Roles of HSP60 in Helicobacter pylori infection

HSP60 在幽门螺杆菌感染中的作用

基本信息

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

项目摘要

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

我们先前报道说，休克蛋白60KDA（HSP60）是粘膜相关淋巴样淋巴瘤（MALT）淋巴瘤的重要抗原。用HSP60分析了患者的PBMC）。还通过细胞因子和 /或Hsp60抗原刺激的麦芽淋巴瘤患者的PBMC中的宽松分析。在低度麦芽淋巴瘤，适应性免疫反应的病例中，HLA-DR和整联蛋白的mRNA水平升高，可以通过宿主因子增强，导致B细胞增殖，这可以在幽门螺杆菌感染中进行。幽门螺杆菌的整个HSP60在T -epiLope群集（ASP_ <396> -gly_ <4l2>）中，其在H. Pylori和Human HSP60之间共有的区域（Ser_ <356> -asp_ <392>）。大肠杆菌的RLT也采用了三种不同的小鼠组（BALB/C，C3H/HE，C57BL/6）H.幽门杆菌（SS -1），每周两次几周。 HSP60的船尾最高的IgG和IgA产生。这些结果表明HSP60与H. RI诱导的胃炎密切相关