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 ?