Development and characterization of ex vivo inflammatory Mannan-Induced Psoriasis model using Ncf1 mutated mice in a 3D nanofiber cell culture system
(2021) MOBN03 20202Degree Projects in Molecular Biology
- Abstract
- Autoimmune diseases are complex and characterized by the loss of self-tolerance to self-antigens, leading to an immune attack on target tissues of the body. Psoriasis (Ps) and psoriasis arthritis (PsA) are poorly understood autoimmune diseases with unknown genetic and environmental sources. It has been previously demonstrated that a single intra peritoneal injection of mannan induces an acute inflammation resembling human Ps and PsA-like disease in inbred mouse strains. Consequently, experimental animal models must closely resemble the mechanisms leading to disease in patients and traditional in vitro models have major limitations in predictability when comparing to the in vivo effects. By applying 3D cell culture systems that mimic... (More)
- Autoimmune diseases are complex and characterized by the loss of self-tolerance to self-antigens, leading to an immune attack on target tissues of the body. Psoriasis (Ps) and psoriasis arthritis (PsA) are poorly understood autoimmune diseases with unknown genetic and environmental sources. It has been previously demonstrated that a single intra peritoneal injection of mannan induces an acute inflammation resembling human Ps and PsA-like disease in inbred mouse strains. Consequently, experimental animal models must closely resemble the mechanisms leading to disease in patients and traditional in vitro models have major limitations in predictability when comparing to the in vivo effects. By applying 3D cell culture systems that mimic original biological structures found in cells in vivo, it has allowed limitations to be reduced, provided better predictability and reduced the number of drug candidates. Hence, we developed an ex vivo assay in which skin inflammatory cells from Ps diseased mice ears are isolated through enzymatic and mechanical digestion for cell culturing in 3D systems. Within the developed ex vivo assay, we found that 3D cell cultures were favorable and provided a better effect compared to 2D cell cultures. The 3D cell cultures provided better comparable CD4+ and CD8+ T cell populations, high TNF-α and IL-17A levels and successful evaluation of proliferation in psoriatic lesioned mice ears. Thus, we can suggest that mannan induced psoriasis in B6.Q. Ncf1m1j/m1j mice led to a CD4+ T cell IL-17A driven response that upregulates inflammation. We stress the importance of reproducing the assay when investigating pathogenesis of psoriasis further and using the following protocol and results as a basis for analyzing and evaluating the effects of reference drugs in Ps and PsA in 3D cell cultures. (Less)
- Popular Abstract
- Silvery scaly skin – what makes psoriasis begin?
The long-term skin disease, psoriasis, causes dry, itchy, scaly looking patches with no available cure and the chronic inflammation affects approximately 2-3% of the world’s population, yet the trigger of the disease remains enigmatic. Through the use of animal models, we can gain a deeper understanding of the underlying mechanisms and discover therapeutic treatments.
Our immune system works like a personal bodyguard. It constantly protects us from invaders like viruses and bacteria. Imagine if the immune system’s bodyguard malfunctions and starts attacking our own cells instead of foreign bodies, what would happen? An autoimmune disease would be triggered. Psoriasis and psoriasis... (More) - Silvery scaly skin – what makes psoriasis begin?
The long-term skin disease, psoriasis, causes dry, itchy, scaly looking patches with no available cure and the chronic inflammation affects approximately 2-3% of the world’s population, yet the trigger of the disease remains enigmatic. Through the use of animal models, we can gain a deeper understanding of the underlying mechanisms and discover therapeutic treatments.
Our immune system works like a personal bodyguard. It constantly protects us from invaders like viruses and bacteria. Imagine if the immune system’s bodyguard malfunctions and starts attacking our own cells instead of foreign bodies, what would happen? An autoimmune disease would be triggered. Psoriasis and psoriasis arthritis are poorly understood autoimmune diseases that trigger inflammation in the skin and joints, in which approximately 25% of psoriasis patients develop psoriasis arthritis.
To investigate the cause of the disease, an injection of Mannan, a carbohydrate found in yeast has demonstrated sudden psoriasis-like inflammation in inbred mouse strains. The increased inflammation is observed on diseased mice ears, as seen in Figure 1, in which inflammation occurs by thickening of the inner layer of the ear (black arrow). Animal models provide a better understanding of the complex interactions of immune cells in human psoriasis. By applying 3D cell culture systems that mimic the original biological structures the cell resides in, the number of animals can be reduced thus providing better predictability.
3D cell culture systems – a powerful tool in inflammatory model research
The triggers of psoriasis and psoriasis arthritis are still relatively unknown, however, through the development of a robust method, tissues from inbred mouse strains with psoriasis-like inflammation can be analyzed and evaluated through the activated T cell immune response. In this project, we developed an enzymatic and mechanical digestion protocol to isolate the inflammatory skin cells from the mouse ears for cell culturing in 3D culture systems to analyze the immune response.
Upon evaluation, we found that 3D cell culture systems provided better results of the psoriasis inflammatory immune response in which viable T cell populations were obtained with increased levels of proinflammatory cytokines. Thus, 3D systems can be used for testing of a variety of novel drugs with different doses and parameters, investigating the various mechanisms of the disease and for discovery and development of therapeutic treatments.
Master’s Degree Project in Molecular Biology, 60 credits, 2021
Department of Biology, Lund University
Advisor: Malin Hultqvist Hopkins, Redoxis AB and Maximilian Ottosson, Cellevate AB (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9055421
- author
- Niskala, Alisha
- supervisor
- organization
- course
- MOBN03 20202
- year
- 2021
- type
- H2 - Master's Degree (Two Years)
- subject
- language
- English
- id
- 9055421
- date added to LUP
- 2021-06-16 14:13:30
- date last changed
- 2021-06-16 14:13:30
@misc{9055421,
abstract = {{Autoimmune diseases are complex and characterized by the loss of self-tolerance to self-antigens, leading to an immune attack on target tissues of the body. Psoriasis (Ps) and psoriasis arthritis (PsA) are poorly understood autoimmune diseases with unknown genetic and environmental sources. It has been previously demonstrated that a single intra peritoneal injection of mannan induces an acute inflammation resembling human Ps and PsA-like disease in inbred mouse strains. Consequently, experimental animal models must closely resemble the mechanisms leading to disease in patients and traditional in vitro models have major limitations in predictability when comparing to the in vivo effects. By applying 3D cell culture systems that mimic original biological structures found in cells in vivo, it has allowed limitations to be reduced, provided better predictability and reduced the number of drug candidates. Hence, we developed an ex vivo assay in which skin inflammatory cells from Ps diseased mice ears are isolated through enzymatic and mechanical digestion for cell culturing in 3D systems. Within the developed ex vivo assay, we found that 3D cell cultures were favorable and provided a better effect compared to 2D cell cultures. The 3D cell cultures provided better comparable CD4+ and CD8+ T cell populations, high TNF-α and IL-17A levels and successful evaluation of proliferation in psoriatic lesioned mice ears. Thus, we can suggest that mannan induced psoriasis in B6.Q. Ncf1m1j/m1j mice led to a CD4+ T cell IL-17A driven response that upregulates inflammation. We stress the importance of reproducing the assay when investigating pathogenesis of psoriasis further and using the following protocol and results as a basis for analyzing and evaluating the effects of reference drugs in Ps and PsA in 3D cell cultures.}},
author = {{Niskala, Alisha}},
language = {{eng}},
note = {{Student Paper}},
title = {{Development and characterization of ex vivo inflammatory Mannan-Induced Psoriasis model using Ncf1 mutated mice in a 3D nanofiber cell culture system}},
year = {{2021}},
}