α-galactosylceramide衍生物嵌合微脂粒對樹突狀細胞的成熟影響與點鼻給予之免疫效應

Study on liposomes incorporating α-galactosylceramide analogs in the induction of dendritic cells maturation and intranasal immunological effect

江珮慈(Pei-Tzu Chiang)

指導教授 ： 劉得任

臺北醫學大學/口腔醫學院/生醫材料暨工程研究所/碩士(2011年)

https://doi.org/10.6831/TMU.2011.00087

摘要

黏膜免疫系統在抵禦外來病原體提供極重要的功能；在執行防禦功能時，樹突狀細胞在抗原呈現上扮演重要的角色。α-Galactosylceramide (α-GalCer)是由海綿分離出來的天然物，已知其可透過抗原呈現細胞(antigen-presenting-cells, APCs)上的CD1d分子媒介，活化Natural Killer T (NKT)細胞，並產生大量Th1和Th2細胞激素(cytokines) ，進而活化各種先天性(innate)及後天性(adaptive)免疫細胞產生免疫力。
本實驗利用α-GalCer類似物與PC (phosphatidylcholine)製成之中性電荷微脂粒，在細胞實驗觀察樹突狀細胞發育過程中，給予嵌合有α-GalCer類似物的微脂粒及傳統無嵌合式微脂粒， 觀察其對樹突狀細胞表面分子CD80及CD86表現量變化影響，結果顯示微脂粒組別均有增加的趨勢；為了瞭解嵌合式微脂粒經點鼻給予的免疫效應，本實驗以六週齡BALB/c母鼠，經鼻黏膜免疫二次接種後，收集血清、鼻沖洗及肺沖洗液，以酵素連結免疫分析法(ELISA)測量其IgA、IgG及subtype IgG生成量來了解嵌合式微脂粒與傳統微脂粒在免疫形成的影響，結果顯示。經嵌合有α-GalCer及其類似物之微脂粒二次點鼻免疫能誘使更高的黏膜分泌型免疫球蛋白sIgA及血清免疫球蛋白IgG生成。
由細胞實驗結果證實，微脂粒良好的載體特性確實能誘使樹突狀細胞成熟；而嵌合有α-GalCer各組別之微脂粒在二次點鼻免疫後兩週，能比傳統微脂粒於呼吸道黏膜誘導產生更大量的保護性抗體IgA及增加血液中具全身保護性抗體IgG的產生。

關鍵字

微脂粒 ； 黏膜免疫 ； alpha-Galcer

並列摘要

The mucosal immune systems serve vital functions to against foreign pathogens. In performing these functions, mucosal dendritic cells (DCs) plays an important role of antigen presenting. α-Galactosylceramide (α-GalCer) is presented by CD1d molecule on APCs to invariant NKT (iNKT) cells, which produced rapidly large amounts of Th1 and Th2 cytokines upon activation, leading to activation of a variety of innate and adaptive immune cells.
We incorporated α-GalCer to PC (phosphatidylcholine) and formed neutral charge of liposome. In this study, we firstly investigated whether liposome incorporated with α-GalCer analogs played an immune-modulatory role in the activation and function of DCs in vitro. The DCs were treated with liposome incorporated with α-GalCer analogs and traditional liposome during development of cells. The results showed that the levels of CD80 and CD86 expression were higher presence in all of liposomal groups. In vivo, female BALB/c mice (6-weeks-old) were immunized by intranasal route with α-GalCer incorporated liposome, compared with traditional liposome. After immunization, mice were sacrificed to collect the serum, nasal and lung washes. We analyzed the IgA, IgG, and subtype IgG production by enzyme-linked immunosorbent assays (ELISA). We found that intranasal immunization with α-GalCer and its analogs incorporated liposomes elicit higher mucosal secretory immunoglobulin A (sIgA) and serum IgG production.
In vitro, the maturation of dendritic cells confirmed the effectiveness of delivered ability of liposome. In vivo, animal experiment showed that α-GalCer incorporated liposome induced significantly increased levels of IgA responses in mucosal compartments and also enhancing IgG levels better than traditional liposomes.

並列關鍵字

liposome ； mucosal immunity ； alpha-Galcer

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