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建立 MBP-20 肽融合蛋白诱导的小鼠 IgA 肾病模型。

Establishment of a mouse IgA nephropathy model with the MBP-20-peptide fusion protein.

机构信息

Department of Pathology, Harbin Medical University, Hei Longjiang Province, People's Republic of China.

出版信息

Anat Rec (Hoboken). 2010 Oct;293(10):1729-37. doi: 10.1002/ar.21225. Epub 2010 Aug 20.

Abstract

Here, we aimed to determine whether immunoglobulin-A nephropathy (IgAN) could be induced in Balb/c mice by immunizing them with a fusion protein (MBP-20 peptide) comprising the maltose-binding protein (MBP) and a 20-amino-acid peptide derived from Staphylococcus aureus. A recombinant plasmid encoding the fusion protein was constructed and expressed in bacterial cells. The synthetic 20-peptide was used to prepare the monoclonal antibody. Balb/c mice were immunized with the MBP-20-peptide fusion protein over a 21-week course before renal histology was examined at the light and electron microscopic levels. Direct immunofluorescence staining with the anti-20-peptide monoclonal antibody was also performed using renal biopsy tissue from human IgAN patients as a comparison. IgA and IgG specific for the 20-peptide in human and mice serum were detected. The IgAN experimental mice developed a clinical and pathological profile that closely resembled that of human IgAN patients, including the induction of hematuria and numerous histopathological features. Levels of IgA and IgG specific for the 20-peptide were significantly increased in serum from the IgAN experimental mice and IgAN patients compared with control mice and non-IgAN patients. In IgAN model mice, the anti-20-peptide antibody labeled glomeruli, while the antibody strongly labeled glomeruli and weakly labeled tubular epithelial cells in renal tissue from human IgAN patients. In conclusion, immunization with an MBP-20-peptide fusion protein is able to induce clinical and pathological features closely resembling IgAN in Balb/c mice, indicating a potentially useful role for the model in the study of IgAN and related diseases.

摘要

在这里,我们旨在确定通过免疫 Balb/c 小鼠融合蛋白(MBP-20 肽)是否可以诱导免疫球蛋白 A 肾病(IgAN),该融合蛋白由麦芽糖结合蛋白(MBP)和源自金黄色葡萄球菌的 20 个氨基酸肽组成。构建了编码融合蛋白的重组质粒,并在细菌细胞中表达。使用合成的 20 肽制备单克隆抗体。在 21 周的过程中,用 MBP-20 肽融合蛋白免疫 Balb/c 小鼠,然后在光镜和电子显微镜下检查肾脏组织学。还使用来自人类 IgAN 患者的肾活检组织进行了针对 20 肽的直接免疫荧光染色,作为比较。检测了人血清和小鼠血清中针对 20 肽的 IgA 和 IgG。IgAN 实验小鼠表现出与人类 IgAN 患者非常相似的临床和病理特征,包括血尿和许多组织病理学特征的诱导。与对照小鼠和非 IgAN 患者相比,IgAN 实验小鼠和 IgAN 患者血清中针对 20 肽的 IgA 和 IgG 水平显着增加。在 IgAN 模型小鼠中,抗 20 肽抗体标记肾小球,而在来自人类 IgAN 患者的肾组织中,该抗体强烈标记肾小球并弱标记肾小管上皮细胞。总之,用 MBP-20 肽融合蛋白免疫可诱导类似于 Balb/c 小鼠 IgAN 的临床和病理特征,表明该模型在 IgAN 及相关疾病的研究中具有潜在的用途。

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