Effect of bronchial epithelial mediators on T-cell differentiation and activation
(2024) MOBN03 20232Degree Projects in Molecular Biology
- Abstract
- Introduction: Asthma, a heterogenous respiratory disease that affects 300 million people, is characterized by coughing, chest tightness, airway inflammation, and shortening of breath. Asthma exacerbations are commonly triggered by viral infection, such as rhinovirus infection. Respiratory viral infections primarily target the epithelium resulting in the release of alarmins and proinflammatory cytokines. This leads to the recruitment of immune cells, such as T-cells, and orchestration of inflammation. The interaction between the epithelium and T-cells is relatively underexplored in asthma, thus, this study aims to create an in vitro model to investigate the effect of mediators released from viral-infected healthy and asthma bronchial... (More)
- Introduction: Asthma, a heterogenous respiratory disease that affects 300 million people, is characterized by coughing, chest tightness, airway inflammation, and shortening of breath. Asthma exacerbations are commonly triggered by viral infection, such as rhinovirus infection. Respiratory viral infections primarily target the epithelium resulting in the release of alarmins and proinflammatory cytokines. This leads to the recruitment of immune cells, such as T-cells, and orchestration of inflammation. The interaction between the epithelium and T-cells is relatively underexplored in asthma, thus, this study aims to create an in vitro model to investigate the effect of mediators released from viral-infected healthy and asthma bronchial epithelial cells on T-cell activation and differentiation.
Methods: Jurkat cells were stimulated with phorbol-12-myristate-13-acetate (PMA), poly(I:C) and rhinovirus (RV16) to investigate their responsiveness to common immune triggers. In addition, their ability to induce Th1/Th2 cytokine expression was investigated by co-stimulation with CD3/CD28. Finally, Jurkat cells were stimulated with mediators from RV16-infected bronchial epithelial cells from healthy individuals and asthmatic patients to study crosstalk between epithelial cells and T-cells. Gene expression analysis was performed by RT-qPCR, and protein analysis was measured by Western blot and ELISA.
Results: Jurkat cells responded to immune triggers with induction of IL-8, TNFα, IL-2, IFNγ, IL-13 and IL-5. In addition, Jurkat cells could be primed to Th1/Th2 phenotypes using co-stimulation with CD3/CD28. Furthermore, RV16 induced IFNγ, IL-13, and IL-2. Lastly, viral infection-induced bronchial epithelial mediators stimulated cytokine expression in Jurkat cells differentially between healthy and asthma epithelium.
Conclusion: Jurkat cells show plasticity to common immune triggers and respond to signals from the bronchial epithelium. This T-cell response may potentially differ between healthy and asthma epithelium which suggests that further investigation of the crosstalk between epithelium and T-cells could help us better understand disease mechanisms and response to treatment. (Less) - Popular Abstract
- The bronchial epithelium, an immune carpet of the lung that shapes cellular crosstalk
The respiratory system is constantly exposed to various environmental triggers, such as viruses, allergens and pollution. In conditions like asthma, a chronic respiratory disease that affects the airways, these triggers can cause aberrant immune responses, leading to inflammation, coughing and difficulty breathing. The immune system, which protects the body from harmful invaders like viruses, plays an important role in this process. In the lungs, bronchial epithelial cells, which line the airways, are the first line of defense against foreign pathogens. When infected with viruses like the rhinovirus (cause of the common cold), these cells release... (More) - The bronchial epithelium, an immune carpet of the lung that shapes cellular crosstalk
The respiratory system is constantly exposed to various environmental triggers, such as viruses, allergens and pollution. In conditions like asthma, a chronic respiratory disease that affects the airways, these triggers can cause aberrant immune responses, leading to inflammation, coughing and difficulty breathing. The immune system, which protects the body from harmful invaders like viruses, plays an important role in this process. In the lungs, bronchial epithelial cells, which line the airways, are the first line of defense against foreign pathogens. When infected with viruses like the rhinovirus (cause of the common cold), these cells release chemical signals called mediators which communicate with immune cells. However, how these mediators affect T-cells remains unclear, particularly in asthmatic patients. T-cells, key players in the immune system, help the body fight infections but can sometimes overreact, causing excessive inflammation. In this study we focus on investigating this "crosstalk" between bronchial epithelial cells and T-cells to better understand immune responses in both healthy individuals and those with asthma.
