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.在患者中识别一种新型的自身抗原HSP105，多发性硬化症发现了日本多发性硬化症患者（MS）患者的新型自身抗原，我们通过8 ms患者的血清（5-10×10^5个患者筛查的5-10×10^5个clones）免疫脊髓衍生的cDNA库。五个阳性克隆之一是热休克蛋白105（HSP105），它在大脑中表现得最多。然后，我们检查了MS患者对HSP105对HSP105的免疫反应，发现（1）MS患者的抗HSP105 IgG抗体的频率比对照组增加了，（2）HSP105在MS病变中增强了HSP105的表达，（3）CD45ro^+T细胞对HSP105的响应显着增殖，仅ELIS（4）（4）（4）（4）。 HSP105显着高于IL-4，MS患者和健康对照组中对HSP105的IFN-GAMMA响应，但是与对照组相比，IL-10响应明显低。这些发现表明，人类对HSP105的免疫响应，MS患者的反应减弱。此外，我们研究了小鼠中HSP105的免疫原性，发现（5）用人或自体小鼠HSP105免疫不会诱导EAE，（6）用PHSP105Showshow的小鼠用PHSP105SHOWSHOW SHOW SHOW SHAW SHOW SARKED SARKEDE EAE的病变。将来，我们将分析日本多发性硬化症的视脊髓形式的Animimal模型加剧EAE的机制，对OS-MS的敏感性与HLA-DPB1*0501等位基因有关。 HLA-DP5（DPA1*02022/DPB1*0501）通过共注射HLA-DPA1*02022/PDOI-6片段以及HLA-DPB1*0501/PDOI-6片段来产生转基因小鼠。通过PCR分析尾巴DNA测试了所得小鼠的转化整合。现在，对三个独立创始人得出的转基因线进行了测试，以表达HLA-DPA1*02022 RNA和HLA-DPB1*0501 RNA在胸腺和Spleen中。

项目成果

Osoegawa M et al.: "Myelitis with atopic diathesis : a nationwide survey of 79 cases in Japan."J Neurol Sci. (In press).

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。

吉良潤一: "多発性硬化症の臨床疫学-環境原因と遺伝原因"日本臨床. 61. 1300-1310 (2003)

Junichi Kira：“多发性硬化症的临床流行病学 - 环境原因和遗传原因”日本临床 61. 1300-1310 (2003)

吉良潤一: "多発性硬化症の臨床疫学-環境要因と遺伝要因"日本臨床. (In press).

Junichi Kira：“多发性硬化症的临床流行病学 - 环境和遗传因素”日本临床（正在出版）。

吉良潤一: "日本の多発性硬化症"Current Insights in Neurological Science -神経内科・脳神経外科・精神科の最新トピックス. (In press).

Junichi Kira：“日本的多发性硬化症”神经科学的最新见解 - 神经病学、神经外科和精神病学的最新主题（正在出版）。

越智博文, 吉良潤一: "Annual Review 神経 2003"柳澤信夫、篠原幸人、岩田誠、清水輝夫、寺本明 編 (中外医学社). 360 (2003)

Hirofumi Ochi、Junichi Kira：“Annual Review Neurology 2003”，由 Nobuo Yanagisawa、Yukito Shinohara、Makoto Iwata、Teruo Shimizu 和 Akira Teramoto 编辑（Chugai Igakusha）（2003 年）。