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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

嗜酸乳杆菌群是广泛应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，因此有望发展为益生LAB。我们已经评价和评估了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。本研究的目的是确定从乳酸菌的促有丝分裂因子，特别是专注于有丝分裂DNA在LAB。在12的16株嗜酸乳杆菌组LAB，DNA成分显着刺激小鼠脾细胞的促有丝分裂反应。在某些DNA中还观察到对来自派伊尔集合淋巴结（一种肠道相关淋巴组织）的淋巴细胞的促有丝分裂活性。以加氏乳杆菌JCM 1131 ^T的活性DNA（DNA 1131）为研究对象，对该菌株的活性成分进行了分析。使用与用Sau部分消化的1131 DNA连接的pUG 119载体，通过抽吸获得来自阳性克隆的质粒DNA ...更多信息 3 Al，用聚乙二醇沉淀法提纯。质粒DNA模板通过使用聚合酶链反应（POR）用针对载体的多克隆位点的引物扩增。在321个阳性集落中的108个中检测到显著的促有丝分裂活性。通过DNA测序仪MODEL 4000（L1-COR，林肯，美国）对刺激促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模体，而在非活性DNA中没有发现，其中两个DNA模体能诱导有丝分裂反应，是B细胞特异性的有丝分裂原。Scatchard分析和激光显微镜观察证实了活性DNA克隆和基序与B淋巴细胞的结合。活性DNA克隆和基序增强脾和派伊尔结B淋巴细胞上的CD 69/86诱导。这些也增强了细胞因子（IFNa.脾细胞和派伊尔集合淋巴结中IFN γ、TNRx）mRNA。这些结果表明，一个新的DNA基序诱导淋巴细胞活化触发有丝分裂反应，不同的CpG基序从大肠杆菌最近报道的B细胞有丝分裂原，存在于L. GasseriDNA。少