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HMG proteins : their intra-cellular trafficking and the role in inflammatory diseases

HMG 蛋白：其细胞内运输及其在炎症性疾病中的作用

基本信息

批准号：
10470125
负责人：
OZAKI Shoichi
金额：
$ 3.71万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10470125/
关键词：
HMG proteins HMG1 HMG2 anti-HMG antibody autoimmune disease Autoimmune hepatitis epitopes intra-cellular localization 抗好中球細胞質抗体炎症性疾患 amphoterin 自己抗体

项目摘要

We found the existence of anti-HMG1/HMG2 antibodies in 89% of patients with autoimmune hepatitis. These antibodies seemed to be useful clinical markers in the disease based on the highest prevalence and the correlation with disease activity.To determine the B-cell epitopes recognized by circulating autoantibodies, we performed western blotting, in which antigens were a set of recombinant fragment peptides of HMG1 and HMG2. Antibody-positive sera from patients with autoimmune hepatitis were used. Our results indicated that the epitopes are located within a linker region between the two HMG boxes.We also investigated the mechanism of the selective staining of neutrophil by anti-HMG1/HMG2 antibodies. Affinity-purified anti-HMG1/HMG2 antibodies from patient serum reacted only with neutrophils, but not with lymphocytes, HEp-2 or HL60 cells(a promyelocytic leukemia cell line) in indirect immunofluorescence. After HL60 were treated with all-trans retinoic acid to induce granulocyte differentiation, however, these cells became reactive to the antibodies. In western blotting, the antibodies showed a distinct pattern of bands in neutrophils and differentiated HL60 cells, which was different from those in lymphocytes and untreated HL60 cells. Northern blotting revealed no significant difference in mRNA of HMG1 and HMG2 during differentiation of HL60 cells. These results indicated that the epitopes of HMG1/HGM2 molecules in neutrophils are disclosed or modified to become reactive to the antibodies during granulocyte differentiation.The linker region, which we determined to be the major epitope, is known to be necessary for the nuclear localization of HMG proteins. It seems therefore that the change of antigenicity of the region may affect the nuclear localization signal and the efficiency of transcription in mature neutrophils.

我们发现89%的自身免疫性肝炎患者存在抗HMG 1/HMG 2抗体。这些抗体似乎是有用的临床标志物在疾病的基础上的最高患病率和疾病activity.To确定的B细胞抗原表位识别的循环自身抗体，我们进行蛋白质印迹，其中抗原是一组重组片段肽HMG 1和HMG 2。使用来自自身免疫性肝炎患者的抗体阳性血清。结果表明，这些表位位于两个HMG盒之间的连接区，并初步探讨了抗HMG 1/HMG 2抗体选择性染色中性粒细胞的机制。在间接免疫荧光中，来自患者血清的亲和纯化的抗HMG 1/HMG 2抗体仅与中性粒细胞反应，而不与淋巴细胞、HEp-2或HL 60细胞（早幼粒细胞白血病细胞系）反应。然而，用全反式维甲酸处理HL 60以诱导粒细胞分化后，这些细胞变得对抗体反应。Western blotting结果显示，抗体在中性粒细胞和分化的HL 60细胞中显示出明显的条带，这与淋巴细胞和未处理的HL 60细胞中的条带不同。北方杂交显示HMG 1和HMG 2 mRNA在HL 60细胞分化过程中无明显差异。这些结果表明，中性粒细胞中HMG 1/HGM 2分子的表位在粒细胞分化过程中被暴露或修饰，从而与抗体发生反应，其中连接区是HMG蛋白核定位所必需的主要表位。因此，该区域抗原性的改变可能影响成熟中性粒细胞的核定位信号和转录效率。