Construction and validation of a GD2 CAR THP-1 cell line targeting the intracellular domain of DAP12
10.19405/j.cnki.issn1000-1492.2026.08.002
- VernacularTitle:一种基于DAP12胞内域的靶向GD2 CAR THP-1细胞系的构建
- Author:
Jiaqi QIU
1
;
Mengjuan ZHU
1
;
Xinlei CHEN
1
;
Lucheng ZHOU
2
;
Shuai WANG
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;
glioma;
DNAX-activating protein 12;
phagocytosis
- From:
Acta Universitatis Medicinalis Anhui
2026;61(8):1342-1347
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo obtain a construct chimeric antigen receptor macrophages （CAR-M） targeting disialoganglioside GD2 （GD2） and harboring the intracellular domain of macrophage-specific activating signaling adaptor protein DNAX-activating protein 12 （DAP12）. MethodsA CAR molecule targeting GD2 with the DAP12 intracellular domain as the signaling module ［Anti-GD2 CAR （DAP12）］ was designed， synthesized， and cloned into an adenoviral vector. This virus was used to infect PMA-differentiated THP-1 macrophages to construct green fluorescent protein （GFP）-labeled Anti-GD2 CAR THP-1 cells. CAR expression was confirmed using live-cell imaging. In vitro co-culture assays were performed to evaluate the phagocytic function of empty， Anti-GD2 CAR-M （CD3ζ）， and Anti-GD2 CAR-M （DAP12） groups against glioma cells with high （T98G） and low （U251） GD2 expression. ResultsThe CAR adenovirus efficiently transduced THP-1 macrophages， resulting in successful generation of Anti-GD2 CAR-M. Flow cytometry analysis following co-culture with glioma cells showed that compared to the empty group， the Anti-GD2 CAR-M （CD3ζ） group exhibited significantly enhanced phagocytosis （P<0.05）. Furthermore， compared to the Anti-GD2 CAR-M （CD3ζ） group， the Anti-GD2 CAR-M （DAP12） group demonstrated a further increase in phagocytic capacity specifically against GD2-high T98G cells （P<0.05）. ConclusionAnti-GD2 CAR-M （DAP12） cells are successfully established by transducing PMA-differentiated THP-1-M with recombinant Anti-GD2 CAR （DAP12） adenovirus. The engineered CAR-M can specifically target phagocytose glioma cells with high GD2 expression.
