Roles of HSP60 in Helicobacter pylori infection

HSP60 在幽门螺杆菌感染中的作用

• 批准号: 12670256
• 负责人: YOKOTA Kenji
• 金额: $ 1.41万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670256/
• 关键词: Helicobacter phlori; Heat shock protein; Mucosa-associated lympho tissue (MALT); T helper 1 and 2 (Th-1 and -2); DNA microarray; Mucosal immunit; Heat shock protein; ヘリコバクターピロリ; HSP60; IgGサブクラス; Th-1 and Th-2; CD40L

We have previously reported that heat shock protein 60kDa (HSP60) is an important antigen in the pathogenesis of mucosa-associated lymphoid tissue (MALT) lymphoma. To investigate association with host immunity and HSP60, DC40 ligand (CD40L) and cytokine production in peripheral blood mononuclear cells (PBMCs) from the patients were analyzed following stimulation with HSP60. PBMCs obtained from patients with gastritis and MALT lymphoma, and those from healthy volunteer were stimulated with recombinant-HSP60 or H. pylori cell fysate in the presence of cytokines (IL-4) and GM-CSF). The mRNA expression was also analyzed by a cDNA microarray containing 1100 genes. The expression of CD40L on the CD4 positive cells was increased in the PBMCs from patients with MALT lymphoma stimulated by cytokines and/or HSP60 antigens. The production of IL-4 in PBMCs cultures was increased in patients with MALT lymphoma ; however, the production of IFN-_γ was at low levels. A cDNA microarray analysis indicat … More ed increased mRNA levels of HLA-DR and integrin. In cases of low-grade MALT lymphoma, adaptive immune responses against HSP60 may be enhanced by host factors, such as antigen presentation and T cell activation, resulting in B cell proliferation, which can be demonstrated during H. pylori infection.To study immunological roles of HSP60 in H. pylori infection, we immunized HSP60 to H. pylori infected mice. Whole HSP60 and two partial regions of helicobacterial hsp60 were expressed as GST-fusion proteins. Three recombinant proteins were designated rHSPw, rHSP2, and rHSP4-5. rHSPw was expressed as whole hsp60 (MeT_1-Met_<545>). rHSP2 (Glu_<101>-Ser_<200>) contained a domainT cell epitope cluster (Lys_<159>-Tnr_<178>). The rHSP4-5 contained a domain existing in both T cell epilope cluster (Asp_<396>-Gly_<4l2>) and its upstream of region (Ser_<356>-Asp_<392>) in common between H. pylori and human hsp60. Recombinant heat-labile enterotoxin (rLT) of E. coli was also employed as adjuvant for nasal immunization. Three different mouse groups (BALB/c, C3H/He, C57BL/6) were injected 10^8 CRJ of live H. pylori (SS -1) at two times a week and kept in a clean isolator for 2 weeks. Those mice were immunized H. pylori lysate or rHSPs (10μg/mouse) with rLT (10μg/mouse) by injecting into nasal mucosa four times in 1 month. Mice were sacrificed at 1 momtn after last immunization, and then histopathology and antibodies were investigated. Mucosal immunization by H. pylori lysate and rHSPs induced severe gastritis in BALB/c and C57BL/6 mice. Inflammation grades in those two groups immunized by rHSP4-5 were highest in aft groups. Mucosal immunization by hsp60 accelerated production of IgG and IgA.These -results indicated that HSP60 was closely associated with H. pylori induced gastric inflammation. Less

