Ueno Keigo, Urai Makoto, Ohkouchi Kayo, Miyazaki Yoshitsugu, Kinjo Yuki
Department of Chemotherapy and Mycoses, National Institute of Infectious Diseases, Tokyo, Japan.
Methods Mol Biol. 2016;1403:537-49. doi: 10.1007/978-1-4939-3387-7_30.
Several pathogenic fungi, including Cryptococcus gattii, Histoplasma capsulatum, Coccidioides immitis, and Penicillium marneffei, cause serious infectious diseases in immunocompetent humans. However, currently, prophylactic and therapeutic vaccines are not clinically used. In particular, C. gattii is an emerging pathogen and thus far protective immunity against this pathogen has not been well characterized. Experimental vaccines such as component and attenuated live vaccines have been used as tools to study protective immunity against fungal infection. Recently, we developed a dendritic cell (DC)-based vaccine to study protective immunity against pulmonary infection by highly virulent C. gattii strain R265 that was clinically isolated from bronchial washings of infected patients during the Vancouver Island outbreak. In this approach, bone marrow-derived DCs (BMDCs) are pulsed with heat-killed C. gattii and then transferred into mice prior to intratracheal infection. This DC vaccine significantly increases interleukin 17A (IL-17A)-, interferon gamma (IFN-γ)-, and tumor necrosis factor alpha (TNF-α)-producing T cells in the lungs and spleen and ameliorates the pathology, fungal burden, and mortality following C. gattii infection. This approach may result in the development of a new means of controlling lethal fungal infections. In this chapter, we describe the procedures of DC vaccine preparation and murine pulmonary infection model for analysis of immune response against C. gattii.
包括加氏隐球菌、荚膜组织胞浆菌、粗球孢子菌和马尔尼菲篮状菌在内的几种致病真菌可在免疫功能正常的人类中引发严重感染性疾病。然而，目前预防性和治疗性疫苗尚未在临床上使用。特别是，加氏隐球菌是一种新出现的病原体，迄今为止针对该病原体的保护性免疫尚未得到充分表征。诸如组分疫苗和减毒活疫苗等实验性疫苗已被用作研究针对真菌感染的保护性免疫的工具。最近，我们开发了一种基于树突状细胞（DC）的疫苗，以研究针对高毒力加氏隐球菌R265菌株肺部感染的保护性免疫，该菌株是在温哥华岛疫情期间从感染患者的支气管灌洗物中临床分离得到的。在这种方法中，用热灭活的加氏隐球菌对骨髓来源的DC（BMDC）进行脉冲处理，然后在气管内感染前将其转移到小鼠体内。这种DC疫苗可显著增加肺和脾中产生白细胞介素17A（IL-17A）、干扰素γ（IFN-γ）和肿瘤坏死因子α（TNF-α）的T细胞数量，并改善加氏隐球菌感染后的病理状况、真菌负荷和死亡率。这种方法可能会促成一种控制致命真菌感染的新方法的开发。在本章中，我们描述了DC疫苗制备程序以及用于分析针对加氏隐球菌免疫反应的小鼠肺部感染模型。
