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 ?