Development and testing of autologous in vitro models for their ability to mimic the immune system’s response toward immunotherapies
(2023) KIMM01 20231Department of Immunotechnology
Educational programmes, LTH
- Abstract
- Immunotherapy aims to enhance the immune system’s response to cancer. Different immunotherapies exist, such as cytokine therapies and immune checkpoint inhibitors. The FDA has approved immunomodulatory drugs, such as Proleukin, a recombinant IL-2 cytokine, Nivolumab, and Avelumab, which are immune checkpoint inhibitors. However, these drugs lead to side effects in patients known as immune-related adverse events such as cytokine release syndrome. Hence, there is a need to obtain models which mimic the immune system's response to examine the adverse events that could occur due to these immunotherapies. Cytokine release syndrome caused due to the overactivation of the immune system, which leads to the secretion of high amounts of cytokines... (More)
- Immunotherapy aims to enhance the immune system’s response to cancer. Different immunotherapies exist, such as cytokine therapies and immune checkpoint inhibitors. The FDA has approved immunomodulatory drugs, such as Proleukin, a recombinant IL-2 cytokine, Nivolumab, and Avelumab, which are immune checkpoint inhibitors. However, these drugs lead to side effects in patients known as immune-related adverse events such as cytokine release syndrome. Hence, there is a need to obtain models which mimic the immune system's response to examine the adverse events that could occur due to these immunotherapies. Cytokine release syndrome caused due to the overactivation of the immune system, which leads to the secretion of high amounts of cytokines such as IL-6, IL-10, and IFN-γ. In this thesis, an autologous coculture model of blood outgrowth endothelial cells and peripheral blood mononuclear cells was set up using a patient’s sample to examine the ability of various immunotherapies provoking cytokine release syndrome. The coculture model was stimulated with Proleukin, IL-2n (a novel variant of IL-2), Nivolumab, and Avelumab. A 3D model of the coculture was also obtained using plates with nanofibers to mimic a more physiologically relevant immune system environment. The stimulation with Proleukin and IL-2n was examined after 1.5 h and 24 h. Stimulation with Nivolumab and Avelumab was performed only for 24h. The supernatants were analyzed using Luminex analysis to determine the concentration of analytes presented during cytokine release syndrome. C5a was found to have the highest concentration after using Proleukin and Nivolumab.
The effects of Proleukin and IL-2n on a specific immune cell type called dendritic cells were also investigated using an autologous coculture model of monocyte-derived dendritic cells and T cells. During cancer, dendritic cells can recognize tumor-associated antigens and stimulate effector cells such as T-cells, eventually killing cancer cells. The impact of the particular immunotherapies was investigated on the activation of monocyte-derived dendritic cells in the autologous coculture compared to the monoculture of monocyte-derived dendritic cells. Additionally, the expression of the three IL-2 receptors IL-2Rα (CD25), IL-2Rβ (CD122), and IL-2Rγ (CD132) was investigated. It appears that there was an activation using Proleukin, IL-2n, and IL-2Rγ had the highest expression level. This may be because different cytokines share it. (Less) - Popular Abstract
- Immunotherapy is an example of cancer therapy which helps the immune system to eliminate cancer cells. Different drugs can be administered to cancer patients that can enhance immune cells' behavior to fight cancer. However, sometimes these drugs can stimulate the immune system too much and release massive amounts of small-sized proteins that can control the behavior of the immune cells by sending specific signals to them. This “storm” of small-sized proteins within the body creates a toxic environment that leads to side effects starting from mild ones, such as flu-like symptoms, and can develop into more severe such as organ failures and even death. To prevent it, the drugs should be tested before being administered to the patients. Hence,... (More)
- Immunotherapy is an example of cancer therapy which helps the immune system to eliminate cancer cells. Different drugs can be administered to cancer patients that can enhance immune cells' behavior to fight cancer. However, sometimes these drugs can stimulate the immune system too much and release massive amounts of small-sized proteins that can control the behavior of the immune cells by sending specific signals to them. This “storm” of small-sized proteins within the body creates a toxic environment that leads to side effects starting from mild ones, such as flu-like symptoms, and can develop into more severe such as organ failures and even death. To prevent it, the drugs should be tested before being administered to the patients. Hence, it is necessary to develop tests to predict the side effects of these drugs. This could be achieved using specific types of cells from human blood known to be responsible for releasing those small-sized proteins after immunotherapies are added to them.
