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Augmentation of cytokine production and mucosal immunity in gut-associated lymphoid tissue by the probiotics.

益生菌增强肠道相关淋巴组织中细胞因子的产生和粘膜免疫。

基本信息

批准号：
09660285
负责人：
KITAZAWA Haruki
金额：
$ 2.05万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

The Lactobacillus acidophilus group, a member of gastrointestinal lactic acid bacteria (LAB) widely used in dairy foods, is expected to develop as a probiotic LAB because of the ability to stimulate host immune responses. We have evaluated and assessed the mitogenic responses of the LAB to lymphocytes as an incidence of biological activity among them. The present study was conducted to determine the mitogenic factors from the LAB, particularly focusing on mitogenic DNAs in the LAB.In 12 of 16 strains of Lactobacillus acidophilus group LAB, the DNA components significantly stimulated mitogenic responses to murine splenocytes. The mitogenic activity to lymphocytes from peyer's patches, a gut-associated lymphoid tissue, was also observed in some DNAs. The active DNA (DNA1131) of Lactobacillus gasseri JCM1131^T was selected to analyze the active components. Plasmid DNAs from the positive clones obtained by suctioning, using the pUG119 vector ligated with 1131DNA partially digested with Sau … More 3Al, were purified by the precipitation method with polyethylene glycol. The plasmid DNA templates were amplified by using the polymerase chain reaction (POR) with primers for the multicloning site of the vector. In the 108 of the 321 positive colonies significant mitogenic activity was detected. The active nucleotides which stimulated mitogenic response were sequenced by a DNA sequencer MODEL 4000(Ll-COR, Lincoln, USA). Ten high homologous DNA motifs were found in the active DNA, but not in the non-active DNA.Two DNA motifs of chemically synthesized the DNA motifs induced a mitogenic response and was characterized as a B-cell specific mitogen. The binding of the active DNA clone and motifs to B lymphocytes were demonstrated by scatchard analysis and laser nicroscopy. The active DNA clone and motif augmented CD69/86 induction on B lymphocytes of spleen and peyer's patches. These also augmented induction of cytokine (IFNa. IFNy, TNRx) mRNA in splenocytes and peyer's patches. These findings indicated that a novel DNA motifs inducing lymphocyte activation triggering mitogenic response and , different from CpG motifs from E coli recently reported as a B-cell mitogen, exists in DNAs from L.Gasseri. Less

嗜酸乳杆菌是广泛用于乳制品中的胃肠道乳酸菌 (LAB) 的一员，由于能够刺激宿主免疫反应，预计将发展成为益生菌 LAB。我们评估并评估了 LAB 对淋巴细胞的促有丝分裂反应，作为其中生物活性的发生率。本研究旨在确定 LAB 中的促有丝分裂因子，特别关注 LAB 中的促有丝分裂 DNA。在 16 株嗜酸乳杆菌 LAB 组中的 12 株中，DNA 成分显着刺激了小鼠脾细胞的促有丝分裂反应。在一些 DNA 中也观察到了派尔氏淋巴结（一种肠道相关淋巴组织）淋巴细胞的有丝分裂活性。选取格氏乳杆菌JCM1131^T的活性DNA（DNA1131）进行活性成分分析。使用与Sau … More 3Al部分消化的1131DNA连接的pUG119载体，通过聚乙二醇沉淀法纯化抽吸获得的阳性克隆的质粒DNA。使用聚合酶链式反应 (POR) 和载体多克隆位点的引物扩增质粒 DNA 模板。在 321 个阳性集落中的 108 个中检测到显着的促有丝分裂活性。通过DNA测序仪MODEL 4000(L1-COR，Lincoln，USA)对刺激有丝分裂反应的活性核苷酸进行测序。在活性DNA中发现了10个高度同源的DNA基序，但在非活性DNA中没有发现。化学合成的DNA基序中的两个DNA基序诱导有丝分裂反应，并被表征为B细胞特异性有丝分裂原。活性DNA克隆和基序与B淋巴细胞的结合通过斯卡查德分析和激光显微镜证实。活性 DNA 克隆和基序增强了脾脏和集合淋巴结 B 淋巴细胞上的 CD69/86 诱导。这些还增强了脾细胞和集合淋巴结中细胞因子（IFNa、IFNy、TNRx）mRNA 的诱导。这些发现表明，L.Gasseri 的 DNA 中存在一种新的 DNA 基序，该基序诱导淋巴细胞激活，触发促有丝分裂反应，与最近报道的作为 B 细胞有丝分裂原的大肠杆菌 CpG 基序不同。较少的