The Effect of Bruton’s Tyrosine Kinase (BTK) Inhibitor in the Eosinophilic Asthma Model of Mouse
- 주제(키워드) Asthma , Bruton's Tyrosine kinase , Eosinophil , Mast cell , T cell
- 주제(DDC) 570
- 발행기관 아주대학교 일반대학원
- 지도교수 Yoo Seob Shin
- 발행년도 2024
- 학위수여년월 2024. 2
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000033673
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Background Bruton’s Tyrosine kinase (BTK) plays a pivotal role as the key mediator in B cell receptor signaling. Recent research has revealed that it is also expressed in cells critical to asthma development, such as T cells, and eosinophils. This study aims to investigate the potential of BTK inhibitor, ibrutinib, in eosinophilic asthma mouse model. Methods BALB/c mice were sensitized with OVA via intraperitoneal injections and followed by OVA nebulizations. The mice were treated with 250ug/ml or 500ug/ml of ibrutinib before the second intraperitoneal injection and the first challenge. Two days after the last OVA challenge, airway hyperresponsiveness (AHR) was assessed with methacholine, and differential cell count in bronchoalveolar lavage fluid (BALF) was performed. The cytokines were measured in BALF, and serum OVA-specific IgE and IgG antibody levels were evaluated by ELISA. The inhibitory effect of ibrutinib was also evaluated in splenic mononuclear cells, mast cells, eosinophils, and T cells in vitro. Results Treatment with ibrutinib significantly attenuated AHR and airway inflammation, compared to the positive control. The treatment also reduced cytokine levels and suppressed OVA-specific IgE and IgG production, especially in the group treated before OVA sensitization, compared to the positive control. Additionally, ibrutinib decreased beta-hexosaminidase release from mast cells, type 2 cytokine production from mononuclear cells and T cells, and eosinophilic activation markers in vitro. Conclusion The results of this study suggest that ibrutinib treatment could exert anti- allergic effects by inactivating B cells and other BTK-expressing cells. Further studies are needed to investigate the potential therapeutic effect of ibrutinib on allergic diseases.
more목차
Ⅰ. INSTRUCTION 1
Ⅱ. MATERIALS AND METHODS 4
A. Animals 4
B. Establishment of eosinophil mouse model and ibrutinib treatment 4
C. Evaluation of airway resistance, cytokines in bronchoalveolar lavage fluid, and lung histology 5
D. Measurements of immunoglobulin levels 7
E. The assessment of cell viability by CCK-8 assay 7
F. Cell culture 8
G. Cell treatment and measurement 8
H. The isolation of mouse mononuclear cells 9
I. Detection BTK signaling pathway of western-blotting 10
J. Antibodies and reagents 11
K. Statistical analysis 12
Ⅲ. RESULTS 13
A. The effect of ibrutinib on airway hyperresponsiveness and inflammation in a mouse model of asthma 13
B. The effect of ibrutinib on the OVA specific antibody production 16
C. The ex vivo effect of ibrutinib in splenic mononuclear cells from asthmatic mice 17
D. The effect of ibrutinib in T cells 19
E. The effect of ibrutinib on IgE-sensitized mast cells 20
F. The effect of ibrutinib in eosinophils 22
G. The inhibitory effect of ibrutinib in the protein expressions of BTK signaling pathway in the mouse lung tissues 23
Ⅳ. DISCUSSION 25
Ⅴ. CONCLUSION 32
REFERENCES 33
국문요약 36
