Cold Atmospheric Plasma (CAP) is a non-equilibrium ionized gas generated under non-equilibrium discharge conditions. In this study, a stable violet gas jet was formed from a glass tube and used to treat immune cells in vitro. The reactive species produced by the CAP treatment caused a biological impact on the samples, leading to an inflammatory phenotype that enhanced antigen-presenting and effector cell function in rat peritoneal exudate macrophages and T-cells respectively. Unlike previous studies that mainly used immortalized cell lines, this study utilized primary cells isolated from mice. The biological assays performed after CAP treatment included cytokine production, surface markers, and antigen-presentation. The results showed that CAP disrupted tolerogenic pathways leading to enhanced production of pro-inflammatory cytokines while limiting the production of anti-inflammatory cytokines and the expression of inhibitory molecules such as programmed death-ligand 1 (PD-L1). This suggests that CAP represents a novel, non-toxic technology for augmenting immune cell function and enhancing antitumor responses when used as part of T-cell adoptive immunotherapies strategies or potentially in combination with other cancer immunotherapeutic approaches. To further explore its potential applications in vivo, ex vivo exposure of T-cells to CAP followed by their adoptive transfer into tumor bearing mice resulted in a strong antitumor effect. In addition to these experiments, mouse spleens or lymph nodes were harvested under sterile conditions for total splenocytes or lymphoid cells isolation followed by flow cytometry analysis using fluorochrome labeled monoclonal antibodies (mAbs; anti PD L1 , anti TIM ) for 20 minutes at 4ºC and washed twice with FACS buffer along with vitality dye 4',6 diamidino 2 phenylindole . Overall , this study demonstrates that CAP can enhance both innate and adaptive immune systems by disrupting tolerogenic pathways and promoting the production of pro inflammatory cytokines . This technology has potential applications in cancer immunotherapy as a non toxic and easy to deliver treatment to augment immune cell function and enhance antitumor responses .
Cold Atmospheric Plasma (CAP) is a special gas that can help our immune cells fight against diseases like cancer. Scientists used this gas to treat cells in a lab and found that it made the cells stronger and better at fighting diseases. They also tested the gas on mice with tumors and found that it helped their immune system fight the cancer. This new technology could be used in the future to help people with cancer.
Definitions
- Cold Atmospheric Plasma (CAP): A type of non-equilibrium ionized gas generated under special conditions.
- Immune cells: Cells in our body that help us fight off diseases.
- In vitro: A term used to describe experiments done outside of a living organism, usually in a laboratory setting.
- Cytokine: A type of protein that helps our immune system communicate and coordinate its response to diseases.
- Tolerogenic pathways: Processes in our body that prevent our immune system from attacking healthy cells or tissues.
- Antigen-presenting cell: A type of immune cell that shows other immune cells what they should attack.
- Adoptive immunotherapy strategies: Treatments where doctors take out some of a patient's own immune cells, make them stronger, and then put them back into the patient's body to help fight disease.
- Flow cytometry analysis: A technique used to count and analyze different types of cells based on their physical properties.
Cold Atmospheric Plasma (CAP): A Novel Non-Toxic Technology for Enhancing Immune Cell Function and Antitumor Responses
Cold Atmospheric Plasma (CAP) is a non-equilibrium ionized gas generated under non-equilibrium discharge conditions. It has been studied as a potential tool to enhance immune cell function and antitumor responses in cancer immunotherapy. In this study, researchers investigated the effects of CAP on primary cells isolated from mice, including rat peritoneal exudate macrophages and T-cells. The results showed that CAP disrupted tolerogenic pathways leading to enhanced production of pro-inflammatory cytokines while limiting the production of anti-inflammatory cytokines and the expression of inhibitory molecules such as programmed death ligand 1 (PD-L1). To further explore its potential applications in vivo, ex vivo exposure of T-cells to CAP followed by their adoptive transfer into tumor bearing mice resulted in a strong antitumor effect.
Formation Of Stable Violet Gas Jet
In this study, a stable violet gas jet was formed from a glass tube and used to treat immune cells in vitro. The reactive species produced by the CAP treatment caused a biological impact on the samples, leading to an inflammatory phenotype that enhanced antigen presenting and effector cell function in rat peritoneal exudate macrophages and T cells respectively. Unlike previous studies that mainly used immortalized cell lines, this study utilized primary cells isolated from mice.
Biological Assays Performed After CAP Treatment
The biological assays performed after CAP treatment included cytokine production, surface markers, and antigen presentation. Cytokine production was measured using ELISA kits specific for mouse IL6 , IFNγ , IL17A , IL10 , PD L1 . Surface marker expression was assessed using flow cytometry with fluorochrome labeled monoclonal antibodies (mAbs; anti PD L1 , anti TIM ) for 20 minutes at 4ºC followed by washing twice with FACS buffer along with vitality dye 4',6 diamidino 2 phenylindole . Antigen presentation was evaluated through intracellular staining for MHC class II molecules CD74 or HLA DR .
Results Showed Enhanced Production Of Pro Inflammatory Cytokines
The results showed that CAP disrupted tolerogenic pathways leading to enhanced production of pro inflammatory cytokines while limiting the production of anti inflammatory cytokines and the expression of inhibitory molecules such as programmed death ligand 1 (PD L1 ). This suggests that CAP represents a novel non toxic technology for augmenting immune cell function and enhancing antitumor responses when used as part of T cell adoptive immunotherapies strategies or potentially in combination with other cancer immunotherapeutic approaches.
Ex Vivo Exposure Of T Cells To Cap Followed By Adoptive Transfer Into Tumor Bearing Mice Resulted In A Strong Antitumor Effect
To further explore its potential applications in vivo, ex vivo exposure of T cells to CAP followed by their adoptive transfer into tumor bearing mice resulted in a strong antitumor effect. Mouse spleens or lymph nodes were harvested under sterile conditions for total splenocytes or lymphoid cells isolation followed by flow cytometry analysis using fluorochrome labeled monoclonal antibodies (mAbs; anti PD L1 , anti TIM ) for 20 minutes at 4ºC before being washed twice with FACS buffer along with vitality dye 4',6 diamidino 2 phenylindole .
Conclusion
Overall , this study demonstrates that Cold Atmospheric Plasma can enhance both innate and adaptive immune systems by disrupting tolerogenic pathways and promoting the production of pro inflammatory cytokines . This technology has potential applications in cancer immunotherapy as an easy to deliver treatment which could be used alone or combined with other therapies due its non toxic nature which augments immune cell function thereby enhancing antitumor responses .
Cold atmospheric plasma (CAP) is a near room temperature ionized gas, generated under non-equilibrium discharge conditions. Here we show that a short exposure of rat peritoneal exudate macrophages and T-cells to CAP in vitro, triggered an inflammatory phenotype leading to better antigen-presenting and effector cell function respectively. Different from previous studies mainly using immortalized cell lines, both macrophage and T-cells in this study were primary cells isolated from mice. Furthermore, ex-vivo exposure of T-cells to CAP, followed by their adoptive transfer into tumor-bearing mice resulted in a strong antitumor effect in vivo. Mechanistically, CAP seems to disrupt tolerogenic pathways leading to enhanced production of pro-inflammatory cytokines while limiting the production of anti-inflammatory cytokines and the expression of inhibitory molecules such as programmed death-ligand 1 (PD-L1). CAP represents therefore a novel, non-toxic and easy to deliver technology to augment the function of immune cells and enhance antitumor responses when used as a component of T-cell adoptive immunotherapies strategies or, potentially in combination with other cancer immunotherapeutic approaches.
