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標題: | 多重抗藥性K62及K64克雷伯氏肺炎桿菌莢膜接合疫苗在免疫功能不全小鼠模式之效用評估 Efficacy of K62 and K64 Capsule Conjugate Vaccines of Multidrug Resistant Klebsiella pneumoniae in Immunocompromised Mice |
作者: | 柯彥溥 Yen-Pu Ko |
指導教授: | 王錦堂 Jin-Town Wang |
關鍵字: | 克雷伯氏肺炎桿菌,多重抗藥性,莢膜分解酵素,莢膜接合疫苗, Klebsiella pneumoniae,multidrug resistant,capsule depolymerase,capsule conjugate vaccine, |
出版年 : | 2018 |
學位: | 碩士 |
摘要: | 克雷伯氏肺炎桿菌(Klebsiella pneumoniae)是人類呼吸道及消化道的正常菌叢,會伺機性造成免疫不全病人之院內感染及正常人之社區型感染。多重抗藥性克雷伯氏肺炎桿菌更是造成臨床上治療病人極大的困難,利用疫苗使宿主的免疫系統產生記憶性抗體作為替代療法可能是一個不錯的方法。先前實驗室研究台大、成大、榮總及長庚四家醫院之碳青黴烯類抗生素抗藥性的克雷伯氏肺炎桿菌臨床菌株之莢膜型,其中5種莢膜型(K64、K62、K24、KN2、K28)就佔了約72%的菌株。首先,本研究成功表現並純化出K24及K28莢膜分解酵素,分別針對K24及K28菌株有良好的活性。接著利用先前實驗室所純化出的噬菌體莢膜分解酵素將細菌莢膜切割成小片段醣分子,並與易誘發免疫反應的載體蛋白質CRM197連接製成莢膜接合疫苗,評估兩個多重抗藥性菌株最常見莢膜型K62及K64之莢膜接合疫苗在BALB/c小鼠之安全性、抗體產生能力,及以cyclophosphamide建立免疫不全小鼠模式模擬疫苗是否可對臨床上免疫不全病人感染多重抗藥性克雷伯氏肺炎桿菌提供保護力。K62及K64莢膜接合疫苗安全性方面,小鼠接種疫苗18-24小時後量測肛溫及體重並無不良反應。K62莢膜接合疫苗可誘發K62莢膜抗體產生,其血清殺菌試驗約有40%(6/15)小鼠血清為陽性反應,接種四劑及五劑K62莢膜接合疫苗可顯著增加感染致死劑量之K62菌株的免疫抑制小鼠存活率。K64莢膜接合疫苗誘發抗體產生能力不佳,推測可能由於K64菌株之莢膜結構抗原性差或是接合的方法等原因所造成。 Klebsiella pneumoniae is normal flora of human respiratory tract and digestive tract, and sometimes becomes opportunistic pathogen which causes nosocomial infection of immunocompromised patients or community-acquired infection of healthy people. Multidrug resistant K. pneumoniae(MDRKP) is a clinical urgent problem. Preventive immunization through vaccines may be an alternative treatment against MDRKP infection in prospective host. Our previous study showed that K64, K62, K24, KN2, and K28 capsular type account for 72% of carbapenem-resistant K. pneumoniae (CRKP) collected from 4 hospitals of Taiwan. In the first part of this study, K24 and K28 capsule depolymerases were successfully expressed and purified, and the enzymatic activity toward K24 and K28 strains was demonstrated, respectively. Next, we used previously purified capsule depolymerases to digest capsule into fragments, which were conjugated with high immunogenic carrier protein CRM197 to generate capsule conjugate vaccines. K62 and K64 capsule conjugate vaccines, two of the CRKP prevalent capsular type, were generated and evaluated. The safety, antibody production ability, serum bactericidal activity, and protective efficacy in cyclophosphamide-treated BALB/c mice, which mimic immunocompromised patients, were studied. 18-24 hours after immunization, no adverse effect was observed in body temperature and weight of K62 and K64 conjugate vaccine-immunized mice. K62 conjugate vaccine induced the production of K62 capsule antibody, and provoked the serum bactericidal activity in 6/15 (40%) of mice. Receiving 4 or 5 doses of K62 conjugate vaccine significantly increased the survival of immunocompromised mice when challenged with lethal dose of K62 strain. However, K64 conjugate vaccine failed to induce the production of K64 capsule antibody. We speculate low antigenicity of K64 capsular polysaccharide structure or conjugation methods might affect the immunogenicity of K64 capsule conjugate vaccine. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77802 |
DOI: | 10.6342/NTU201801354 |
全文授權: | 未授權 |
電子全文公開日期: | 2023-10-11 |
顯示於系所單位: | 微生物學科所 |
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