[Inhibitory Effect of Interference with PD-L1 Expression on B-cell lymphoma in Mice]
Qing-Qing Shi1, Miao Zhu1, Xin Sun1
1Laboratory of Hematonosis, Northern Jiangsu People's Hospital, Yangzhou Institute of Hematology, Yangzhou 225001, Jiangsu Province, China.
Objective:
To investigate the potential inhibitory effect of interference with PD-L1 on B cell lymphoma in mice.
Methods:
Three shRNA vectors for mouse CD274 (PD-L1) were constructed and transiently transfected into 293T cells. RT-qPCR was used to validate the interference efficiency of CD274. The shRNA vector that interfere efficiently with CD274 expression was packaged by using lentivirus packaging system to generate shRNA lentivirus, and then transfected into A20 lymphoma cell line. The methyl thiazol terazolium (MTT) assay was used to detect proliferation after 48 h culture of CD274-sh A20 cells. Meanwhile, BALB/c mice were hypodermically infected with CD274-sh A20 cells. Infected mice were observed daily and assessed to visualize tumor by in vivo fluorescence imaging.
Results:
The proliferation rate of CD274-sh A20 cells in vitro was significantly lower than that of A20 cells (P<0.05). The tumor size detected by in vivo fluorescence imaging showed a significant reduce in tumor bearing mice with CD274-sh compared with other tumor bearing mice. And the weight and size of tumor in CD274-sh group were also significantly reduced compared with other group (P<0.05). Moreover, the survival time of tumor bearing mice in CD274-sh group was longer than that of the PD-L1 high expression group.
Conclusion:
PD-L1 plays an important role in the incidence and the progression of lymphoma, and the shRNA-based PD-L1 knockdown can inhibit cell proliferation of A20 cells and partly suppress tumor growth.
Insights
Targeting programmed death-ligand 1 (PD-L1) with short hairpin RNA (shRNA) in mice significantly inhibited B cell lymphoma growth. This PD-L1 interference reduced tumor proliferation and extended survival, indicating its therapeutic potential.
Area of Science:
- Immunology
- Oncology
- Molecular Biology
Background:
- Programmed death-ligand 1 (PD-L1) is implicated in immune evasion by various cancers.
- Understanding PD-L1's role in B cell lymphoma is crucial for developing targeted therapies.
Purpose of the Study:
- To investigate the inhibitory effect of PD-L1 interference on B cell lymphoma progression in a murine model.
- To assess the impact of CD274 (PD-L1) knockdown on lymphoma cell proliferation and tumor growth.
Main Methods:
- Constructed and validated shRNA vectors targeting mouse CD274 (PD-L1) for efficient knockdown.
- Transfected A20 lymphoma cells with lentivirus-packaged CD274-shRNA.
- Assessed cell proliferation using MTT assay and tumor growth in vivo using fluorescence imaging in BALB/c mice.
Main Results:
- CD274-shRNA significantly reduced A20 lymphoma cell proliferation in vitro.
- In vivo, CD274-shRNA treatment led to significant reductions in tumor size, weight, and enhanced survival in tumor-bearing mice.
- Tumor growth was significantly suppressed in mice treated with CD274-shRNA compared to controls.
Conclusions:
- PD-L1 plays a critical role in the development and progression of lymphoma.
- shRNA-mediated PD-L1 knockdown effectively inhibits lymphoma cell proliferation and suppresses tumor growth.
- Targeting PD-L1 represents a promising strategy for B cell lymphoma treatment.
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