树鼩NKT细胞新型抗原递呈分子CD1d的分子克隆及初步分析
文献类型：学位论文
| 作者 | 张萍 |
| 学位类别 | 硕士 |
| 答辩日期 | 2008-06 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 北京 |
| 导师 | 徐小宁 |
| 关键词 | 树鼩 HBV NKT细胞 CD1d 系统进化树 组织表达 |
| 其他题名 | Molecular cloning and sequence analyse of CD1d from tree shrews |
| 学位专业 | 细胞生物学 |
| 中文摘要 | 目前多数学者认为树鼩（Tree shrews）是灵长目攀居目树鼩科的一种小型哺乳动物。因其具有体型小、易饲养、价格低、新陈代谢和解剖结构与人类非常接近等优点，树鼩已经成为多种人类疾病研究的良好动物模型。树鼩作为乙型肝炎病毒（HBV）感染的动物模型已经被广泛应用。 自然杀伤性T细胞（Natrual killer T cells, NKT cells）可同时表达NK细胞和T细胞的表面标记，是CD1d限制性的特殊的淋巴细胞亚群，在机体的免疫反应中起重要作用。研究表明，在HBV感染及乙型肝炎的发病过程中，NKT细胞既有抑制和清除病毒的作用，也有加重肝炎及相关病症的负作用。因此，对NKT细胞的研究及恰当地调节将有助于理解和解决在研究乙肝防治中遇到的问题，但在树鼩中，至今尚无NKT细胞的相关报道。 目前公认的检测NKT细胞的手段是用加载了α-GalCer的CD1d四聚体检测。但至今尚无树鼩特异的相关抗体，给树鼩NKT细胞的研究造成很大障碍。因此，获得树鼩特异的CD1d四聚体是研究其NKT细胞特征和功能的前提。本论文作为其中一部分，目的是克隆树鼩CD1d编码区全长序列，为今后抗体的制备奠定基础。 本论文中，我们完成了相关工作并得到了以下结果：1、首次克隆得到了树鼩CD1d基因编码区的全长序列（1002bp），并预测了其氨基酸序列；2、通过比对分析得知其与灵长类的CD1d同源性较高，达到80％以上；同时，通过系统进化树分析，得知树鼩与灵长类亲缘关系较其它物种更近；3、首次分析了树鼩的CD1d mRNA在不同组织中的表达情况，发现树鼩的肝，肺中CD1d表达量较高，外周血和脾脏中也有少量表达。 总之，我们首次克隆得到了树鼩CD1d的编码区的全长序列，为今后制备CD1d四聚体进而研究树鼩的NKT细胞奠定了基础；同时首次分析了CD1d mRNA在树鼩的部分组织中的表达情况，结合以往其它物种中CD1d组织分布情况的检测结果，为树鼩中CD1d分子的研究提供了材料和数据基础。 |
| 英文摘要 | There exists a great controversy as to whether tree shrews should be placed in an order with primates or insectivores. However, tree shrews are considered as a closer relative of primates than rodents these years. With several advantages that they are smaller, easier to raise and that they are more similar to human in the metabolism and anatomic structure than other species, tree shrews have been widely used as an animal model in biomedical researches, such as infection, tumor, diabetes. We are establishing the chronic HBV infection model with tree shrew in our lab. NKT cells are a population of T cells that share some characteristics with NK cells and can be activated by the glycolipid antigens presented by CD1d, a nonclassical antigen-presenting molecule. Upon activation, NKT cells rapidly and abundantly produce both Th1 and Th2 cytokines, particularly IFN-γ and IL-4, which play important roles in innate and acquired immunity. It has been reported that therapeutic activation of NKT cells may represent a new strategy for the treatment of chronic HBV infection, while on the other hand, immune responses by intrahepatic NKT cells may enhance the liver injury to some extent. However, researche on NKT cells will be useful to understanding HBV infection and hepatitis controling in tree shrews. With difficulties that there are no specific CD1d-tetramer for NKT cells detecting in tree shrew, there’s no research on tree shrew NKT cells so far. We must firstly obtain the sequences of CD1d and β-microgloblin (β2-m) of tree shrews. The full length sequence β2-m has been reported, while CD1d not up to now, which is our main task in this thesis. The main work for this thesis is to obtain and analyse the full length sequence of CD1d coding region in tree shrews, including: 1. Cloning the correct sequence of tree shrew CD1d by PCR, sequencing and sequence assembly; 2. Identity analysis of CD1d from tree shrews and other species. Also, a phylogenetic tree was made based on CD1d coding region sequences to analyse the evolutionary relationship among different species and analyse the closest relative of tree shrews. 3. Analysis of tissue distribution of CD1d in tree shrews, rhesus monkeys and mice. The main research results were as follows: 1. We have obtained the full length sequence of tree shrew CD1d coding region which is 1002bp, coding 333 amino acids with a stop codon TGA. 2. The identity of CD1d between tree shrews and primates is up to 80%, higher than other species. From the phylogenetic tree based on CD1d sequences, we can see that tree shrew has a closest relationship with primates than with other species. 3. CD1d can be detected in liver, lung higher than in spleen and peripheral blood in tree shrews and different tissue distribution was detected in rhesus monkeys and mice. In a word, we have cloned the coding region of tree shrew CD1d and predicted its amino acid. Also, tissue distribution of CD1d was detected in tree shrews for the first time. These results offers possibility for the study on tree shrew NKT cells in future. |
| 语种 | 中文 |
| 公开日期 | 2010-10-13 |
| 源URL | [http://159.226.149.42:8088/handle/152453/6083]  |
| 专题 | 昆明动物研究所_其他 |
| 推荐引用方式 GB/T 7714 | 张萍. 树鼩NKT细胞新型抗原递呈分子CD1d的分子克隆及初步分析[D]. 北京. 中国科学院研究生院. 2008. |
入库方式： OAI收割
来源：昆明动物研究所
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