Establishment of an animal model for P-ANCA positive crescentic glomerulonephritis

Primary crescentiic Glomerulonepritis是快速渐进的和不容易渐进的。治疗主要取决于葡萄糖皮质激素激素和免疫抑制剂。动物模型扮演关键角色，以建立一个新的策略，用于治疗可感染性疾病。在这一研究中，我试图建立严重的结合免疫缺陷（SCID），老鼠携带经核蛋白反中性细胞浆抗体（P-ANCA）和crescentic glomerulonepritis（GN）。P-ANCAGN和HLA II类基因型之间的联系;利用PCR-RFLP和PCR-SSO;我确定了34名日本患者的HLA II类基因型。I found significant of P-ANCAGN with HLA-DRB1-D1* D10803 （Odd's ratio 2.5 p<0.01） and with HLA-DQB1-D1* D10601 （Odd's ratio 26 p<0.01）， DRB1-D10803 and DQB1-D10601 are in linkage disequiribrium in Japanese population. 2.在病人和老鼠中检测人类P-ANCA。固体相酶连接的免疫吸附测定曾用于检测人类P-ANCA。在短时间内，普尔 ... More 改良的髓过氧化物酶（MPO）被非流动化在塑料微滴定板上，样品血清被应用，良好地洗涤，碱性磷酸酶标签山羊被添加抗人类IgG或IgA抗体，PNPP被添加为基础。标题确定了我的实验室与SRL实验室中的那些密切相关。MPO特定的T细胞增殖.外围淋巴细胞在7天内被纯化的MPO，并且该增殖已被证实为先前的tritiated thymidine。没有T细胞的特定反应是演示的。SCID小鼠生产P-ANCA.外围淋巴细胞从P-ANCAGN转移到8周龄SCID小鼠。人体重量、系统性和diastolic血液压力、尿蛋白质、人类IgG和IgM以及人类P-ANCA每两周监测一次。老鼠在接下来的12周里被杀害了。人类IgG和IgM在含有人类淋巴细胞的老鼠中检测到，但只在含有患者淋巴细胞的P-ANCA中检测到。老鼠死在终点前，而且所有的老鼠都是转移的病人淋巴细胞。没有显示出老鼠蛋白质或高血压建议P-ANCAGN，病理学检查是在调查下进行的。Less（低）

Primary crescentic glomerulonephritis is rapidly progressive and poorly prognostic. The treatment depends mainly on glucocorticoid hormone and imuunosuppressants. Animal models play pivotal roles to establish new strategy for therapy of intractable of intractable diseases. In this study, I tried to raise severe combined immunodeficiency (SCID) mice carrying perinuclear anti-neutrophil cytoplasmic antibody (P-ANCA) and crescentic glomerulonephritis (GN).1. Association between P-ANCAGN and HLA class II genotypes.Utilizing PCR-RFLP and PCR-SSO, I determined HLA class II genotypes of 34 Japanese patients with P-ANCAGN. I found significant of P-ANCAGN with HLA-DRB1ィイD1*ィエD10803 (Odd's ratio 2.5 p<0.01) and with HLA-DQB1ィイD1*ィエD10601 (Odd's ratio 26 p<0.01), DRB1ィイD1*ィエD10803 and DQB1ィイD1*ィエD10601 are in linkage disequiribrium in Japanese population.2. Detection of human P-ANCA in patients and mice.Solid phase enzyme linked immunoadsorbent assay was used to detect human P-ANCA. In brief, pur … More ified myeloperoxidase (MPO) was immobilized on a plastic microtiter plate, sample serum was applied, wells were washed extensively, alkaline phosphatase labeled goat anti-human IgG or IgA antibody was added, and PNPP was added as substrate. The titers determined at my lab were well correlated with those from SRL laboratory.3. MPO specific T cell proliferation.Peripheral lymphocytes were cocultured with purified MPO for 7 days, and the proliferation was assayed by uptake of tritiated thymidine. No T cell specific response was demonstrated.4. SCID mouse producing P-ANCA.Peripheral lymphocytes from P-ANCAGN were transferred to 8-week-age SCID mice. The body weight, systolic and diastolic blood pressure, urine proteinuria, human IgG and IgM, and human P-ANCA were monitored every two weeks. The mice were sacrificed at 12 weeks of follow-up. Human IgG and IgM were detected in mice with human lymphocytes, but P-ANCA only in those with patient lymphocytes. The mice died before end point, and all were mice transferred patient lymphocyte. No mice showed proteinuria or hypertension suggesting P-ANCAGN, Pathological examination is under investigation. Less