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Tailoring In Vivo Cytotoxicity Assays to Study Immunodominance in Tumor-specific CD8+ T Cell Responses
Published on: May 6, 2019
Investigation on the effects of soluble programmed death-1 (sPD-1) enhancing anti-tumor immune response
Ye Yuan1, Yufei He, Xiaohong Wang
1Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Abstract:
By using semi-quantitative RT-PCR method, it was found that PD-L1 mRNA but not PD-L2 mRNA was expressed in H22 hepatoma cells and both PD-L1 and PD-L2 mRNAs were expressed in tumor tissues of tumor-bearing mice and upregulated as compared with muscle tissues in normal mice and H22 hepatoma cells. PD-L1 and PD-L2 were also expressed on the surface of the activated T cells. The soluble recombinant sPD-1 expressed from the constructed eukaryotic expression vector could enhance the lysis of tumor cells by lymphocytes stimulated specifically with antigen. The expresssion of sPD-1 by local gene therapy on the inoculation site of H22 hepatoma cells could inhibit the growth of tumor. The results of this study indicate that expression of soluble receptor of negative costimulatory molecules could reduce the inhibitory effect on T cells in tumor microenvironment and enhance the cytotoxicity of T cells on tumor cells. This possibly provides a new method of improving efficacy of tumor gene therapy.
Insights
This study shows that soluble PD-1 (sPD-1) gene therapy can inhibit tumor growth by enhancing T cell activity against cancer cells. This approach may improve tumor gene therapy effectiveness.
Area of Science:
- Immunology
- Molecular Biology
- Oncology
Background:
- Programmed death-ligand 1 (PD-L1) and Programmed death-ligand 2 (PD-L2) are key negative costimulatory molecules involved in immune regulation.
- Understanding their expression in the tumor microenvironment is crucial for developing effective cancer immunotherapies.
Purpose of the Study:
- To investigate the expression of PD-L1 and PD-L2 in H22 hepatoma cells and tumor tissues.
- To evaluate the therapeutic potential of soluble PD-1 (sPD-1) gene therapy in inhibiting tumor growth.
Main Methods:
- Semi-quantitative RT-PCR was used to analyze PD-L1 and PD-L2 mRNA expression.
- Eukaryotic expression vectors were constructed to produce soluble recombinant sPD-1.
- Local gene therapy with sPD-1 was applied to H22 hepatoma cell inoculation sites in mice.
Main Results:
- PD-L1 mRNA was detected in H22 hepatoma cells, while both PD-L1 and PD-L2 mRNAs were upregulated in tumor tissues compared to normal tissues.
- PD-L1 and PD-L2 were expressed on activated T cells.
- Recombinant sPD-1 enhanced lymphocyte-mediated tumor cell lysis, and local sPD-1 gene therapy inhibited tumor growth.
Conclusions:
- Soluble receptor expression, like sPD-1, can mitigate T cell inhibition within the tumor microenvironment.
- This strategy enhances T cell cytotoxicity against tumor cells, offering a potential new avenue for improving tumor gene therapy efficacy.
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