热休克蛋白60kDa（HSP 60）是粘膜相关淋巴组织（MALT）淋巴瘤发病机制中的一个重要抗原。为了研究与宿主免疫和HSP 60的相关性，分析了HSP 60刺激后患者外周血单核细胞（PBMC）中DC 40配体（CD40 L）和细胞因子的产生。用重组HSP 60或H.幽门螺杆菌细胞裂解物在细胞因子（IL-4）和GM-CSF的存在下）。用含有1100个基因的cDNA微阵列分析mRNA表达。MALT淋巴瘤患者PBMC经细胞因子和/或HSP 60抗原刺激后，CD4+细胞表面CD40L表达增加。MALT淋巴瘤患者PBMCs培养物中IL-4的产生增加，但IFN-γ的产生水平较低。cDNA微阵列分析表明， ...更多信息 艾德可增加HLA-DR和整合素的mRNA水平。在低度恶性MALT淋巴瘤的病例中，针对HSP 60的适应性免疫应答可能被宿主因素增强，例如抗原呈递和T细胞活化，导致B细胞增殖，这可以在H.研究HSP60在H. pylori感染中的免疫学作用。pylori感染后，用HSP60免疫H. pylori感染小鼠。整个HSP60和两个部分区域的螺旋杆菌hsp60表达GST融合蛋白。将三种重组蛋白命名为rHSP w、rHSP 2和rHSP 4 - 5。rHSPw表达为完整的hsp60（MeT_1-Met_）。<545>rHSP 2（Glu_-Ser_）含有一个结构域T细胞表位簇（Lys_-Tnr_）。<101><200><159><178>rHSP 4 - 5含有一个T细胞抗原簇结构域（Asp_-Gly_）及其上游区（Ser_-Asp_）。<396><4l2><356><392>pylori和人hsp60。重组大肠杆菌不耐热肠毒素（rLT）。大肠杆菌也被用作鼻免疫的佐剂。三个不同的小鼠组（BALB/c，C3H/He，C57BL/6）注射10^8 CRJ的活H。pylori（SS-1）感染，并在清洁隔离器中保存2周。这些小鼠免疫H. pylori裂解物或rHSPs（10 μ g/只）与rLT（10 μ g/只）联合鼻粘膜注射，共4次，1个月。末次免疫后1个月处死小鼠，观察组织病理学变化和抗体水平。用H. pylori裂解物和rHSPs诱导BALB/c和C57BL/6小鼠重度胃炎。rHSP 4 - 5免疫组和对照组的炎症分级均以aft组最高。HSP60免疫小鼠后，可促进IgG和IgA的产生，提示HSP60与H. pylori诱导的胃炎症。少

项目成果

Yunoki, N., Yokota. K., Mizuno, M., Kawahara, Y., Adachi, M., Okada, M., Hayashi, S., Hirai, Y., Oguma, K.,Tsuji, T.: "Antibody to heat shock protein can be used for early serological monitoring of Helicobacter pylori eradication treatment."Clin. Diag. La

Yunoki，N.，横田。

Hai-Xing Jiang, Hong Pu, Nam-Ho Huh, K.Yokota, et al.: "Helicobacter pylori induces pepsinogen secretion by rat gastric cells in culture via a cAMP signal pathway"Int.J.Molecul.Med.. 7. 625-629 (2001)

Hai-Xing Jiang、Hong Pu、Nam-Ho Huh、K.Yokota 等人：“幽门螺杆菌通过 cAMP 信号通路诱导培养物中大鼠胃细胞分泌胃蛋白酶原”Int.J.Molecul.Med.7。

Inaba, T., Mizuno, M., Kawai, K., Yokota K., Oguma, K., Miyoshi, M., Take, S., Okada, H., Tsuji, T: "Randomized open trial for comparison of proton pump inhibitors in triple therapy for Helicobacter pylori infection in relation to CYP2C19 genotype"J Gastr

Inaba, T.、Mizuno, M.、Kawai, K.、Yokota K.、Oguma, K.、Miyoshi, M.、Take, S.、Okada, H.、Tsuji, T：“随机公开试验，用于比较

Miyoshi, M., Mizuno, M., Ishiki, K., Nagahara, Y., Maga, T., Torigoe, T., Nasu, J., Okada, H., Yokota K., Oguma, K., Tsuji, T.: "A randomized open trial for comparison of proton pump inhibitors, omeprazole versus rabeprazole, in dual therapy for Helicobac

Miyoshi, M.、Mizuno, M.、Ishiki, K.、Nagahara, Y.、Maga, T.、Torigoe, T.、Nasu, J.、Okada, H.、Yokota K.、Oguma, K.、Tsuji

Miyoshi, M., Mizuno, M., Ishiki, K., Nagahara, Y., Maga, T., Torigoe, T., Nasu, J., Okada, H., Yokota K., Oguma K., Tsuji, T.: "A randomized open trial for comparison of proton pump inhibitors, omeprazole versus rabeprazole, in dual therapy for Helicobact

Miyoshi, M.、Mizuno, M.、Ishiki, K.、Nagahara, Y.、Maga, T.、Torigoe, T.、Nasu, J.、Okada, H.、Yokota K.、Oguma K.、Tsuji,