骆驼肺胸膜下淋巴结结构与功能的研究

The mucosal-associated lymphoid tissue (MALT) of the lung in other animals includes diffuse lymphoid tissue and isolated lymphoid nodules，which are generally distributed along the bronchial. However, we observed that MALT of the bactrian camel is concentrated below the lung pleura, and consists of the lymph nodes. This unique structure never reported in other species. Moreover, its characteristic is not clear. To identify the special organs about their morphological and functional characteristics, we applied in this study via molecular biology techniques, immunological techniques, immunohistochemical techniques and electron microscopy techniques, etc. First, the immune morphological characteristics of the macroscopic, microscopic and submicroscopic level of the organs are clarified, and then the differences between the organs and the lymph nodes located in other parts of the bactrian camel are compared. To determine the relationship between the lymph nodes in the lung pleura and mucosal immunity, the monoclonal antibodies of SIgA extracted from the bactrian amel colostrum is prepared and the distribution of SIgA in the lung is detected. Additionally, the IgG, IgA, IgM, and IgE (humoral immune effector molecules )in the bactrian camel are cloned and expressed and their monoclonal antibody are also prepared to classify and locate the humoral immune effector cells , which further make us understand the relationship between the structure and humoral immune. The results will reveal both the immune morphological properties and functional properties of the special structure.

动物肺脏中的黏膜相关淋巴组织（MALT）通常是沿着支气管分布，以散在淋巴组织和淋巴小结的形式存在。但申请人在前期工作中发现，双峰驼的MALT与其他动物不同，集中分布于肺脏胸膜下，并以多个散在的淋巴结形式存在。其详细结构与功能尚不明了。因此，本项目拟采用分子生物学、免疫学和免疫组化技术以及电子显微镜等多种研究手段在宏观、微观及亚微观水平观测其构筑学特点，明确它与其他部位淋巴结，以及与黄牛、牦牛和羊肺MALT的差异。同时，从双峰驼初乳中提取黏膜免疫效应分子SIgA制备单抗，检测SIgA在肺脏中的分布，以此判断肺胸膜下淋巴结与黏膜免疫之间的关系；克隆与表达双峰驼体液免疫效应分子IgG、IgA、IgM和IgE重链特定区域基因，制备单抗，通过免疫组化技术对不同类型体液免疫效应细胞进行分类与定位，以了解该结构与体液免疫之间的关系。结果将阐明该结构的免疫形态学属性和功能属性，为黏膜免疫理论提供新的依据。

动物肺脏中的黏膜相关淋巴组织（MALT）通常是沿着支气管分布，以散在淋巴组织和淋巴小结的形式存在。但课题组在多年对动物黏膜免疫的研究中发现，双峰驼肺脏的MALT比较独特，有淋巴结存在。因此于2012年提出立项申请并获得资助，通过项目组全体成员的共同努力在多方面取得了原创性成果。项目研究首先通过分子生物学、免疫学、免疫组织化学以及电子显微镜等多种手段在宏观、微观及亚微观水平观测了双峰驼肺脏MALT的构筑学特点，同时从双峰驼初乳和血清中分别提取黏膜免疫重要效应分子SIgA和IgGs制备抗体，之后检测了2种抗体分泌细胞在呼吸系统和消化系统的分布特点，并进行统计分析。结果表明，双峰驼呼吸系统中的MALT主要集中在上呼吸道，在肺脏中还是以弥散淋巴组织为主，其中可见的淋巴结与年龄和个体等诸多因素密切相关，结构和其他部位淋巴结类似。SIgA和IgGs 2种抗体分泌细胞在整个呼吸道和消化道均有分布，但主要以SIgA抗体分泌细胞占优势。此外，该项目还获得了双峰驼IgGs 3个亚型的抗体。上述成果，对于动物黏膜免疫的深入研究与应用有着十分重要的理论价值和实际意义，也为后续的研究工作奠定了良好的工作基础。