In this thesis, the effect of immunotherapies on specific immune cells found in the blood was examined by determining the amount and type of essential small-sized proteins released due to the overactivation of the immune system. A more natural environment was attempted to be created using specific components to lead to a more realistic immune system response. Also, the effect of specific immunotherapies was examined on particular cells that play a crucial role in recognizing cancer cells. This project managed to get some information on the amount and type of small-sized proteins that were released after the addition of immunotherapies in specific cells derived from a cancer patient’s blood sample. In addition, this project obtained information on what is happening after the addition of immunotherapies to the cells responsible for recognizing cancer cells. To summarize, this thesis established the basis of an approach to testing immunotherapy safety and collected initial data that can be validated in future experiments. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9130711
- author
- Prastitou, Stella LU
- supervisor
- organization
- course
- KIMM01 20231
- year
- 2023
- type
- H2 - Master's Degree (Two Years)
- subject
- language
- English
- id
- 9130711
- date added to LUP
- 2023-06-27 16:07:22
- date last changed
- 2023-06-27 16:07:22
@misc{9130711,
abstract = {{Immunotherapy aims to enhance the immune system’s response to cancer. Different immunotherapies exist, such as cytokine therapies and immune checkpoint inhibitors. The FDA has approved immunomodulatory drugs, such as Proleukin, a recombinant IL-2 cytokine, Nivolumab, and Avelumab, which are immune checkpoint inhibitors. However, these drugs lead to side effects in patients known as immune-related adverse events such as cytokine release syndrome. Hence, there is a need to obtain models which mimic the immune system's response to examine the adverse events that could occur due to these immunotherapies. Cytokine release syndrome caused due to the overactivation of the immune system, which leads to the secretion of high amounts of cytokines such as IL-6, IL-10, and IFN-γ. In this thesis, an autologous coculture model of blood outgrowth endothelial cells and peripheral blood mononuclear cells was set up using a patient’s sample to examine the ability of various immunotherapies provoking cytokine release syndrome. The coculture model was stimulated with Proleukin, IL-2n (a novel variant of IL-2), Nivolumab, and Avelumab. A 3D model of the coculture was also obtained using plates with nanofibers to mimic a more physiologically relevant immune system environment. The stimulation with Proleukin and IL-2n was examined after 1.5 h and 24 h. Stimulation with Nivolumab and Avelumab was performed only for 24h. The supernatants were analyzed using Luminex analysis to determine the concentration of analytes presented during cytokine release syndrome. C5a was found to have the highest concentration after using Proleukin and Nivolumab.
The effects of Proleukin and IL-2n on a specific immune cell type called dendritic cells were also investigated using an autologous coculture model of monocyte-derived dendritic cells and T cells. During cancer, dendritic cells can recognize tumor-associated antigens and stimulate effector cells such as T-cells, eventually killing cancer cells. The impact of the particular immunotherapies was investigated on the activation of monocyte-derived dendritic cells in the autologous coculture compared to the monoculture of monocyte-derived dendritic cells. Additionally, the expression of the three IL-2 receptors IL-2Rα (CD25), IL-2Rβ (CD122), and IL-2Rγ (CD132) was investigated. It appears that there was an activation using Proleukin, IL-2n, and IL-2Rγ had the highest expression level. This may be because different cytokines share it.}},
author = {{Prastitou, Stella}},
language = {{eng}},
note = {{Student Paper}},
title = {{Development and testing of autologous in vitro models for their ability to mimic the immune system’s response toward immunotherapies}},
year = {{2023}},
}