鸡脾脏白髓结构特征及其淋巴细胞归巢的初步研究

发布时间：2018-04-19 03:37

本文选题：鸡脾 + 白髓　；参考：《南京农业大学》2015年硕士论文

【摘要】：鸡作为我国传统养殖的家禽,已经有数千年历史。随着饲养的进步和社会需求增大,密集化养殖的优点日益突出,但与此同时引起鸡发病率上升的问题也不容忽视。脾脏作为鸡内体最大的外周免疫器官,相关的研究较少。本文在研究鸡脾脏白髓的结构特征的基础上,分离外周血淋巴细胞并用荧光素羧基荧光素二醋酸盐琥珀酰亚胺酯(5,6-carboxy fluoresceindiacetate succinimidyl ester,CFSE)对其进行标记。以示踪其在脾脏中归巢的过程,探讨鸡淋巴细胞在脾脏归巢的意义,为鸡病的防治提供一定的理论依据·具体研究内容和结果如下:1.鸡脾脏白髓的结构特征脾脏作为鸡的主要免疫器官之一,其内在显微结构特征的研究具有重要意义。通过观察鸡脾脏椭球及笔毛微动脉的结构特征,探讨其与免疫反应的相关联系。应用光镜技术观察、研究鸡脾脏椭球结构,发现椭球内的笔毛微动脉内皮为立方高柱状内皮,细胞连接不紧密,并有淋巴细胞出现在椭球壁中·推断椭球中的笔毛微动脉可能是鸡淋巴细胞在脾脏发生归巢的场所。2.鸡外周血淋巴细胞的分离及CFSE标记随着研究技术的改进,有关细胞体内示踪的方法增多,其中利用CFSE标记来进行细胞体内示踪试验的效果较好。本试验应用梯度离心与CFSE荧光标记技术,对外周血淋巴细胞进行荧光标记。发现利用人淋巴分离液可以有效的分离鸡外周血淋巴细胞,CFSE标记外周血淋巴细胞活力良好,形态正常,荧光显微镜下可见大部分被标记细胞发出绿色荧光,阳性标记率高达98%。说明梯度离心分离外周血淋巴细胞确实可行,使用荧光染料CFSE进行活细胞体内示踪是一种稳定的示踪方法。3.鸡脾脏淋巴细施归巢示踪的相关研究将被标记的淋巴细胞注入鸡体内后,发现其中部分位于脾脏红髓,部分位于椭球周围淋巴鞘(PELS)内部或者附近。另外,观察到少部分淋巴细胞位于白髓椭球中央。试验结果表明被荧光标记的淋巴细胞能进入脾脏白髓内,鸡脾脏确实存在淋巴细胞归巢现象,并且与鞘毛细血管外的椭球密切相关。
[Abstract]:Chicken as a traditional poultry in China, has a history of thousands of years.With the progress of raising and the increasing of social demand, the advantages of intensive farming are becoming more and more prominent, but at the same time, the problem caused by the rising incidence of chicken can not be ignored.Spleen is the largest peripheral immune organ in chicken.On the basis of studying the structural characteristics of chicken spleen white pulp, peripheral blood lymphocytes were isolated and labeled with fluorescein carboxyfluorescein diacetate succinimide 56-carboxy fluoresceindiacetate succinimidyl succinimidyl.In order to trace the homing process of chicken lymphocytes in spleen, the significance of chicken lymphocytes homing in spleen was discussed, which provided some theoretical basis for the prevention and treatment of chicken disease. The specific research contents and results were as follows: 1.The structural characteristics of White Pulp in Chicken spleen as one of the main immune organs of Chicken, the study of its intrinsic microstructure is of great significance.By observing the structural characteristics of chicken spleen ellipsoid and pencil-hairy arterioles, the relationship between spleen ellipsoid and immune response was discussed.The structure of chicken spleen ellipsoid was studied by light microscopy. It was found that the endothelium of the penomaricular artery in the ellipsoid was cubic high columnar endothelium, and the cells were not closely connected.Moreover, some lymphocytes appeared in the ellipsoid wall. It was inferred that the pencil-capillary artery in the ellipsoid might be the place where chicken lymphocytes homing in the spleen.Isolation of chicken peripheral blood lymphocytes and CFSE labeling with the improvement of research technology, the methods of cell in vivo tracer were increased, and the results of in vivo tracer test with CFSE labeling were better.In this experiment, gradient centrifugation and CFSE fluorescence labeling were used to label peripheral blood lymphocytes.It was found that using human lymphoid isolate could effectively isolate chicken peripheral blood lymphocytes labeled with CFSE with good viability and normal morphology. Under fluorescence microscope, most labeled cells could emit green fluorescence, and the positive labeling rate was as high as 98%.The results showed that the separation of peripheral blood lymphocytes by gradient centrifugation was feasible. Using fluorescent dye CFSE to trace living cells in vivo was a stable tracer method.Some of the labeled lymphocytes were injected into the chicken body and some of them were located in the red pulp of the spleen and partly in or near the peri-ellipsoid lymphatic sheath (PELSs).In addition, a small number of lymphocytes were observed at the center of the white pulp ellipsoid.The results showed that the fluorescent labeled lymphocytes could enter the white marrow of the spleen and the chicken spleen did have the phenomenon of lymphocyte homing which was closely related to the ellipsoid outside the sheath capillaries.
【学位授予单位】：南京农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S831

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