电泳多态性;这两种多态性 在H-2K和I-A基因座之间的MHC上。 所有16个 LMP链以大的、稳定的 胞内蛋白复合物 就像前面描述的三个 在MHC抗原的类别中，LMP抗原的表达是 由免疫干扰素（IFN-γ）调节，然而，LMP 抗原在生物化学、血清学和遗传学上是不同的 来自MHC I、II和III类抗原。 LMP复合体的功能及其与其他已知的 MHC抗原目前尚不清楚。 本文所述项目 利用两种独立的方法来收集线索， 这类新的MHC抗原的功能。 首先，兔抗血清 制备的纯化鼠LMP抗原将用于 免疫组织化学研究，以确定精确的亚细胞 LMP综合体的位置。 第二，克隆的DNA对应 LMP抗原亚基将从一组先前的 分离的重叠粘粒克隆跨越适当的 基因区域 这些基因组克隆将用于分离 cDNA克隆用于确定的一级结构， LMP抗原及其与已知的其他蛋白质的比较 功能，特别是紧密相连的MHC I类和II类抗原， 肿瘤坏死因子和光敏素。 希望了解一级结构和 LMP复合物的细胞内定位将提供其 生物功能，并允许设计特定的 生物测定以测试这些可能的功能。

The murine LMP antigens are a unique set of protein antigens ranging in molecular weight from 15-30,000 kilodaltons which are controlled by the major histocompatibility complex (MHC). Sixteen different LMP antigen polypeptides have been identified by two-dimensional gel electrophoresis, and two of these chains shown electrophoretic polymorphism; both of these polymorphisms map to the MHC, between the H-2K and I-A loci. All 16 of the LMP chains are associated with one another in a large, stable intracellular protein complex. Like the three previously described classes of MHC antigens, the expression of the LMP antigens is regulated by immune interferon (IFN-gamma), however, the LMP antigens are biochemically, serologically, and genetically distinct from MHC class I, II, and III antigens. The function of LMP complex and its relationship to other known MHC antigens is currently unknown. The project described herein utilizes two independent approaches for gathering clues to the function of this new class of MHC antigen. First, rabbit antisera made to purified murine LMP antigens will be used in immunohistochemical studies to determine the precise subcellular location of the LMP complex. Second, cloned DNA corresponding to LMP antigen subunits will be isolated from a set of previously isolated overlapping cosmid clones spanning the appropriate genetic region. These genomic clones will be used to isolate cDNA clones for a determination of the primary structure of the LMP antigens, and its comparison to other proteins of known function, especially the closely linked MHC class I and II antigens, tumor necrosis factor, and lymphotoxin. It is expected that knowledge of the primary structure and intracellular location of the LMP complex will yield clues to its biological function, and will permit the design of specific biological assays to test such possible functions.