【摘要】：本實(shí)驗(yàn)分別選擇18、20胚齡雞胚,1、4、7、10、14、21、35及56日齡的雛雞,每個(gè)時(shí)期取10只雞的新鮮回腸,制成冰凍切片,分別進(jìn)行HE染色和免疫組織化學(xué)染色。結(jié)合圖像分析和數(shù)據(jù)統(tǒng)計(jì),對(duì)雞回腸的組織結(jié)構(gòu)發(fā)育變化、淋巴細(xì)胞的出現(xiàn)、分布及定位規(guī)律做出研究。旨在了解不同時(shí)期雞腸道免疫細(xì)胞的發(fā)育情況,揭示雞腸道的免疫狀態(tài)和機(jī)制,為適時(shí)免疫,更好的防控禽病提供一定的參考資料。 通過(guò)HE染色方法研究雞回腸組織結(jié)構(gòu)變化情況,結(jié)果表明：1日齡時(shí),可見(jiàn)回腸絨毛呈指狀,腸腺較少；4日齡時(shí),絨毛高度同1日齡相比有所降低,腸絨毛固有層增寬,絨毛相互融合形成較大的皺襞,隱窩深度增加,在回腸黏膜固有層中可見(jiàn)有少量的淋巴濾泡；7日齡時(shí),絨毛高度增加,腸腺明顯,隱窩深度顯著增加(P0.05),超過(guò)其它日齡,達(dá)到峰值；10日齡時(shí),絨毛形狀多呈葉狀,絨毛高度、淋巴濾泡及杯狀細(xì)胞的增多；14日齡時(shí),隱窩深度到達(dá)穩(wěn)定水平；35日齡時(shí),絨毛高度到達(dá)穩(wěn)定水平。 通過(guò)免疫組織化學(xué)方法研究雞回腸中T淋巴細(xì)胞及其亞群的分布定位規(guī)律,結(jié)果表明：CD3+和CD8+細(xì)胞于18胚齡時(shí)出現(xiàn),CD4+于1日齡時(shí)出現(xiàn)。在分布定位上,CD3+細(xì)胞在回腸黏膜上皮及固有層中都有分布,CD4+以黏膜固有層中的分布為主,而CD8+T淋巴以黏膜上皮內(nèi)的分布為主。在數(shù)量變化上,4日齡時(shí),CD3+、CD4+及CD8+T淋巴細(xì)胞的數(shù)量均顯著增加；10日齡時(shí),CD3+和CD8+細(xì)胞的數(shù)量上升,CD4+細(xì)胞數(shù)量減少；14日齡時(shí),CD3+和CD8+細(xì)胞的數(shù)量減少,CD4+細(xì)胞數(shù)量增加；21日齡時(shí),CD3+、CD4+和CD8+T細(xì)胞數(shù)量顯著增加(P0.05),到達(dá)穩(wěn)定水平。 Bu-1+和IgA+細(xì)胞于18胚齡時(shí)出現(xiàn),IgG+細(xì)胞于1日齡時(shí)出現(xiàn)。在數(shù)量變化上,從1日齡到7日齡,Bu-1+細(xì)胞的數(shù)量一直保持增加的趨勢(shì)；10、14日齡時(shí),Bu-1+細(xì)胞的數(shù)量同7日齡相比略有下降；隨后,Bu-1+細(xì)胞的數(shù)量開(kāi)始增加,21日齡時(shí),Bu-1+細(xì)胞的數(shù)量同14日齡相比顯著增加(P0.05)；35日齡,Bu-1+細(xì)胞的數(shù)量到達(dá)峰值。IgA+細(xì)胞的數(shù)量變化整體呈逐漸增加的趨勢(shì)。18胚齡到10日齡,IgA+細(xì)胞的數(shù)量持續(xù)增加,21日齡時(shí),IgA+細(xì)胞的數(shù)量顯著增加,隨后,IgA+細(xì)胞的數(shù)量略有下降,但并不顯著。IgG+細(xì)胞在18胚齡到14日齡大致呈持續(xù)增加的趨勢(shì),僅在7日齡時(shí),IgG+細(xì)胞的數(shù)量稍有下降,但并不顯著。21日齡時(shí),IgG+細(xì)胞的數(shù)量顯著增加；35日齡時(shí),IgG+細(xì)胞的數(shù)量到達(dá)峰值；56日齡時(shí),IgG+細(xì)胞的數(shù)量同35日齡相比顯著下降(P0.05)。 腸道中淋巴細(xì)胞的發(fā)育,能夠促進(jìn)回腸組織結(jié)構(gòu)發(fā)育的完善,而腸道組織發(fā)育的一系列變化,為回腸相關(guān)免疫細(xì)胞的發(fā)育提供必須的微環(huán)境。這兩者互相協(xié)調(diào)作用,對(duì)機(jī)體的黏膜免疫系統(tǒng)的成熟起至關(guān)重要的作用。
[Abstract]:In this experiment, the fresh ileum of 10 chickens were collected from 18 ~ 20 embryo age chicken embryos and 1 ~ 4 ~ 4 ~ 7 ~ (10) ~ 10 ~ (14) ~ 21 ~ (-35) and 56 ~ (th) days old chicks respectively. The fresh ileum of 10 chickens were made into frozen sections and stained by HE and immunohistochemistry respectively. Based on image analysis and data statistics, the changes of tissue structure, the appearance, distribution and localization of lymphocytes in chicken ileum were studied. The aim of this study was to understand the development of intestinal immune cells in different periods and to reveal the immune status and mechanism of chicken intestinal tract so as to provide some reference materials for timely immunization and better prevention and control of poultry diseases. The changes of ileum tissue structure in chicken were studied by HE staining. The results showed that the ileal villi were finger shaped and the intestinal glands were less at 1 day of age. At 4 days of age, the height of villi decreased compared with that of 1 day, the lamina propria of intestinal villi widened, the villi