Group 3

Mast cell deficient mice models

COST action BM1007 Training

school Jerusalem

January 2014

MC reaction

yesyesKit+/+MZ

nonoKitW/KitW-v

MZ

?yesMZ

• no mast cells• no melanocytes• lack in γδ T cells• anemic• neutropenia• sterile

WBB6F1-KitW/KitW-v

MZ

yes, but deficient

The Kit-deficient mouse models for mast cell research

The Kit-deficient mouse models for mast cell research

MC reaction

yesyesKit+/+

MZ

nonoKitsh/Kitsh

MZ

?yesMZ

•no mast cells•no melanocytes• neutrophilia• splenomegaly

C57B6-Kitsh/Kitsh (sash)

MZ

yes, but deficient

Types Available background Mechanism MC

affectedType of MC deficiency Treatment Gene Systemic

abnormalitiesCellular

abnormalitiesGeneral

disadventagesLevel of

defficiencyRestoring Phenotype

Mcpt5-Cre; iDTR C57/BL6

DT mediated depletion of

MCPT5 expressing cells.

CTMC Inducible Injection of DT MCPT5

Side effects after injection of DT (Local

and systemic DT injection)

No report about Basophil numbers in

PBMCs

Destruction of the local tissue due to

injection of DTaround 2,5% ear

skin Time: 3 weeks

Mcpt5-Cre; R-DTA C57/BL6

DT mediated depletion of

MCPT5 expressing cells.

CTMC Constitutive - MCPT5 ? ? ? around 3,5 ear skin and 11% in the back and abdominal skin

Reconstitution???

Mas-TRECK C57/BL6

DT mediated depletion of cells

that employ IE enhancer element

to express IL-4.

MTMC/ CTMC Inducible Injection of

DTIE enhancer

element of IL-4 gene

Side effects after injection of DT (250 ng

I.P. 5 before experiment)

Basophils are also depleted. Is that really

specific of that cell types in chronic

conditions?? Total depletion Time: 18 days

Cre-Master Cpa3Cre+/ C57/BL6

Depletion mediated by toxic

effects of Cre under the control

of Cpa3

MTMC/ CTMC Constitutive - Cpa3 - Basophils reduction

around 40% ? Total depletion Reconstitution

Hello KittyCpa3-Cre; Mcl-1fl/fl

C57/BL6

Depletion by Mcl-1 mediated

induced by Cre expression under

the control of Cpa3 promoter

MTMC/ CTMC Constitutive - Cpa3 macrocytic anemia

Basophils reduction around 40%, neutrophilia

? 92 to 100% Reconstitution

Available Kit-independent MC-deficient mouse models

Selective ablation of mast cells or basophils reduces peanut-induced anaphylaxis in mice

REBER, L. L., ET AL .CLINICAL AND TRANSLATIONAL ALLERGY (2013)

KitW-sh/W-sh neutrophil depleted- KitW-

sh/W-sh

Mcpt5-Cre+;iDTR+

Mast cell deficient mice models

Basophil deficient mice models

Mcpt8DTR/+Ba10336 antibody

Cpa3-Cre;Mcl-1fl/fl

Mast cell and basophil deficient mice models

Cre-Mediated Cell Ablation Contests Mast Cell Contribution in Models of Antibody andT 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)

Mcpt5-Cre+;iDTR+ Mcpt5-Cre+R-DTA+

KitW/Wv KitW-sh/W-sh KitW-sh/W-sh neutrophil depleted

Summary

KitW/W-v

KitW-sh/W-sh

Kit dependent mutants Kit independent mutants

Mcpt5-Cre;R-DTA/iDTR

“Cre-Master”Cpa3Cre/+

Conclusions No model is perfect !

Kit dependent mutants Kit independent mutants

Non specific immune effects

Compensation effect

Variable results depending on different animal strains/models, protocols, disease model etc.

Possible effects on other cells Effects of apoptosis Breeding / economical Compensation effect

“ 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!

Thanks !

Questions ?