Related Experiment Video
Updated: Jun 3, 2026
Study of Dendritic Cell Development by Short Hairpin RNA-Mediated Gene Knockdown in a Hematopoietic Stem and Progenitor Cell Line In vitro
Published on: March 7, 2022
[Functional study of transcription factor Hlx modified dendritic cell line DC2.4]
Cheng-lin Zhou1, Zhao-liang Su, Jin-lian Zhu
1Zhenjiang Entry-exit Inspection and Quarantine Bureau, Zhenjiang 212008, China. zjsyzcl@126.com
Aim:
To transfect Hlx into mouse dendritic cell line DC2.4 and observe the effect of hlx on function of dendritic cells.
Methods:
The eukaryotic expression vector PIRES2-EGFP/Hlx was transfected into DC2.4 by liposomes. The transfection efficiency was identified through FACS. RT-PCR and Real-time PCR were used to test the transcription level of Hlx in DC2.4. Forty-eight hours after transfection, DC2.4 cells were studied for cytokine production, cell phenotype, phagocytosis, unilateral mixed lymphocyte reaction.
Results:
The pIRES2-EGFP/Hlx vector was transfected into DC2.4 with the transfection efficiency of up to 60%. Highly expressed Hlx in DC2.4 increased the expression of maturation makers including CD80 and CD86, and major histocompatibility complex-II. Functional assay showed that over-expression of Hlx in DC2.4 increased the interleukin-12 transcription and decreased DC endocytosis. The Hlx modified DC2.4 highly expressed IL-10 and TGF-β at the same time. Furthermore, it was shown that in a unilateral mixed lymphocyte reaction model, Hlx modified DC2.4 inhibited proliferation of lymphocytes.
Conclusion:
Transient over-expression of Hlx in DC2.4 promotes DC2.4 maturation and up-regulates IL-12, IL-10 and TGF-β expression. However, the Hlx modified DC2.4 cells functionally appear as regulatory dendritic cells.
