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Studies on human complement 4 binding protein

人补体4结合蛋白的研究

基本信息

批准号：
03670325
负责人：
OKAMURA Seiichi
金额：
$ 1.34万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670325/
关键词：
Complement regulatory protein Genomic organization Complement 4 binding-protein Acute phase reactive protein Monoclonal antibody Rheumatoid arthritis Cytokine モノクロ-ナル抗体

项目摘要

1. Purification of human C4-binding protein (C4BP) and development of antibody against the purified C4BP Human C4BPalpha was purified from 1.5L of plasma by multiple steps including BaC12 precipitation, Intralipid absorption, ultracentrifugation, ethanol/ethel extraction, column chromatographies (DE52,S400 and S300). Antibodies against this protein were raised in rabbits and mice. Monoclonal antibody (IgG) was also developed. 2. Human genomic C4BP gene Human genomic C4BPalpha gene was analyzed. This gene consists of 12 exons and spans about 40 kb. Each of the SCRs is encoded by a single exon, except for the second SCR (SCR II), which is encoded by two separate exons. The 5' flanking region was sequenced up to 380 bases upstream from the putative transcription initiation site. Several possible binding sites for transcription factors were identified. 3. Gene expression of the C4BPalpha The gene expression of the C4BP alpha in normal liver cell and hepatoma lines was demonstrated. The constitutive gene expression of C4BPalpha by a hepatoma line, HepG2, was significantly augmented by treatment with monocyte conditioned medium, 12-O-tetradecanoylphorbol-13-acetate,interleukin-6 and tumor necrosis factor but not by a calcium ionophore or interleukin-1beta. 4. Serum levels of C4BP Serum levels of C4BP were measured in patients with various autoimmune disease. Significantly high level of C4BP was observed in patients with active or LAC-positive SLE,Behcet disease, aortitis, PSS,RA and MCTD. The level of C4BP was correlated with CRP, C3 and CH50.

1.人C4结合蛋白(C4BP)的纯化及抗C4BP抗体的研制经BaC12沉淀、磷脂吸附、超速离心、乙醇/乙醇提取、柱层析(DE52、S400、S300)等步骤从1.5L血浆中纯化人C4BPalpha。在兔子和小鼠中产生了针对这种蛋白的抗体。还研制了单抗(Ig G)。2.分析了人类基因组C4BP基因。该基因由12个外显子组成，全长约40kb。除了第二个SCR(SCR II)外，每个SCR都由一个外显子编码，它由两个单独的外显子编码。5‘侧翼区位于转录起始点上游380个碱基。确定了几个可能的转录因子结合位点。3.C4BPalpha的基因表达C4BPα在正常肝细胞和肝癌细胞中的基因表达。经单核细胞条件培养液、12-O-十四酰佛波醇-13-乙酸酯、白介素6和肿瘤坏死因子处理后，肝癌细胞株HepG2的C4BPalpha组成基因表达显著增强，而钙离子载体和白介素1β不能显著增强C4BPalpha的表达。4.检测各种自身免疫性疾病患者的血清C4BP水平。活动期或LAC阳性的SLE、白塞病、大动脉炎、PSS、RA和MCTD患者的C4BP水平显著升高。C4BP水平与C反应蛋白、补体C3、CH50呈正相关。