Development of an animal model of opticospinal form of multiple sclerosis using the HLA-DP5 transgenic mice.

使用 HLA-DP5 转基因小鼠开发视脊髓形式的多发性硬化症动物模型。

• 批准号: 14370209
• 负责人: KIRA Jun-ichi
• 金额: $ 9.6万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370209/
• 关键词: multiple sclerosis; opticospinal; hsp105; HLA class II; transgenic mouse; SEREX; HSP105

1.Identification of a novel autoantigen hsp105 in patients, with multiple sclerosisTo discover novel autoantigens in Japanese patients with multiple sclerosis(MS), we immunoscreened spinal cord-derived cDNA library with sera from 8 MS patients(5-10×10^5 clones screened in each patient). One of five positive clones was heat shock protein 105(hsp105), which is expressed most abundantly in the brain. We then examined immune responses to hsp105 in MS patients and found that (1)the frequency of anti-hsp105 IgG antibody increased in MS patients than in controls, (2)hsp105 expression was enhanced in the MS lesions and EAE, (3)significant proliferation of CD4^+CD45RO^+T cell to hsp105 was noted only in MS patients, (4)in the ELISPOT assay, IL-10 response to hsp105 was significantly higher than IL-4 and IFN-gamma responses to hsp105 in both MS patients and healthy controls, however the IL-10 response was significantly lower as compared with controls. These findings suggest the immune response to hsp105 in human and that the response is attenuated in MS patients. Furthermore, we studied immunogenicity of hsp105 in mice and found that (5)immunization with either human or autologous mouse hsp105 did not induce EAE, (6)mice vaccinated with pHSP105showed marked exacerbation of EAE. In future, we are going to analyze the mechanism of exacerbation of EAE by pHSP105.2.Animal model of opticospinal form of multiple sclerosisIn Japan, susceptibility to OS-MS is associated with the HLA-DPB1*0501 allele. HLA-DP5(DPA1*02022/DPB1*0501) transgenic mice were generated by co-injection of the HLA-DPA1*02022/pDOI-6 fragment and HLA-DPB1*0501/pDOI-6 fragment into fertilized mouse eggs(C57BL/6). The resulting mice were tested for integration of the transgene by PCR analysis of tail DNA. Now, transgenic lines derived from three independent founders are tested for expression of HLA-DPA1*02022 RNA and HLA-DPB1*0501 RNA in thymus and spleen.

1。s scurosis（MS），我们从8 ms患者中进行了脊髓衍生的cDNA文库（5-10×10^5个克隆，在每个患者中都受到了最丰富的体验。 MS患者的HSP105发现，（1）MS患者的抗HSP105 IgG抗体的频率比对照组中增加，MS病变中的HSP105 Express在MS病变中得到了增强，而EAE F CD4^+CD45ro^+T细胞与HSP105不仅是在MS患者中（在ELISPOT分析中4个IL-105在MS患者和健康OLS中对HSP105的IL-10显着高，IFN-GAMMA对HSP105的反应显着降低，但是与对照组相比，IL-10响应显着降低。研究结果表明，人类的免疫RESP105在MS患者中衰减正在分析日本多发性硬化症的PHSP105的EAE的机制，对OS-MS的敏感性与HLA-DPB1*0501等位基因相关通过共同注入HLA-DPA1 *02022/pDOI-6片段和DPB1 *0501/pDOI-6片段中产生转基因。 DNA。

项目成果

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Osoekawa M 等人：“具有特应性素质的脊髓炎：对日本 79 例病例进行的全国性调查。”J Neurol Sci。

Ochi H et al.: "Proteomic analysis of human brain identifies alpha-enolase as a novel autoantigen in Hashimoto's encephalopathy."FEBS Lett. 528. 197-202 (2002)

Ochi H 等人：“人脑的蛋白质组学分析确定 α-烯醇化酶是桥本脑病的一种新型自身抗原。”FEBS Lett。

Ochi H.: "Multiple sclerosis."Karada No Kagaku. 226. 85-92 (2002)

Ochi H.：“多发性硬化症”。Karada No Kagaku。

Osoegawa M et al.: "Platelet-activating factor acetylhydrolase gene polymorphism and its activity in Japanese patients with multiple sclerosis."J Neuroimmunol. In press.

Osoekawa M 等人：“日本多发性硬化症患者的血小板激活因子乙酰水解酶基因多态性及其活性。”J Neuroimmunol。

Kira J.: "Molecular immunogenetic approach to the pathogenesis of multiple sclerosis."Clinica Neurol. 42. 1198-1200 (2002)

Kira J.：“多发性硬化症发病机制的分子免疫遗传学方法。”Clinica Neurol。