Immune-check blocking combination multiple cytokines shown curative potential in mice tumor model

Area of Science:

• Immunology
• Molecular Biology
• Cancer Therapy

Background:

• Developing effective cancer therapies requires strategies to enhance the host immune response against tumors.
• Immune checkpoint inhibitors and cytokine therapies are promising but often used separately.

Purpose of the Study:

• To construct a eukaryotic expression vector encoding PD-1v and multiple cytokines (IL-2/15, IL-12, GM-CSF) for combined immunotherapy.
• To evaluate the efficacy of this combined therapy in activating immune responses and inhibiting tumor growth in a mouse model.

Main Methods:

• Construction of a novel eukaryotic expression plasmid (pT7AMPCE) carrying genes for PD-1v, IL-2/15, IL-12, and GM-CSF.
• In vitro transfection of CT26 cells and assessment of protein expression (PD-1v, IL-12, GM-CSF) via Western blot and ELISA.
• In vivo studies involving subcutaneous inoculation of CT26 tumor cells in mice, followed by intratumoral injection of recombinant plasmids and evaluation of tumor size, survival, immune cell infiltration, and cytokine levels.

Main Results:

• Successful construction and expression of recombinant plasmids encoding PD-1v, IL-12, and GM-CSF.
• Significant inhibition of tumor growth and improved survival rates in mice treated with the combined PD-1v and cytokine therapy compared to control groups.
• Demonstrated activation of immune cells and increased immune cell infiltration (IFN-γ, TNF, IL-4, IL-2, IL-5) within tumor tissues, alongside tumor cell necrosis.

Conclusions:

• The combination of immune checkpoint blockade (PD-1v) and multiple cytokine therapy effectively activates the immune system.
• This novel therapeutic strategy demonstrates significant potential for inhibiting tumor growth and warrants further investigation for cancer treatment.