Oral vaccine from daily lactic acid bacteria! -Development of new vaccine adjuvants-

每日乳酸菌口服疫苗！

基本信息

批准号：
13556018
负责人：
KITAZAWA Haruki
金额：
$ 8.83万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13556018/
关键词：
Lactic acid bacteria Immunobiotics L.gasseri DNA Peyer's patch TLR9 Transfectant Cytokine L.gasseri 腸管膜リンパ節 L. bularicus トランスフェクタン S.thermophilus CpG motif 腸管クローニング

项目摘要

Domestic animals are very important for food production. However, it becomes very severe problem that they have chronic intestinal diseases resulted in loss of food production and overadministration of antibiotics. In this study, the final goal is development of a novel oral vaccine adjuvant by using dairy lactic and bacterial DNA. In order to go ahead to the final goal, we first tried to find the active DNA motifs from lactic and bacteria (LAB) and develop a new immunoassay system with swine TLR9 transfectant and finally elucidate the mechanisms of the immunostimulation by the LAB DNA in the gut. We found several immunostimulatory DNA motifs including a novel one from the LAB. We cloned Toll-like receptor 9 (TLR9) from swine peyer's patch, which is expected for human model, and successfully developed a swine TLR9 transfectant. The transfectant was useful for the evaluation of active DNA from the LAB. The DNA motifs augmented an induction of Th1 type cytokine (IL-6, IL-12, IFNγ) mRNA in peyer's patches. The swine TLR9 gene was preferentially expressed peyer's patches (Pps) and mesenteric lymph nodes (MLNs), of which their degrees were approximately three times higher than that of spleens but weak in the other tissues. We firstly showed that high express (TLR9^+) cells were formed in Pps and MLNs. In addition, TLR9^+ cells existed not only in immune cells such as dendritic cells and B cells but also in membranous cells (M cells), belonging to follicle-associated epithelia (FAE) in Pps. These results suggest that Pps and MLNs provide the host defense with the ability to respond to a variety of bioactive oligonucleotides (ODNs) from bacteria at a conductive site of initial immune responses. From these results, immunostimulatory DNA motif from the LAB exerted the cytokine induction in the gut via TLR9 suggesting that the LAB DNA is a good candidate of an oral vaccine adjuvant.

家畜对于粮食生产非常重要。但是，他们患有慢性肠道疾病会导致食品生产和抗生素过度给药变得非常严重。在这项研究中，最终目标是通过使用乳酸乳酸和细菌DNA的新型口服疫苗的开发。为了实现最终目标，我们首先试图从乳酸和细菌（实验室）找到活跃的DNA图案，并开发具有猪TLR9透明的新免疫系统，并最终阐明了肠道中实验室DNA免疫刺激的机制。我们发现了几个免疫刺激性DNA基序，包括实验室的新型DNA基序。我们从猪佩耶（Swine Peyer）的斑块中克隆了类似Toll的受体9（TLR9），该贴片是人类模型的预期，并成功地开发了猪TLR9透明。转化剂可用于评估实验室的活性DNA。 DNA基序增强了Peyer斑块中Th1型细胞因子（IL-6，IL-12，IFNγ）mRNA的指示。猪TLR9基因优选为Peyer的斑块（PPS）和肠系膜淋巴结（MLN），其学位比脾脏高约三倍，但在其他组织中则较弱。我们首先表明在PPS和MLN中形成了高表达（TLR9^+）细胞。此外，TLR9^+细胞不仅存在于诸如树突状细胞和B细胞等免疫细胞中，而且还存在于PPS中属于卵泡相关的上皮（FAE）的膜细胞（M细胞）中。这些结果表明，PPS和MLN提供了宿主的防御能力，能够对来自初始免疫调查的导电部位的细菌的多种生物活性寡核苷酸（ODN）做出反应。从这些结果中，实验室的免疫刺激DNA基序通过TLR9在肠道中施加了细胞因子诱导，这表明实验室DNA是口服疫苗调整的良好候选者。