Transcript Group_3_Final_Presentation - Mast Cell

Group 3
Mast cell deficient mice models
COST action BM1007 Training school
Jerusalem
January 2014
The Kit-deficient mouse models for mast cell research
WBB6F1-KitW/KitW-v
• no mast cells
• no melanocytes
MC
reaction
Kit+/+
yes
yes
KitW/KitW-v
no
no
yes
?
MZ
• lack in γδ T cells
• anemic
• neutropenia
MZ
MZ
• sterile
MZ
yes, but deficient
The Kit-deficient mouse models for mast cell research
C57B6-Kitsh/Kitsh (sash)
• no mast cells
MC
reaction
Kit+/+
yes
yes
Kitsh/Kitsh
no
no
yes
?
MZ
• no melanocytes
• neutrophilia
• splenomegaly
MZ
MZ
MZ
yes, but deficient
Available Kit-independent MC-deficient mouse models
Types
Mcpt5-Cre; iDTR
Mcpt5-Cre; RDTA
Mas-TRECK
Cre-Master
Cpa3Cre/+
Hello Kitty
Cpa3-Cre; Mcl1fl/fl
Available
background
Mechanism
MC
affected
Type of MC
deficiency
Treatment
Gene
C57/BL6
DT mediated
depletion of
MCPT5 expressing
cells.
CTMC
Inducible
Injection of
DT
MCPT5
C57/BL6
DT mediated
depletion of
MCPT5 expressing
cells.
CTMC
Constitutive
-
MCPT5
C57/BL6
DT mediated
depletion of cells
that employ IE
enhancer element
to express IL-4.
MTMC/
CTMC
Inducible
Injection of
DT
C57/BL6
Depletion
mediated by toxic
effects of Cre
under the control
of Cpa3
MTMC/
CTMC
Constitutive
-
Cpa3
-
C57/BL6
Depletion by Mcl1 mediated
induced by Cre
expression under
the control of
Cpa3 promoter
MTMC/
CTMC
Constitutive
-
Cpa3
macrocytic anemia
Systemic abnormalities
Cellular abnormalities
General disadventages Level of defficiency
Side effects after injection
Destruction of the local
No report about Basophil
of DT (Local and systemic
tissue due to injection around 2,5% ear skin
numbers in PBMCs
DT injection)
of DT
?
?
Basophils are also
IE enhancer Side effects after injection
depleted. Is that really
element of IL- of DT (250 ng I.P. 5 before
specific of that cell types
4 gene
experiment)
in chronic conditions?
?
Restoring
Phenotype
Time: 3 weeks
around 3,5 ear skin
and 11% in the back Reconstitution ???
and abdominal skin
?
Total depletion
Time: 18 days
Basophils reduction
around 40%
?
Total depletion
Reconstitution
Basophils reduction
around 40%, neutrophilia
?
92 to 100%
Reconstitution
Selective ablation of mast cells or basophils
reduces peanut-induced anaphylaxis in mice
REBER, L. L., ET AL.
CLINICAL AND TRANSLATIONAL ALLERGY (2013)
Mast cell deficient mice models
KitW-sh/W-sh
neutrophil depleted- KitW-sh/W-sh
Mcpt5-Cre+;iDTR+
Basophil deficient mice models
Ba10336 antibody
Mcpt8DTR/+
Mast cell and basophil deficient mice models
Cpa3-Cre;Mcl-1fl/fl
Cre-Mediated Cell Ablation Contests Mast
Cell Contribution in Models of Antibody and
T Cell-Mediated Autoimmunity
FEYERABEND, THORSTEN B., ET AL.
IMMUNITY (2011)
Nippostrongylus brasiliensis
Antibody-Induced Arthritis
Encephalomyelitis
Mast Cells Are Key Promoters of Contact
Allergy that Mediate the Adjuvant Effects of
Haptens
DUDECK, ANNE, ET AL.
IMMUNITY 34.6 (2011)
KitW/Wv
KitW-sh/W-sh
Mcpt5-Cre+;iDTR+
Mcpt5-Cre+R-DTA+
KitW-sh/W-sh neutrophil depleted
Summary
Kit dependent mutants
Kit independent mutants
 KitW/W-v
 Mcpt5-Cre;R-DTA/iDTR
 KitW-sh/W-sh
 “Cre-Master”Cpa3Cre/+
Conclusions
No model is perfect !
Kit dependent mutants
Kit independent mutants
 Non specific immune effects
 Possible effects on other cells
 Effects of apoptosis
 Breeding / economical
 Compensation effect
 Compensation effect
Variable results depending on different animal strains/models,
protocols, disease model etc.
“ Based on the results obtained so far with both the older and newer
models for MC research, we think that the most robust conclusions
about what MCs can do (or don’t do) in various biological responses
in vivo, and regarding the importance of such MC contributions, are
likely to be derived from investigations that employ multiple
informative model systems. This approach increases the cost of such
work, but permits one to exploit the attractive features of the various
models while keeping in mind the known and potential limitations in
each of them. “
Reber, Marichal and Galli, 2012
But still…
Results obtained so far – Valuable information
Availability / cost effective
Take home message:
always ask the right question!
Suggestions for improvements:
Thanks Frank!
Questions ?