一种新型的针对结缔组织生长因子的单链 Fv 抗体可减轻博来霉素诱导的小鼠肺纤维化。

A novel single-chain-Fv antibody against connective tissue growth factor attenuates bleomycin-induced pulmonary fibrosis in mice.

机构信息

Respiratory Department, Institute of Biotechnology and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China.

出版信息

Respirology. 2011 Apr;16(3):500-7. doi: 10.1111/j.1440-1843.2011.01938.x.

DOI:10.1111/j.1440-1843.2011.01938.x

PMID:21265950

Abstract

BACKGROUND AND OBJECTIVE

Connective tissue growth factor (CTGF) has been identified as playing critical roles in fibrosis and is a promising therapeutic target. In a previous study, we used a phage display library to develop a humanized single-chain variable fragment antibody (scFv) against CTGF. In the present study, the protective effect of anti-CTGF scFv against bleomycin (BL)-induced pulmonary fibrosis was investigated in mice.

METHODS

The expression of α-smooth muscle actin in human embryonic lung fibroblast (HELF) cells was analysed by western blotting. A mouse model of pulmonary fibrosis was established by tracheal injection of BL (5 mg/kg). Mice received anti-CTGF scFv (4 mg/kg, three times a week) by i.v. injection. The effects of anti-CTGF scFv were evaluated by leukocyte counts in BAL fluid, hydroxyproline measurements in lung tissue and pathological examination.

RESULTS

α-Smooth muscle actin expression was decreased in HELF cells treated with anti-CTGF scFv. Anti-CTGF scFv significantly reduced the numbers of inflammatory leukocytes (total and differential count) in BAL fluid, as well as the hydroxyproline content of lung tissue. The severity of alveolitis and fibrosis in the mouse model was markedly attenuated by treatment with anti-CTGF scFv.

CONCLUSIONS

Anti-CTGF scFv may potentially be developed as a useful inhibitor of pulmonary fibrosis.

摘要

背景与目的

结缔组织生长因子（CTGF）在纤维化中起着关键作用，是一种很有前途的治疗靶点。在之前的研究中，我们使用噬菌体展示文库开发了针对 CTGF 的人源化单链可变片段抗体（scFv）。在本研究中，我们研究了抗 CTGF scFv 对博来霉素（BL）诱导的肺纤维化的保护作用。

方法

通过 Western blot 分析人胚肺成纤维细胞（HELF）中α-平滑肌肌动蛋白的表达。通过气管内注射 BL（5mg/kg）建立肺纤维化小鼠模型。通过静脉注射给予抗 CTGF scFv（4mg/kg，每周 3 次）。通过 BAL 液中的白细胞计数、肺组织羟脯氨酸测量和组织病理学检查评估抗 CTGF scFv 的作用。