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; 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

我们先前已经报道说，热休克蛋白60KDA（HSP60）是粘膜相关淋巴组织（MALT）淋巴瘤发病机理中的重要抗原。为了研究与HSP60刺激后，分析了患者外周血单核细胞（PBMC）中与宿主免疫和HSP60的关联，DC40配体（CD40L）和细胞因子产生的关联。在存在细胞因子（IL-4）和GM-CSF的情况下，从胃炎和麦芽淋巴瘤患者获得的PBMC以及健康志愿者的患者以及幽门螺杆菌细胞易药的刺激。还通过包含1100个基因的cDNA微阵列分析了mRNA表达。在细胞因子和/或HSP60抗原刺激的麦芽淋巴瘤患者的PBMC中，CD40L在CD4阳性细胞上的表达增加。麦芽淋巴瘤患者的PBMCS培养物中IL-4的产生增加。但是，IFN-_γ的产生在低水平。 cDNA微阵列分析表明……更多的ED提高了HLA-DR和整联蛋白的mRNA水平。在低级别麦芽淋巴瘤的情况下，宿主因子可以通过宿主因子（例如抗原呈递和T细胞激活）增强对HSP60的适应性免疫反应，从而导致B细胞增殖，这可以在H.幽门肝炎感染中证明。在HSP60中，HSP60在HSP60中进行了HSP60在HSP60中受Immunici in Immunuction hsp60 to hsp60，我们的Immunoguty HSP60。整个HSP60和螺旋细菌HSP60的两个部分区域被表示为GST融合蛋白。指定了三种重组蛋白RHSPW，RHSP2和RHSP4-5。 RHSPW表示为整个HSP60（MET_1-MET_ <545>）。 RHSP2（GLU_ <101> -SER_ <200>）包含一个Dimaint Epipatope群集（Lys_ <159> -TNR_ <178>）。 RHSP4-5包含在T细胞表位群集中存在的域（ASP_ <396> -Gly_ <4l2>）及其在H. Pylori和人类HSP60之间共有的区域（Ser_ <356> -asp_ <392>）的区域（Ser_ <356> -asp_ <392>）。大肠杆菌的重组热能肠毒素（RLT）也被用作调整鼻免疫。将三个不同的小鼠组（BALB/C，C3H/HE，C57BL/6）每周两次注入10^8 Crj的幽门螺杆菌（SS -1），并在干净的隔离器中保存2周。这些小鼠通过在1个月内四次注入鼻粘膜（10μg/小鼠）的幽门螺杆菌裂解物或RHSP（10μg/小鼠）。最后一次免疫后，在1 MOMTN处置小鼠，然后研究组织病理学和抗体。幽门螺杆菌裂解物和RHSP的粘膜免疫抑制在BALB/C和C57BL/6小鼠中诱导严重的胃炎。在船尾组中，由RHSP4-5免疫抑制的这两组的炎症等级最高。 HSP60通过HSP60加速产生IgG和IgA的粘膜免疫抑制。这些结果表明HSP60与幽门螺杆菌诱导的胃炎密切相关。较少的