Genome biology of staphylococcal enterotoxin family

葡萄球菌肠毒素家族的基因组生物学

• 批准号: 16380205
• 负责人: SHINAGAWA Kunihiro
• 金额: $ 9.34万
• 依托单位: Iwate University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16380205/
• 关键词: S.aureus; staphylococcal enterotoxin; recombinant DNA; genome biology; diagnosis; pathogenicity islands; plasmids; biological activities; pathogenicity island

In recent years, many new staphylococcal enterotoxins (SEs) have been reported. It has been known that certain SE genes are associated with mobile genetic elements such as pathogenicity islands, prophages, and plasmids. These facts imply that superantigenic toxin genes are transferred horizontally between staphylococcal strains. There is a possibility that these mobile genetic elements have played an important role in the evolution of S.aureus as a pathogen. To investigate pathobiology and epidemiology of these newly identified SEs, we had undertaken studies shown hi below.1.We have developed a comprehensive detection system for 18 kinds of classical and newly described staphylococcal superantigenic toxin genes using four sets of multiplex PCR. Analysis of the relationship between toxin genotypes and toxin genes encoding profiles of mobile genetic elements suggests its possible role in determining superantigenic toxin genotypes in S.aureus as combinations of toxin gene-encoding mobile … More genetic elements. Subsequently, we investigated new type of SE-encoding pathogenicity islands using PFGE/southern method. We have found that SEB-encoding islands are classified into at least 3 groups.2.We have determined complete nucleotides sequence of sed, selj and selj encoding plasmid, p196,using transposomics strategy. This plasmid is encoding penicillin resistance operon and Cd-resistance operon, in addition to three kinds of SE (SE1) genes.3.We characterized a novel staphylococcal entertoxin (SE)-like putative toxin SE1R and SE1P. The SE1R protein showed significant T cell stimulation activity. MHC class II molecules were required for T cell stimulation by SER. SER stimulated T cells bearing receptors Vβ 3,11,12,13.2, and 14. These results suggested that SER acts as a superantigen. SE1P induced a substantial proliferative response and the production of cytokines IL-2,EFN-γ,TNF-α, and IL-4 from human T cells when administered at a concentration of 0.4 pM or more. The expression of MHC class II molecules on accessory cells was required for T cell stimulation by SE1P. SE1P selectively stimulated a vast number of human T cells bearing receptors Vβ 5.1,6,8,16,18, and 21.3. These results indicated that SE1P acts as a superantigen. SE1P proved to be emetic in the house musk shrew emetic assay, although at a relatively high dose. We investigated whether immunization with non-toxic mutant SEC (mSEC), devoid of superantigenic activity, can protect against S.aureus infection. Immunization with mSEC devoid of superantigenic properties provides protection against S.aureus infection and the protection might be mediated by the IL-10 and IL-4 induction and down-regulation of IFN-γ, as well as SEC neutralizing antibodies. Also, we demonstrate that staphylococcal SEA induces an increase in intracellular calcium ([Ca^<2+>]i) in human intestinal epithelial cells and the [Ca^<2+>]i is released from intracellular stores. Less

近年来，许多新的葡萄球菌肠毒素（SE）被报道。已知某些SE基因与移动的遗传元件如致病岛、原噬菌体和质粒相关。这些事实表明，超抗原毒素基因在葡萄球菌菌株之间水平转移。这些移动的遗传元件可能在金黄色葡萄球菌作为病原体的进化中发挥了重要作用。为了研究这些新发现的金黄色葡萄球菌超抗原毒素的致病性和流行病学，我们进行了以下研究：1.建立了一套完整的检测体系，利用4套多重PCR技术检测18种经典和新发现的金黄色葡萄球菌超抗原毒素基因。对毒素基因型和编码移动的遗传元件的毒素基因谱之间的关系的分析表明，它在确定金黄色葡萄球菌中的超抗原毒素基因型中可能起作用，即编码移动的毒素基因的组合。 ...更多信息 遗传因素随后，我们研究了新型的SE编码致病岛使用PFGE/southern方法。2.利用转座组学方法测定了sed、selj和selj编码质粒p196的全序列。该质粒编码青霉素抗性操纵子和镉抗性操纵子，以及三种SE（SE 1）基因。3.我们鉴定了一种新的葡萄球菌肠毒素（SE）样推定毒素SE 1 R和SE 1 P。SE 1 R蛋白显示出显著的T细胞刺激活性。SER刺激携带Vβ 3、11、12、13.2和14受体的T细胞。这些结果表明SER是一种超抗原。当以0.4 pM或更高的浓度给药时，SE 1 P诱导了大量的增殖反应和人T细胞产生细胞因子IL-2、EFN-γ、TNF-α和IL-4。辅助细胞上MHC II类分子的表达是SE 1 P刺激T细胞所必需的。SE 1 P选择性刺激大量携带Vβ 5.1、6、8、16、18和21.3受体的人T细胞。这些结果表明，SE 1 P作为超抗原。SE 1 P被证明是在家麝鼩呕吐试验呕吐，虽然在一个相对较高的剂量。我们研究了用无超抗原活性的无毒突变体SEC（mSEC）免疫是否可以保护免受金黄色葡萄球菌感染。用缺乏超抗原特性的mSEC免疫提供针对金黄色葡萄球菌感染的保护，并且该保护可能由IL-10和IL-4诱导和IFN-γ的下调以及SEC中和抗体介导。此外，我们还证实了金黄色葡萄球菌SEA可诱导人肠上皮细胞内钙离子（[Ca^2+]i）的增加，并且[Ca^2+]i从细胞内储存中释放出来。少

项目成果

Staphylococcal enterotoxin A modulates intracellular Ca2+ signal pathway in human intestinal epithelial cells

• DOI: 10.1016/j.febslet.2005.07.005
• 发表时间: 2005-08-15
• 期刊: FEBS LETTERS
• 影响因子: 3.5
• 作者: Hu, DL;Suga, S;Nakane, A
• 通讯作者: Nakane, A

Biological Properties of Staphylococcal Enterotoxin-Like Toxin Type R

• DOI: 10.1128/iai.72.6.3664-3667.2004
• 发表时间: 2004-06
• 期刊: Infection and Immunity
• 影响因子: 3.1
• 作者: K. Omoe;K. Imanishi;D. Hu;H. Kato;H. Takahashi-Omoe;A. Nakane;T. Uchiyama;K. Shinagawa

Characterization of novel staphylococcal enterotoxin-like toxin type P

• DOI: 10.1128/iai.73.9.5540-5546.2005
• 发表时间: 2005-09-01
• 期刊: INFECTION AND IMMUNITY
• 影响因子: 3.1
• 作者: Omoe, K;Imanishi, K;Shinagawa, K
• 通讯作者: Shinagawa, K

A mutant of staphylococcal enterotoxin C devoid of bacterial superantigenic activity elicits a Th2 immune response for protection against Staphylococcus aureus infection

• DOI: 10.1128/iai.73.1.174-180.2005
• 发表时间: 2005-01-01
• 期刊: INFECTION AND IMMUNITY
• 影响因子: 3.1
• 作者: Hu, DL;Cui, JC;Nakane, A
• 通讯作者: Nakane, A

Comprehensive analysis of classical and newly described staphylococcal superantigenic toxin genes in Staphylococcus aureus isolates

• DOI: 10.1016/j.femsle.2005.04.007
• 发表时间: 2005-05-15
• 期刊: FEMS MICROBIOLOGY LETTERS
• 影响因子: 2.1
• 作者: Omoe, K;Hu, DL;Shinagawa, K
• 通讯作者: Shinagawa, K