Transcript CHRONIC OBSTRUCTIVE PULMONARY DISEASE

CHRONIC OBSTRUCTIVE
PULMONARY DISEASE
ZULEYHA OZEN
OVERVIEW
● Introduction
● Information about Chronic Obstructive Pulmonary
Disease (COPD)
● Inflammatory Responses
● Paper 1
● Paper 2
● What is still unknown?
● Future Studies/Specific Aim
Disease Overview

Pathophysiology of COPD (Chronic
Bronchitis and Emphysema)
-Chronic Bronchitis: Loss of muco-ciliary
clearance

Loss of cilia function
-Emphysema: Destruction of elastin fibers

Proteases, matrix metalloproteinase 9
(MMP9) cause elastin degradation
Shortness of Breath
Cough/Sputum-(Progressive dyspnea)
http://www.wgabinecie.pl/artykul/65-pochp-przewlekla-obturacyjna-choroba-pluc/
Current existing medicines

Bronchodilators

Combination of Bronchodilators and cortisteroids

Vaccines

Pulmonary Rehabilitation

Oxygen Therapy

Change in Lifestyle
http://www.ncbi.nlm.nih.gov/books/NBK26827/figure/A4531/?report=objectonly
https://www.caymanchem.com/app/template/Article.vm/article/2177
Inflammatory and immune
cells involved in chronic
obstructive pulmonary
disease (COPD)
Macrophages and epithelial
cells:
-chemotactic factors
-attract inflammatory cells
http://www.nature.com/nri/journal/v8/n3/fig_tab/nri2254_F2.html
TH17 cells and airway
inflammation
http://www.nature.com/nri/journal/v8/n3/fig_tab/nri2254_F5.html
Smoke Exposure (n=20/group)
Whole body exposed to cigarette smoke (CS)
20 mice exposed
to CS of 5
cigarettes
30 min
smoke
free
interval
20 mice exposed
to CS of 5
cigarettes
30 min
smokefree
interval
20 mice exposed
to CS of 5
cigarettes
30 min
smokefree
interval
20 mice exposed
to CS of 5
cigarettes
The CS exposure animals were divided into two subgroups:
Control group:
exposed to air
Sub-acute exposure:
Initiated CS at 26-28
weeks old
Mice of each group 30-32 weeks old when
the CS & Air exposure ended
Chronic exposure:
initiated CS at 6-8
weeks old
Figure 1. Assessing effect of CS
exposure on lung inflammation
Total number of alveolar inflammatory cells
increased in chronic CS exposure
•
Sub-acute exposure significantly lower than
chronic CS
Figure 2A. Prevalence of Th17 cells in lung tissue
Prevalence of Th17 cells:
-ratio of CD4+ IL-17A+ cells to the
total amount of CD4+ T lymphocytes
-Th17 prevalence markedly higher in
mice with chronic CS and Sub-acute
CS
Figure 2B. Peripheral blood mononuclear cells
-similarly the prevalence of Th17 cells increased in CS
exposure
Figure 2C. Collective analysis
of flow cytometry
Figure 3A. Prevalence of T regulatory cells (ratio of CD4+CD25+Foxp3+ cells to the
total amount of CD4+ T lymphocytes) in lung tissue
Prevalence of T regulatory cells:
-markedly higher in sub-acute
exposure
-drops in chronic CS exposure
Figure 3B. Peripheral blood mononuclear cells
-similarly the prevalence of T regulatory cells decreased in chronic CS exposure
Figure 3C. Collective analysis
of flow cytometry
Figure 4. Assessing the ratio of Th17 and Treg in lung tissue
and peripheral blood
-In lung tissues, the ratio of Th/Treg
is decreased with sub-acute CS
exposure
-Increases in chronic CS exposure
Ratio of Th17/Treg in peripheral
blood:
-significantly increased in chronic
CS exposure
Figure 5. The expression of Foxp3 and
ROR gamma t mRNA
-Looking at specific transcription factors of both Tsubsets by real time-PCR
-Th17 specific transcription factor ROR gamma t
mRNA expression: significantly increased in CS
exposed
-T regulatory specific transcription factor Foxp3
mRNA expression: significantly decreased in CS
exposed
Table 1: levels of IL-17A, IL-6, IL23 and TGF-beta in serum
significantly higher in chronic CS
exposure
-IL-10 sig. lower in chronic CS
Conclusions from paper 1

