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研究生:白欣怡
研究生(外文):Hsin-Yi Pai
論文名稱:熱致死乳酸桿菌促使老鼠脾臟細胞及經樹突細胞刺激之T細胞的免疫反應偏向Th1
論文名稱(外文):Heat-killed cells of lactobacilli skew the immune response toward T helper 1 polarization in mouse splenocytes and dendritic cell-treated T cells
指導教授:林美吟林美吟引用關係
指導教授(外文):Meei-Yn Lin
學位類別:碩士
校院名稱:國立中興大學
系所名稱:食品暨應用生物科技學系
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:77
中文關鍵詞:乳酸桿菌脾臟細胞樹突細胞第一型輔助T細胞細胞激素
外文關鍵詞:Lactobacillussplenocytesdendritic cellsT helper 1 cellscytokines
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乳酸菌為人類及哺乳類腸胃道中的共生細菌之一,已被認定為益生菌,對於維持宿主健康扮演著重要的角色,包括了其對於免疫功能的調節。目前大部分的研究皆比較著重於乳酸菌的活菌體 (viable cell) 及其細胞壁萃取物 (cell wall extract) 對於免疫調節作用的影響,而本實驗則主要是在於乳酸菌熱致死菌體的影響。我們藉由將三株熱致死乳酸菌L. acidophilus A2、L. gasseri A5和L. salivarius A6與脾臟細胞共同培養,來探討此三株熱致死乳酸桿菌對於免疫功能所產生的影響。另一方面,我們亦將三株熱致死乳酸菌L. acidophilus A2、L. gasseri A5和L. salivarius A6刺激樹突細胞後,再將經過刺激的樹突細胞與T細胞共同培養,觀察T細胞的免疫反應。由實驗結果顯示三株熱致死乳酸菌L. acidophilus A2、L. gasseri A5和L. salivarius A6,證實能刺激BALB/c小鼠的脾臟細胞增生和interleukin (IL)-10、IL-12 p70和interferon (IFN)-γ之分泌,但對transforming growth factor (TGF)-β無影響,使免疫反應偏向Th1型免疫反應。另外,三株熱致死乳酸菌亦能活化BALB/c小鼠的樹突細胞分泌大量的IL-12 p70,且經過刺激的樹突細胞能夠促使T細胞活化增生並顯著提升Th1型的細胞激素IFN-γ之分泌,但對IL-5, IL-13及TGF-β並無影響。以上結果顯示乳酸桿菌 (lactobacilli) 對於調節免疫機能及過敏反應扮演了重要的角色。
It is believed that probiotics play an important role for the health of the host including modulation of immune responses. Most studies have focused on the immunomodulatory effects of viable cells of lactic acid bacteria, however we investigated those of heat-killed cells of lactic acid bacteria in this study. We first observed the effects on immune functions via stimulating splenocytes with three heat-killed Lactobacillus strains. Furthermore, we also investigated the effect of mouse dendritic cells (DCs) treated with these heat-killed Lactobacillus strains on T cell responses. The results showed that these Lactobacillus strains were able to stimulate cell proliferation and interleukin (IL)-10, IL-12 p70 and interferon (IFN)-γ production, but not transforming growth factor (TGF)-β in splenocytes. In addition, these heat-killed Lactobacillus strains also stimulated high-level secretion of IL-12 p70 in DCs and switched T cells to T helper (Th) 1 immune responses, as evidenced by the elevated secretion of IFN-γ, but not IL-5, IL-13 and TGF-β. These results showed that lactobacilli play a potentially important role in modulating immune responses and allergic reactions.
目 次

中文摘要...........................................i
Abstract..........................................ii
目次.............................................iii
表目次............................................vi
圖目次...........................................vii

第一章 前言.......................................1

第二章 文獻整理...................................3
一、乳酸菌之概述...................................3
二、乳酸菌之益生功能...............................3
(一) 維持腸道內菌相平衡............................4
(二) 減緩乳糖不耐症................................5
(三) 降低膽固醇....................................5
(四) 預防癌症......................................5
(五) 調節免疫系統..................................6
(六) 降低血壓......................................7
三、免疫系統之概述.................................7
(一) 先天性免疫 (Innate immunity)..................7
(二) 適應性免疫 (Adaptive immunity)................9
四、樹突細胞之功能................................10
五、T細胞之功能...................................11
(一) 免疫細胞與細胞激素...........................11
(二) Th1與Th2免疫細胞............................12
六、腸道黏膜免疫系統..............................14
(一)免疫系統之器官................................14
(二) 腸道相關淋巴組織 (GALT)......................14
(三) 腸道內的免疫反應.............................15
七、乳酸菌免疫調節作用............................17

第三章 實驗目的..................................26
一、目的..........................................26
二、實驗架構......................................27

第四章 材料與方法................................28
一、材料..........................................28
(一) 試驗菌株.....................................28
(二) 實驗動物.....................................28
(三) 實驗器材及設備...............................28
(四) 實驗藥品.....................................29
(五) 培養基與緩衝溶液之配製.......................31
二、方法..........................................33
(一) 乳酸菌之製備.................................33
(二) 樹突細胞之製備...............................34
(三) 脾臟細胞及T細胞之製備........................34
(四) 脾臟細胞免疫刺激試驗.........................36
(五) 樹突細胞免疫刺激試驗.........................36
(六) T細胞免疫刺激試驗...........................36
(七) 細胞激素之測定...............................37
(八) 細胞增生之測定...............................38
(九) 統計分析.....................................39

第五章 結果與討論................................40
一、結果..........................................40
(一) 乳酸菌對脾臟細胞增生之影響...................40
(二) 乳酸菌對脾臟細胞分泌細胞激素之影響...........40
(三) 乳酸菌對樹突細胞分泌細胞激素之影響...........42
(四) 乳酸菌對T細胞增生之影響......................43
(五) 乳酸菌對T細胞分泌細胞激素之影響..............43
二、討論..........................................45
(一) 乳酸菌對脾臟細胞之影響.......................45
(二) 乳酸菌對樹突細胞及T細胞之影響................47
三、結論..........................................49

第六章 參考文獻...................................63
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