Construction and validation of a targeted IL-13Rα CAR macrophage based on OX40L intracellular domain
10.19405/j. cnki.issn1000–1492.2026.07.002
- VernacularTitle:一种基于OX40L胞内域的靶向IL-13Rα CAR巨噬细胞的构建与验证
- Author:
Hui XUE
1
;
Zimeng XUE
1
;
Weile CHEN
1
;
Jiaqi QIU
1
;
Mengjuan ZHU
1
;
Yizhao CHEN
2
;
Jiajie TU
1
Author Information
1. Institute of Clinical Pharmacology， School of Pharmacy， Anhui Medical University， Key Laboratory of Anti-inflammatory and Immune Medicine （Anhui Medical University）， Ministry of Education， Hefei 230032
2. Department of Pharmacy， Third Affiliated Hospital of Anhui Medical University， Hefei 230061］
- Publication Type:Journal Article
- Keywords:
chimeric antigen receptor;
macrophages;
glioma;
phagocytosis effect;
OX40L;
immunotherapy
- From:
Acta Universitatis Medicinalis Anhui
2026;61(7):1170-1175
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo construct a chimeric antigen receptor macrophage （CAR-M） targeting interleukin-13 receptor α （IL-13Rα） and intracellular domain integration， which can double activate the signal adaptor protein OX40L of macrophages and T cells. MethodsThe CAR molecule targeting IL-13Rα （IL-13Rα-OX40L-CAR） with OX40L as the intracellular signal domain was designed and synthesized， and it was cloned into the adenovirus expression vector. After THP-1 cells were induced to differentiate into macrophages （THP-1-M） by phorbol ester， the cells were infected with recombinant adenovirus to construct THP-1 macrophages stably expressing green fluorescent protein （GFP） labeled anti-IL-13RαCAR. The expression efficiency of CAR molecules was monitored by flow cytometry. The empty vector group was used as the negative control， the CD3ζ group was used as the positive control， and the CD3ζ-OX40L group was used as the experimental group. The phagocytic function of macrophages in each group on IL-13Rα high expression glioma cells （U251） and low expression glioma cells （T98G） was detected by in vitro co-culture experiments. ResultsCAR adenovirus was efficiently transfected into THP-1-M to obtain anti-IL-13RαCAR-M. After co-cultured with glioma cells with different expression levels of IL-13Rα， the results of flow cytometry showed that there was no significant difference among the three groups for T98G cells with low expression of IL-13Rα. After co-culture with U251 cells with high expression of IL-13Rα， the CD3ζ group showed enhanced phagocytosis compared with the empty vector group （P<0.05）. Compared with the CD3ζ group， the phagocytosis ability of the CD3ζ-OX40L group was further enhanced （P<0.05）. ConclusionAnti-IL13Rα- OX40L-CAR-M is successfully constructed by constructing IL-13Rα-OX40L-CAR adenovirus and transfecting PMA-induced differentiated THP-1-M. It can engulf glioma cells with high expression of IL-13Rα by specific targeting， and shows activation of intracellular domain superior to traditional CD3ζ.
