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Archive > Volume 20 Issue 5 > 2013,20(5):559-564. DOI:10.3872/j.issn.1007-385X.2013.05.009 Prev Next
Basic Research
Screening of Jurkat cells expressing FLT1-CAR and their chemotaxis to VEGF
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Abstract:
Objective:To construct Jurkat cells expressing chimeric antigen receptor (CAR) of vascular endothelial growth factor receptor 1 (VEGFR1/FLT1) using lentivirus and to investigate their chemotaxis to VEGF. Methods: The CAR targeting VEGFR1 was synthetized, and the recombinant lentiviral vector named LV-gn-FLT1-CAR was constructed and infected into Jurkat cells. Stable infected cell stains were obtained through G418 screening. The expressions of FLT1-CAR mRNA and protein in the stable infected cell stains were detected by PCR and flow cytometry. The chemotaxis of selected cell strains toward VEGF was detected by Transwell assay. Results: Recombinant lentivirus LV-gn-FLT1-CAR was successfully constructed. FLT1-CAR was successfully integrated into Jurkat cells and FLT1-CAR protein was stably expressed. The selected cell strains, Jurkat-gn-FLT1CAR-1 and Jurkat-gn-FLT1CAR-2, showed obvious chemotaxis to VEGF. In a VEGF concentration of 100 ng/ml, Jurkat-gn-FLT1-CAR-1 cell chemotactic number was (62±8)，which was significantly higher than that of the control group Jurkat cells (18±5) (P<0.01). Conclusion: Jurkat cell strains stably express FLT1-CAR are successfully obtained, which have a distinct chemotaxis to VEGF.
Keywords:
vascular endothelial growth factor ; chemokine (C-X-C motif) receptor 3（CXCR3） ; chimeric antigen receptor ; Jurkat cell ; chemotaxis
Project Supported:
Project supported by the Major Special Foundation from Ministry of Science and Technology of China (No. 2012ZX0002014-005), and the Genearl Program of the National Natural Science Foundation of China (No.810850)