fused to form larger folds, and the depth of crypt increased. A small number of lymphatic follicles could be seen in the lamina propria of ileum mucosa. At the age of 7 days, the villus height, intestinal gland and crypt depth increased significantly (P0.05), which reached the peak value over the other days, and at 10 days, the villi were mostly leaf shaped, villous height, lymphatic follicles and goblet cells increased. At the age of 14 days, the depth of crypt reached stable level, and the height of villi reached stable level at 35 days of age. The distribution and localization of T lymphocytes and their subsets in chicken ileum were studied by immunohistochemical method. The results showed that CD3 and CD8 cells appeared at 18 embryo age and CD4 appeared at 1 day old. In distribution localization, CD3 cells were distributed in ileal mucosa epithelium and lamina propria, CD4 was mainly distributed in lamina propria mucosa, and CD8 T lymphoid cells were mainly distributed in mucosal epithelium. The number of CD3, CD4 and CD8 T lymphocytes increased significantly at 4 days of age, and the number of CD3 and CD8 cells increased and the number of CD4 cells decreased at 10 days of age. At the age of 14 days, the number of CD3 and CD8 cells decreased and the number of CD4 cells increased, while at 21 days of age, the number of CD3, CD4 and CD8 T cells increased significantly (P0.05), reaching a stable level. Bu-1 and IgA cells appeared at 18 embryo age and IgG cells appeared at 1 day old. The number of Bu-1 cells increased from day 1 to day 7, and the number of Bu-1 cells decreased slightly at the age of 10 ~ 14 days. Then, the number of Bu-1 cells began to increase, and the number of Bu-1 cells increased significantly at 21 days old compared with 14 days old (P0.05). At the age of 35 days, the number of Bu-1 cells reached its peak. The number of IgA cells increased gradually. The number of IgA cells continued to increase from 18 to 10 days of age, and the number of IgA cells increased significantly at 21 days of age. The number of IgA cells decreased slightly, but not significantly. The number of IgG cells increased from 18 embryo to 14 days old, only at 7 days old, the number of IgG cells decreased slightly, but not significantly at 21 days old. The number of IgG cells increased significantly. At the age of 35 days, the number of IgG cells reached the peak; at the age of 56 days, the number of IgG cells decreased significantly compared with the age of 35 days (P0.05). The development of lymphocytes in the intestine can promote the development of the ileal tissue structure, and a series of changes of the intestinal tissue development provide the necessary microenvironment for the development of the ileal-associated immune cells. The coordination of the two plays an important role in the maturation of the mucosal immune system.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R392

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