Construction of a MYD88-constitutively CAR THP-1M cell line targeting GD2
10.19405/j.cnki.issn1000-1492.2026.08.014
- VernacularTitle:一种基于MYD88组成型靶向GD2 CAR THP-1M细胞系的构建
- Author:
Mengjuan ZHU
1
;
Jiaqi QIU
1
;
Xinlei CHEN
1
;
Shuai WANG
2
;
Lucheng ZHOU
2
;
Yizhao CHEN
3
;
Ning LIN
2
;
Jiajie TU
1
Author Information
1. Institute of Clinical Pharmacology， School of Pharmacy， Anhui Medical University， Key Lab of Anti-inflammatory and Immune Medicine （Anhui Medical University）， Ministry of Education， Hefei 230032
2. Department of Neurosurgery， The First People's Hospital of Chuzhou， The Affiliated Chuzhou Hospital of Anhui Medical University， Chuzhou 239001
3. Department of Pharmacy， The Third Affiliated Hospital of Anhui Medical University， Hefei 230061］
- Publication Type:Journal Article
- Keywords:
chimeric antigen receptor macrophage;
myeloid differentiation factor 88;
ganglioside;
inflammatory polarization;
glioma;
phagocytosis
- From:
Acta Universitatis Medicinalis Anhui
2026;61(8):1439-1445
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo obtain stably transfected chimeric antigen receptor macrophages （CAR-M） targeting disialosylganglioside （GD2） based on the constitutive expression of myeloid differentiation factor 88 （MYD88）. MethodsAn Anti-GD2 CAR adenoviral vector incorporating the CD3ζ （cluster of differentiation 3 zeta chain）-MYD88 fusion construct was designed and constructed. THP-1-derived macrophages （THP-1M）， induced by phorbol 12-myristate 13-acetate （PMA） differentiation， were infected with this adenovirus to generate Anti-GD2 CAR THP-1 cells bearing a green fluorescent protein （GFP） tag. CAR expression was monitored using a live-cell imaging system. The phagocytic function of Empty CAR-M， Anti-GD2 CAR-M（CD3ζ）， and Anti-GD2 CAR-M（CD3ζ-MYD88） groups against glioma cells was evaluated in vitro through co-culture experiments， and their polarization status was also analyzed. ResultsCAR adenovirus successfully transfected THP-1 to obtain Anti-GD2 CAR-M. After co-culture with glioma cells， flow cytometry results showed that compared with the Empty CAR-M group and the Anti-GD2 CAR-M（CD3ζ） group， the Anti-GD2 CAR-M（CD3ζ-MYD88） group exhibited enhanced phagocytosis against T98G cells with high GD2 expression （P<0.05） and also showed upregulation of CD86 （P<0.05）. Compared with the Empty CAR-M group and the Anti-GD2 CAR-M（CD3ζ） group， the Anti-GD2 CAR-M（CD3ζ-MYD88）CAR-M group exhibited enhanced phagocytosis against T98G cells with high GD2 expression （P<0.05） and also showed upregulation of CD86 （P<0.05）， and the expression of inflammatory cytokines interleukin （IL）-6 and IL-12 was increased in this group （P<0.05）. ConclusionThis study successfully constructed Anti-GD2 CAR-M cells and verified their phagocytic activity， demonstrating that constitutive expression of MYD88 can promote inflammatory polarization of CAR-M. This provides a potential structural modification approach for the development of pro-inflammatory CAR-M.
