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，然而，它们患有慢性肠道疾病，导致粮食产量下降和抗生素过量使用，这成为非常严重的问题。本研究的最终目标是开发一种新型口服疫苗佐剂。为了实现最终目标，我们首先尝试从乳酸和细菌（LAB）中寻找活性DNA基序，并利用猪TLR9转染子开发新的免疫分析系统。阐明 LAB DNA 在肠道中的免疫刺激机制我们发现了多种免疫刺激 DNA 基序，其中包括来自 LAB 的一种新基序我们从猪派尔氏淋巴结中克隆了 Toll 样受体 9 (TLR9)，这预计将用于人类模型。并成功开发了猪 TLR9 转染子，该转染子可用于评估 LAB 的活性 DNA。DNA 基序增强了 Th1 型细胞因子的诱导。 (IL-6, IL-12, IFNγ) mRNA 在派伊尔淋巴结中表达猪 TLR9 基因优先在派伊尔淋巴结 (Pps) 和肠系膜淋巴结 (MLN) 中表达，其表达程度约为脾脏的 3 倍。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。 Pps 和 MLN 不仅存在于树突状细胞和 B 细胞等免疫细胞中，还存在于 Pps 中属于滤泡相关上皮 (FAE) 的膜细胞 (M 细胞) 中。从这些结果来看，来自 LAB 的免疫刺激 DNA 基序通过 TLR9 在肠道中发挥细胞因子诱导作用。表明 LAB DNA 是口服疫苗佐剂的良好候选者。