Abstract
High Mobility Group Chromosomal Protein N2 (HMGN2) can recognize tumor cells and enhance the anti-tumor effect of immune cells. This study aimed to establish a lentiviral vector of recombinant HMGN2 gene, establish recombinant T cells (HMGN2-T cells), and observe their anti-tumor effects. Total RNA was isolated from peripheral blood mononuclear cells. HMGN2, cluster of differentiation (CD) 8 A, CD28, CD137, and CD3ζ genes were amplified and connected. Jurkat cells were transfected with the recombinant lentivirus vector. The viability, apoptosis, and cell cycle of HMGN2-T cells were detected using Cell Counting Kit-8 assay and flow cytometry. The co-culture was performed by adding HMGN2-T cells to tumor cells with different effect-to-target (E:T) ratios. The cytotoxic activity was measured by lactate dehydrogenase (LDH) releasing assay. The sequences of HMGN2, CD8A, CD28, CD137, and CD3ζ gene plasmids were confirmed using gene sequencing. After the lentiviral transfection for 72 h, green fluorescence cells (HMGN2-T cells) could be seen. Cell viability and apoptosis were increased in HMGN2-T cells. The cytokine levels of interleukin 2 (IL-2) and tumor necrosis factor α (TNF-α) increased in cell supernatants of HMGN2-T cells. The percentage of G0/G1 phase cells was lower, the rate of S phase cells was higher in HMGN2-T cells than control cells. The co-culture of HMGN2-T cells and tumor cells could promote the cytokines’ release. The LDH level was increased with the elevation of E:T ratios. In conclusion, the HMGN2-T cells were well-established and have the effect of secreting cytokines and killing tumor cells.
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Data Availability
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- CIK:
-
Cytokine-induced killer
- NK:
-
Natural killer
- CD:
-
Cluster of differentiation
- LAK:
-
Lymphokine-activated killer
- TCR:
-
T cell receptor
- CAR:
-
Chimeric antigen receptor
- HMGN2:
-
High Mobility Group Chromosal Protein N2
- PBMC:
-
Peripheral blood mononuclear cell
- EDTA:
-
Ethylene Diamine Tetraacetic Acid
- PBS:
-
Phosphate buffer saline
- CCK-8:
-
Cell Counting Kit-8
- ATCC:
-
American Type Culture Collection
- FBS:
-
Fetal bovine serum
- IL-2:
-
Interleukin 2
- IFN-γ:
-
Interferon γ
- TNF-α:
-
Tumor necrosis factor α
- ELISA:
-
Enzyme-Linked Immunosorbent Assay
- LDH:
-
Lactate dehydrogenase
- SD:
-
Standard deviation
- ANOVA:
-
Analysis of variance
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Li, H., Wu, X., Bu, D. et al. Recombinant jurkat cells (HMGN2-T cells) secrete cytokines and inhibit the growth of tumor cells. J Mol Histol 53, 741–751 (2022). https://doi.org/10.1007/s10735-022-10084-8
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DOI: https://doi.org/10.1007/s10735-022-10084-8