本文選題：雞脾 + 白髓��； 參考：《南京農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：雞作為我國傳統(tǒng)養(yǎng)殖的家禽,已經(jīng)有數(shù)千年歷史。隨著飼養(yǎng)的進步和社會需求增大,密集化養(yǎng)殖的優(yōu)點日益突出,但與此同時引起雞發(fā)病率上升的問題也不容忽視。脾臟作為雞內(nèi)體最大的外周免疫器官,相關(guān)的研究較少。本文在研究雞脾臟白髓的結(jié)構(gòu)特征的基礎(chǔ)上,分離外周血淋巴細胞并用熒光素羧基熒光素二醋酸鹽琥珀酰亞胺酯(5,6-carboxy fluoresceindiacetate succinimidyl ester,CFSE)對其進行標記。以示蹤其在脾臟中歸巢的過程,探討雞淋巴細胞在脾臟歸巢的意義,為雞病的防治提供一定的理論依據(jù)·具體研究內(nèi)容和結(jié)果如下:1.雞脾臟白髓的結(jié)構(gòu)特征脾臟作為雞的主要免疫器官之一,其內(nèi)在顯微結(jié)構(gòu)特征的研究具有重要意義。通過觀察雞脾臟橢球及筆毛微動脈的結(jié)構(gòu)特征,探討其與免疫反應(yīng)的相關(guān)聯(lián)系。應(yīng)用光鏡技術(shù)觀察、研究雞脾臟橢球結(jié)構(gòu),發(fā)現(xiàn)橢球內(nèi)的筆毛微動脈內(nèi)皮為立方高柱狀內(nèi)皮,細胞連接不緊密,并有淋巴細胞出現(xiàn)在橢球壁中·推斷橢球中的筆毛微動脈可能是雞淋巴細胞在脾臟發(fā)生歸巢的場所。2.雞外周血淋巴細胞的分離及CFSE標記隨著研究技術(shù)的改進,有關(guān)細胞體內(nèi)示蹤的方法增多,其中利用CFSE標記來進行細胞體內(nèi)示蹤試驗的效果較好。本試驗應(yīng)用梯度離心與CFSE熒光標記技術(shù),對外周血淋巴細胞進行熒光標記。發(fā)現(xiàn)利用人淋巴分離液可以有效的分離雞外周血淋巴細胞,CFSE標記外周血淋巴細胞活力良好,形態(tài)正常,熒光顯微鏡下可見大部分被標記細胞發(fā)出綠色熒光,陽性標記率高達98%。說明梯度離心分離外周血淋巴細胞確實可行,使用熒光染料CFSE進行活細胞體內(nèi)示蹤是一種穩(wěn)定的示蹤方法。3.雞脾臟淋巴細施歸巢示蹤的相關(guān)研究將被標記的淋巴細胞注入雞體內(nèi)后,發(fā)現(xiàn)其中部分位于脾臟紅髓,部分位于橢球周圍淋巴鞘(PELS)內(nèi)部或者附近。另外,觀察到少部分淋巴細胞位于白髓橢球中央。試驗結(jié)果表明被熒光標記的淋巴細胞能進入脾臟白髓內(nèi),雞脾臟確實存在淋巴細胞歸巢現(xiàn)象,并且與鞘毛細血管外的橢球密切相關(guān)。
[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.
【學(xué)位授予單位】：南京農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S831

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