IgG78-DM1 inhibits pulmonary fibrosis by targeting and killing CD248-positive myofibroblasts in mice.
- Author:
Jingyu WANG
1
;
Ming WEI
2
;
Zhengxuan LI
3
;
Yike ZHOU
3
;
Donghui HAN
4
Author Information
1. 4th Cadet Regiment, Basic Medical Science Academy, Air Force Medical University, Xi'an 710032, China.
2. Department of Urology, 989th Hospital of Joint Logistic Support Force of Chinese People's Liberation Army, Luoyang 471032, China.
3. Department of Urology, First Affiliated Hospital, Air Force Medical University, Xi'an 710032, China.
4. Department of Urology, First Affiliated Hospital, Air Force Medical University, Xi'an 710032, China. *Corresponding author, E-mail: 906812205@qq.com.
- Publication Type:Journal Article
- MeSH:
Humans;
Animals;
Mice;
Male;
Mice, Inbred C57BL;
Pulmonary Fibrosis/drug therapy*;
Myofibroblasts;
Antibodies;
Bleomycin;
Antigens, Neoplasm;
Antigens, CD
- From:
Chinese Journal of Cellular and Molecular Immunology
2023;39(9):769-776
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the therapeutic effect of targeting and killing CD248-positive myofibroblasts on bleomycin-induced pulmonary fibrosis in mice. Methods IgG78-DM1, an antibody-maytansine 1 (DM1) conjugate targeting CD248, was prepared. The drug conjugation efficiency was measured and calculated by UV spectrophotometer, and the identification of IgG78-DM1 was performed through SDS-PAGE and Western blot analysis. In vitro, the binding activity of IgG78-DM1 on CD248-positive myofibroblasts was detected by flow cytometry and the cytotoxicity of IgG78-DM1 to CD248-positive myofibroblasts was evaluated by CCK-8 assay. In vivo, C57BL/6 male mice were randomly divided into control group, idiopathic pulmonary fibrosis group, human IgG-DM1 (hIgG-DM1) control group, and IgG78-DM1 treatment group. Then, the mouse models with pulmonary fibrosis induced by bleomycin were constructed. Two weeks later, the animal models were intravenously injected with IgG78-DM1. After the treatment of two weeks, lung tissues were collected for Masson staining and Sirius Red staining to evaluate the degree of pulmonary fibrosis. Real-time fluorescence quantitative PCR was used to measure the expression levels of CD248, as well as markers of fibroblastic activation including alpha-smooth muscle actin (α-SMA) and type I collagen alpha 1 (COL1A1). The safety of IgG78-DM1 was preliminarily assessed by conducting liver and kidney function tests. Results IgG78-DM1 was successfully prepared, and its drug conjugation ratio was 3.2. The antibody structure remained stable after conjugation, allowing effective binding and cytotoxicity against CD248-positive myofibroblasts. After treatment with IgG78-DM1, the degree of pulmonary fibrosis in mice significantly reduced, accompanied by the decrease of the expression of CD248, α-SMA, and COL1A1. The liver and kidney function of the mice remained at normal levels compared to the normal control group. Conclusion IgG78-DM1 effectively inhibits pulmonary fibrosis in mice by targeting and killing CD248-positive myofibroblasts. The safety of this strategy is preliminarily assessed.