There is an obvious imbalance between Th/Treg cells in CS exposed mice

Prevalence of Th17 and Th17 specific transcription factor, ROR gamma t mRNA:
increased
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Treg cell prevalence and Treg specific transcription factor, Foxp3 mRNA: decreased


Thus, leading to an imbalance in the ratio of Th/Treg cell profiles
The existing cytokine profile can be further evaluated for specific therapeutic
approach
Figure 1. Assessing histology of lung tissues
Fig.1 Histology of Lung Tissues
●
Mean alveoli were expanded and broken
●
COPD lung- more inflammatory cells
Figure 2. Expression of transcription factors ROR gamma t
and Foxp3
•
•
•
Fig.2a. Foxp3 relative mRNA expression level significantly lower in COPD patients
Fig.2b. RORyt relative mRNA expression level significantly higher in COPD patients
Fig.2c. Ratio of Treg/Thelper cells in the level of mRNA lower in COPD patients
Figure 3. Assessing expression of Foxp3 and ROR gamma t protein levels
-Increased protein
expression of ROR gamma t
in COPD patients
-Viceversa, decreased
protein expression of Foxp3
compared with smokers and
nonsmokers
Figure 4. Immunohistochemistry staining
of different proteins
All p-values were less than 0.001
-IL-17+, CCR6+ and IL-23R cells increased
-Foxp3 cells in alveolar walls decreased
Figure 5. The number of Foxp3+,
IL-17+, CCR6+ and IL-23R+ cells in
alveolar walls
-Foxp3+ and Foxp3+ / IL-17+ cells
decreased in cell number in chronic CS
exposure
-IL-17+, CCR6+ and IL-23R+ cells
increased in cell number in chronic CS
exposure
-Foxp3+ & Foxp3+/IL-17+ cells decreased
in alveolar walls of COPD
A: ROR gamma t & Foxp3 mRNA expression: negatively correlated
B: Ratio of Foxp3/ROR gamma t mRNA expression negatively correlated with mean alveoli area
C: Positive correlation with the ratio of Foxp3/ROR gamma t and FEV1%pred
D: ROR gamma t & Foxp3 protein: negatively correlated
E: Ratio of Foxp3/ROR gamma t in level of protein and mean alveoli area
F: Positive correlation with the ratio of Foxp3/ROR gamma t in level of protein and FEV1%pred
G: Numbers of Foxp3+/IL-17+ cells: negatively correlated
H: Ratio of Foxp3/IL-17+ cells: negatively correlated to the mean alveoli area
I: Positive correlation with the ratio of Foxp3+/IL-17+ cells and FEV1%pred
Conclusions from paper 2

Decreased ratio of Foxp3/ROR gamma t in patients with COPD and normal smokers


persistent with the aggravation of the disease
Decreased ratio of Foxp3/ROR gamma t important in pathogenesis of COPD
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Immune dysregulation, and participation in lung inflammation: leading to destruction in
the lung

Pro-inflammatory cytokines and chemokine receptors are also evident in development of
the disease

Their association with the transcription factors ROR gamma t and Foxp3 can be further
researched for potential therapeutics
What is still unknown ?

The regulatory cytokine involvement: TGF-beta levels in the progression of the
disease is still unknown

Study by Zhou et al. TGF-beta induced Foxp3 leads to inhibition of Th17 cell
differentiation

In a dose dependent matter, TGF-beta might be a factor seen in the imbalance
between Th/T regulatory cells
Specific Aim

In the research proposal, my area of focus will be to further analyze the negatively correlated
relationship between the T-regulatory and T helper cells with regards to changes in the
expression of the specific transcription factors and cytokine TGF-beta
 Implement TGF-beta induced Foxp3 and use of cytokine IL-6 antagonist for therapeutic
approach
THANK YOU! QUESTIONS?