本文選題：咪喹莫特　切入點(diǎn)：疫苗　出處：《中國(guó)協(xié)和醫(yī)科大學(xué)》2007年碩士論文

【摘要】： 皮膚是人體的最大的組織和器官，它的表面積接近2m~2。皮膚也是一個(gè)具有獨(dú)特免疫功能并與全身免疫系統(tǒng)密切相關(guān)的組織器官。 德國(guó)的醫(yī)學(xué)學(xué)生Langerhans P于1868年首次發(fā)現(xiàn)并描述了皮膚中的抗原遞呈細(xì)胞，后被命名為朗格漢斯細(xì)胞(Langerhans cell，LC)。未成熟的朗格漢斯細(xì)胞具有較強(qiáng)吞噬處理抗原能力，并沿淋巴管遷移至近側(cè)引流淋巴結(jié)(draining lymph nodes，DLN)的T細(xì)胞區(qū)并分化成熟而具有很強(qiáng)的抗原遞呈(包括交叉遞呈)能力。 透皮免疫(Transcutaneous immunization，TCI)是一種新的疫苗接種方法。它通過(guò)在皮膚表面局部應(yīng)用免疫佐劑和疫苗抗原來(lái)使機(jī)體產(chǎn)生全身性免疫反應(yīng)。通過(guò)透皮劑等技術(shù)手段，使疫苗抗原進(jìn)入表皮中并被表皮朗格漢斯細(xì)胞吞噬，在佐劑的作用下，迅速引發(fā)全身性的免疫反應(yīng)，從而達(dá)到免疫效果。 2004年Karande等發(fā)現(xiàn)了透皮劑聚氧乙烯十二烷基醚硫酸鈉(sodium laureth sulfate，SLA)+苯基哌嗪(phenyl piperazine，PP)的組合，該組合能夠有效促進(jìn)分子量小于1～10kD的生物大分子的透皮，并不誘導(dǎo)皮膚損傷。SLA+PP的組合能夠在離體的皮膚中最大限度的促進(jìn)肝素、促黃體(生成)激素-釋放激素等生物大分子的透皮；在小鼠皮膚上的實(shí)驗(yàn)也證實(shí)醋酸亮丙瑞林能夠在促透劑的作用下進(jìn)入小鼠體內(nèi)。 在研究人類單純皰疹病毒的治療時(shí)發(fā)現(xiàn)imiquimod具有很強(qiáng)的抗病毒和抗腫瘤活性，美國(guó)3M公司在1997年將其上市，制成5％的外用軟膏劑(商品名Aldara)。咪喹莫特可以刺激單核細(xì)胞、巨噬細(xì)胞、樹突狀細(xì)胞產(chǎn)生以干擾素IFN-α、腫瘤壞死因子TNF-α、白介素IL-1、6、8、10、12為代表的多種細(xì)胞因子。我們用咪喹莫特免疫刺激特性作為透皮佐劑，研究亞單位疫苗、病毒疫苗、DNA疫苗三種疫苗的透皮免疫效果。 聚氧乙烯十二烷基醚硫酸鈉和苯基哌嗪混合之后溶于等體積PBS緩沖液和無(wú)水乙醇混合液中，，制成透皮劑。將其分別與HBV亞單位抗原、HBsAg DNA疫苗、滅活的HAV抗原混合后，涂抹于小鼠脫毛的背部皮膚表面，同時(shí)涂抹咪喹莫特作為佐劑，分別在2、4、6、8、12、20周用ELISA法檢測(cè)血清中的抗體水平。在透皮免疫2周后即可在用咪喹莫特作為佐劑的HBV亞單位抗原組和DNA疫苗組中用ELISA法檢測(cè)到血清中的抗體，平均分別為1：19.1和1：30.3；抗體滴度在8周達(dá)到最高水平，平均分別為1：105.6和1：105.6；無(wú)佐劑組在4周才檢測(cè)到抗體，抗體滴度平均為1：3.31和1：1.82。結(jié)論：咪喹莫特能高效迅速誘導(dǎo)HBV亞單位抗原和DNA疫苗透皮免疫的抗體產(chǎn)生。
[Abstract]:Skin is the largest tissue and organ of the human body, and its surface area is close to 2 mm2. The skin is also a tissue and organ with unique immune function and closely related to the systemic immune system. Langerhans P, a German medical student, first discovered and described antigen-presenting cells in the skin in 1868, and was later named Langerhans cells. The immature Langerhans cells had a strong phagocytic ability to process antigens. Along the lymphatic vessels, they migrated to the T cell region of the proximal draining lymph nodes-DLNs and differentiated and matured and had strong antigen-presenting (including cross-presentation) capacity. Transdermal immunity (TCI) is a new vaccination method. It can induce systemic immune response by local application of immune adjuvant and vaccine antigen on skin surface. The vaccine antigen enters the epidermis and is swallowed by the Langerhans cells of the epidermis. With the action of adjuvant, the immune response of the