1 Mouse Models of Inflammatory Lung Diseases Zhou Zhu, MD, Ph.D. Division of Allergy and Clinical Immunology Johns Hopkins University School of Medicine Outlines • Basic lung structure, function, and inflammatory lung diseases • Methods, techniques, and approaches to studying lung diseases • Mouse models of inflammatory lung diseases From Wikipedia Lung: Basic Structure Anatomy
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Mouse Models of Pulmonary Inflammation · OVA Induced Airway Inflammation PBS OVA BAL Cells Macrophages Eosinophils. 4 OVA Induced Airway Inflammation P S /T T 0 10 20 30 40 BAL Total
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Mouse Models of Inflammatory Lung Diseases
Zhou Zhu, MD, Ph.D.
Division of Allergy and Clinical Immunology Johns Hopkins University School of Medicine
Outlines
• Basic lung structure, function, and inflammatory lung diseases
• Methods, techniques, and approaches to studying lung diseases
• Mouse models of inflammatory lung diseases
From Wikipedia
Lung: Basic Structure
Anatomy
2
Lung: Basic Structure
H&E (100x)Lung Histology
AirwaysBloodVesselAlveoli
Lung Function and Diseases
• Gas exchange: O2 from atmosphere to bloodstream and CO2
Protocol for ovalbumin (OVA) induced allergic asthma
OVA Induced Airway Inflammation
PBS OVA
BAL Cells
Macrophages Eosinophils
4
OVA Induced Airway Inflammation
PBS/WT
OVA/WT
0
10
20
30
40
BA
L To
tal c
ell n
umbe
r (X
104 )
Mac Neu EosLym
0
20
40
60
80
100
120 PBS/WT
OVA/WTce
ll d
iffer
entia
tio
n(%
)
Total Cell Counts Differential Cell Counts
OVA Induced Lung Inflammation
PBS OVA
H&E (100x)
OVA Induced Lung Inflammation
PBS OVA
H&E (400x)
5
OVA Induced Lung Inflammation
PBS OVA
Alcian Blue (100x)
OVA Induced Lung Inflammation
PBS OVA
Alcian Blue (400x)
OVA Induced Lung Inflammation
PBS OVA
Trichrome (100x)
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OVA Induced Lung Inflammation
PBS OVA
Trichrome (400x)
Elastase Induced Emphysema
Protocol for elastase induced emphysema
Day 0
Porcine Pancreatic Elastase
1u/mouse, I.T. once
28
Sacrifice &
Collect BAL, lung tissue
1 2 3 7 14 21
Elastase Induced Airway Inflammation
PBS
BAL Cells
Day 2 Day 21
7
Elastase Induced Emphysema
Lung Histology (H&E 20x)
Day 3 Day 21
SHP-1
Src Homology 2 domain-containing protein tyrosinePhosphatase-1
Encoded by thehcp genePrimarily expressed in hematopoietic-derived cellsNegative regulator of a variety of growth factor/cytokinesignaling pathwaysActivated SHP-1→ binds to its target proteins→ dephos-phorylates the phospho-tyrosine residues→ terminates thesignaling
SHP-1 Regulation of Signaling
Inactive
SH2
SH2 N
C
SH2
SH2
N
C
ITIM
ITIM
Subs
Active
P
P Subs
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SHP-1 Deficiency in Mice
� Motheaten mice (me/me) – a natural mutant strain
� Viable motheaten mice (mev/mev) – another spontaneous mutant strain
⇒ The mutations cause aberrant splicing of the SHP-1 transcript, resulting in diminished production of functional SHP-1 protein
� Phenotypeinflammatory lesions of the skin, increased neutrophils in the peripheral blood, glomerularnephritis, abnormal lymphoid tissues, pneumonitis with macrophage infiltration in the alveoli, increased production of immunoglobulins, depressed immune responses, profound NK-cell deficiency, and a shortened life span (average 22 days and 61 days, respectively)
IL-1
3Rα1
α1
α1α1
IL-4
Rαα αα
IL-4/IL-13
Tyk-2
IL-4/IL-13Responsive GenesS
TAT-
6
IRS-1/2Cell Growth& Survival
Jak-1
γγ γγc
IL-4
Rαα αα
IL-4
PI3K
IL-4/IL-13 Signaling Systems
Type I Type II
SHP-1SHIP-1SOCS
Time (min): 0 3 5 10 30 5 10 30
TNF-αααα IL-13
p- SHP-1
SHP-1
A B C
IHC: Control IgG αααα-SHP-1 αααα-SHP-1
MLE12
Mouse Lung
SHP-1 Expression and Activation in Lung Epithelial Cells
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Hypothesis
SHP-1 is an important regulator of • Immunological homeostasis of the lung• Allergic inflammatory responses