The primary goal of this project is to create a model that simulates the interaction between bronchial epithelial cells and T-cells. To do this, we used mediators released by bronchial epithelial cells after they have been infected with rhinovirus. These mediators are then used to stimulate Jurkat cells, a type of T-cell line, to study how these immune cells react to the stimulus. Additionally, we compared how Jurkat cells respond to mediators released from healthy individuals compared to those released from asthmatic patients to see if Jurkat cells respond differently in these conditions.
Overall, our model showed that mediators released from bronchial epithelial cells affect T-cell activation and behavior. Notably, we observed that T-cell responses varied between healthy individuals and asthma patients, indicating possible distinct immunological responses to viral infections.
This experimental model, while needing further refinement, may offer valuable insights into the interplay between innate and adaptive immunity in asthma. By revealing how T-cells behave differently in healthy versus asthmatic individuals, it could help us better understand disease mechanisms. As there is no cure for asthma, this model could be further used to explore immune mechanisms of drug interactions in patients with asthma, potentially leading to improved therapies and treatments.
Master’s Degree Project in Molecular Biology, 60 credits, 2024
Department of biology, Lund University
Advisor: Lena Uller and Mandy Menzel
Biomedical Center, D12, Lund University (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9185210
- author
- Burguet Cavalcanti, Yeilse
- supervisor
-
- Lena Uller LU
- Mandy Menzel LU
- organization
- course
- MOBN03 20232
- year
- 2024
- type
- H2 - Master's Degree (Two Years)
- subject
- language
- English
- id
- 9185210
- date added to LUP
- 2025-02-14 10:27:46
- date last changed
- 2025-02-14 10:27:46
@misc{9185210,
abstract = {{Introduction: Asthma, a heterogenous respiratory disease that affects 300 million people, is characterized by coughing, chest tightness, airway inflammation, and shortening of breath. Asthma exacerbations are commonly triggered by viral infection, such as rhinovirus infection. Respiratory viral infections primarily target the epithelium resulting in the release of alarmins and proinflammatory cytokines. This leads to the recruitment of immune cells, such as T-cells, and orchestration of inflammation. The interaction between the epithelium and T-cells is relatively underexplored in asthma, thus, this study aims to create an in vitro model to investigate the effect of mediators released from viral-infected healthy and asthma bronchial epithelial cells on T-cell activation and differentiation.
Methods: Jurkat cells were stimulated with phorbol-12-myristate-13-acetate (PMA), poly(I:C) and rhinovirus (RV16) to investigate their responsiveness to common immune triggers. In addition, their ability to induce Th1/Th2 cytokine expression was investigated by co-stimulation with CD3/CD28. Finally, Jurkat cells were stimulated with mediators from RV16-infected bronchial epithelial cells from healthy individuals and asthmatic patients to study crosstalk between epithelial cells and T-cells. Gene expression analysis was performed by RT-qPCR, and protein analysis was measured by Western blot and ELISA.
Results: Jurkat cells responded to immune triggers with induction of IL-8, TNFα, IL-2, IFNγ, IL-13 and IL-5. In addition, Jurkat cells could be primed to Th1/Th2 phenotypes using co-stimulation with CD3/CD28. Furthermore, RV16 induced IFNγ, IL-13, and IL-2. Lastly, viral infection-induced bronchial epithelial mediators stimulated cytokine expression in Jurkat cells differentially between healthy and asthma epithelium.
Conclusion: Jurkat cells show plasticity to common immune triggers and respond to signals from the bronchial epithelium. This T-cell response may potentially differ between healthy and asthma epithelium which suggests that further investigation of the crosstalk between epithelium and T-cells could help us better understand disease mechanisms and response to treatment.}},
author = {{Burguet Cavalcanti, Yeilse}},
language = {{eng}},
note = {{Student Paper}},
title = {{Effect of bronchial epithelial mediators on T-cell differentiation and activation}},
year = {{2024}},
}