Fine mapping and characterisation of polymorphic immunoregulatory genes in the central MHC

主要组织相容性复合体（MHC）是一组通常作为一个块遗传的基因。该组包括HLA基因，可以作为邻近基因的标记，这些基因的特征不太清楚。例如，HLA基因的一些变体形式（多态性）标志着对糖尿病、狼疮和伊加缺乏症的易感性的差异。我们认为，这可能是由邻近的未表征基因的变化引起的，这些基因影响了对各种靶器官持续性炎症的控制。到目前为止的证据表明，中央MHC中的一个基因可能是负责的。在这个项目中，我们将研究来自具有HLA和其他MHC基因的不寻常组合的患者的DNA，以进一步确定MHC的哪一部分包含关键的免疫调节基因。MHC中的大多数基因现在已经被人类基因组计划识别，因此我们将能够在感兴趣的区域中选择最有希望的候选者。然后，我们将使用已知HLA类型的DNA来确定候选基因是否在个体之间存在差异。然后通过创建以反向（反义）方向携带部分基因的细胞系来研究感兴趣的基因的功能。这产生了一种反向信使（m）RNA，它结合正常的mRNA并阻止蛋白质的合成。然后，我们将检查所得细胞系的哪些反应是异常的（例如：炎症介质或细胞因子的产生）。在阐明了我们基因的功能之后，我们将在培养的细胞中过度表达患者中发生的每个版本，并寻找功能上的差异。在这项工作的同时，我们将使用具有已知MHC基因组合的实验室小鼠来建立特定基因在活体动物中的作用。

The major histocompatibility complex (MHC) is a group of genes that is usually inherited as a block. The group includes the HLA genes which can serve as markers for neighbouring genes that are less well characterised. For example, some variant forms (polymorphisms) of the HLA genes mark differences in susceptibility to diabetes, lupus and IgA deficiency. We propose that this may be caused by variations in the neighbouring uncharacterised genes affecting control of persistent inflammation of various target organs. Evidence so far suggests a gene in the central MHC may be responsible. In this project we will study DNA from patients who have unusual combinations of HLA and other MHC genes, to further define which part of the MHC contains the critical immunoregulatory genes. Most genes in the MHC have now been identified by the Human Genome Project, so we will be able to select the most promising candidates in the region of interest. We will then use DNA of known HLA types to determine if the candidate genes vary between individuals. The function of interesting genes will then be investigated by creating cell lines carrying part of the gene in the reverse (anti-sense) orientation. This generates a reverse messenger (m) RNA which binds the normal mRNA and prevents synthesis of the protein. We will then examine which responses of the resultant cell lines are abnormal (eg: production of inflammatory mediators or cytokines). Having elucidated the functions our genes, we will overexpress each version that occurs in patients in cultured cells and look for differences in function. In parallel with this work, we will use laboratory mice with known combinations of MHC genes to establish the effects of particular genes in a live animal.