Exosome-based tumor antigens-adjuvant co-delivery utilizing genetically engineered tumor cell-derived exosomes with immunostimulatory CpG DNA
Exosome-based tumor antigens-adjuvant co-delivery utilizing genetically engineered tumor cell-derived exosomes with immunostimulatory CpG DNA
复制标题
DOI：
10.1016/j.biomaterials.2016.09.031
复制
发表时间：
2016-12-01
期刊：
影响因子：
14
通讯作者：
Takakura, Yoshinobu
中科院分区：
文献类型：
--
作者：
Morishita, Masaki;Takahashi, Yuki;Takakura, Yoshinobu
对于通过肿瘤抗原疫苗接种与佐剂组合的癌症免疫疗法，主要挑战包括鉴定特定肿瘤抗原和将抗原以及佐剂有效递送至抗原呈递细胞。在这项研究中，我们提出了一种有效的基于外泌体的肿瘤抗原-佐剂共递送系统，使用基因工程肿瘤细胞来源的外泌体含有内源性肿瘤抗原和免疫刺激性CpG DNA。用编码融合链霉亲和素（SAV;以高亲和力结合生物素的蛋白质）-乳凝集素（LA;亲外泌体蛋白）蛋白的质粒载体转染鼠黑素瘤B16 BL 6细胞，产生基因工程改造的表达SAV-LA的外泌体（SAV-exo）。将SAV-exo与生物素化的CpG DNA结合制备CpG DNA修饰的外泌体（CpG-SAV-exo）。荧光显微镜观察显示，通过SAV-生物素相互作用，CpG DNA成功修饰了外泌体。CpG-SAV-exo显示出外泌体与CpG DNA有效且同时递送至培养的鼠树突状DC 2.4细胞。用CpG-SAV-exo处理有效地活化DC 2.4细胞并增强肿瘤抗原呈递能力。用CpG-SAV-exo免疫在B16 BL 6荷瘤小鼠中表现出比单纯共施用外泌体和CpG DNA更强的体内抗肿瘤作用。因此，基因工程CpG-SAV-exo是一种有效的基于外泌体的肿瘤抗原-佐剂共递送系统，其将用于癌症免疫治疗。(C)2016爱思唯尔有限公司版权所有
For cancer immunotherapy via tumor antigen vaccination in combination with an adjuvant, major challenges include the identification of a particular tumor antigen and efficient delivery of the antigen as well as adjuvant to antigen-presenting cells. In this study, we proposed an efficient exosome-based tumor antigens-adjuvant co-delivery system using genetically engineered tumor cell-derived exosomes containing endogenous tumor antigens and immunostimulatory CpG DNA. Murine melanoma B16BL6 cells were transfected with a plasmid vector encoding a fusion streptavidin (SAV; a protein that binds to biotin with high affinity)-lactadherin (LA; an exosome-tropic protein) protein, yielding genetically engineered SAV-LA-expressing exosomes (SAV-exo). SAV-exo were combined with biotinylated CpG DNA to prepare CpG DNA-modified exosomes (CpG-SAV-exo). Fluorescent microscopic observation revealed the successful modification of exosomes with CpG DNA by SAV-biotin interaction. CpG-SAV-exo showed efficient and simultaneous delivery of exosomes with CpG DNA to murine dendritic DC2.4 cells in culture. Treatment with CpG-SAV-exo effectively activated DC2.4 cells and enhanced tumor antigen presentation capacity. Immunization with CpG-SAV-exo exhibited stronger in vivo antitumor effects in B16BL6 tumor-bearing mice than simple co-administration of exosomes and CpG DNA. Thus, genetically engineered CpG-SAV-exo is an effective exosome-based tumor antigens-adjuvant co-delivery system that will be useful for cancer immunotherapy. (C) 2016 Elsevier Ltd. All rights reserved.