whole body is triggered quickly, and the immune effect is achieved. In 2004, Karande et al discovered the combination of sodium laureth sulfate (sodium laureth sulfate) phenyl piperazine (PPP), which can effectively promote the transdermal penetration of biological macromolecules with molecular weight less than 1~10kD. The combination of SLAPP can promote the transdermal penetration of heparin, luteinizing hormone (luteinizing) hormone and other biological macromolecules in vitro. Experiments on the skin of mice also confirmed that Leuprilline acetate was able to enter the mice under the action of a penetration enhancer. When studying the treatment of human herpes simplex virus (HSV), we found that imiquimod has strong antiviral and antitumor activity. Us 3M Company went on the market in 1997 and made 5% external ointment (Aldaraan.imiquimod) to stimulate monocytes. Macrophages, dendritic cells produce a variety of cytokines represented by IFN- 偽, TNF- 偽, IL-1TNF- 偽, IL-1TNF- 偽 and IL-1. We use the immunostimulatory properties of Imiquimod as transdermal adjuvants to study subunit vaccines. The transdermal immune effect of three kinds of virus vaccine and DNA vaccine. Sodium polyoxyethylene dodecyl ether sulfate and phenylpiperazine were dissolved in the mixture of PBS buffer and anhydrous ethanol to prepare transdermal agent. The transdermal agent was mixed with HBV subunit antigen, DNA vaccine, and inactivated HAV antigen. Apply to the skin surface of the mouse's back with imiquimod as adjuvant. The antibody levels in serum were detected by ELISA method at the week of 20 weeks, respectively. After 2 weeks of transdermal immunization, the antibodies in serum were detected by ELISA method in the HBV subunit antigen group and DNA vaccine group with imiquimod as adjuvant. The antibody titer reached its highest level at 8 weeks, with an average of 1: 105.6 and 1: 105.6, respectively; the adjuvant group did not detect antibodies until 4 weeks. The average titer of antibody was 1: 3.31 and 1: 1.82.Conclusion: Imiquimod can efficiently and rapidly induce the production of HBV subunit antigen and DNA vaccine transdermal antibody.
【學(xué)位授予單位】：中國(guó)協(xié)和醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R392

【共引文獻(xiàn)】

相關(guān)期刊論文 前3條

1 楊淑靜;霍亂毒素[J];生物技術(shù)通訊;2000年04期

2 陳菁;彭艷萍;李玉良;陳俊;王秋根;;CO_2激光聯(lián)合咪喹莫特乳膏治療女性尖銳濕疣療效觀察[J];臨床醫(yī)藥實(shí)踐;2009年20期

3 荊慧琴;蘇玩琴;白云;;咪喹莫特乳膏治療尖銳濕疣臨床療效觀察[J];實(shí)用醫(yī)技雜志;2006年21期

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