Journal Club

Background
Thermal injury model
Three day progression to ulceration
Skin remodeling and scar forming
IgM or complement dependent process
Mast cell
Degranulation is an early event (less than 2 hours)
Protection with absence of different proteases
mMCP5 and mMCP4







Methods
Mice (8 to 14 wks)
C57BL/6
Mutants
mMCP4 and mMCP5 knockouts
Thermal challenge model
Prior anesthesia
Application of high-impact nonheat-conducting plastic
container (square shaped 1 cm
2
interscapular)
Skin was scalded in water bath at 54C for 30s
Post-burn variations



Neunlist, M. et al. (2012) The digestive neuronalglialepithelial unit: a new actor in gut health and disease
Nat. Rev. Gastroenterol. Hepatol. doi:10.1038/nrgastro.2012.221
Figure 1 Junctional complexes regulating epithelial cells interactions
Figure 1

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Figure 8

Figure 8

Conclusions
Thermal challenge
MC zonal degranulation at 30 s with expansion
at 120 s
Disruption of the TJs suggests an initiating role
for MC in the early loss of claudin 4 and
appearance of epidermal edema
Protection of the TJs in the protease-deficient
strains
Leder Chloracetate esterase:
The slides are incubated in a solution
naphthol AS-D chloroacetate.
Then the esterase contained in the neutrophils and mast
cells binds with the chloroacetate. This releases the
naphthol
group, which binds to the diazonium dye pararosanalin
(basic fuchsin), another component of the incubating
solution.
Pararosanilin or basic fuchsin gives a deep pink-red
color to the granules, while hematoxylin counterstains the
nuclei blue.

Background
Previous MC-deficient Kit
W-sh/W-sh
and
antibody mediated depletion of basophils
experiments
Role in peanut induced anaphylaxis (PIA)
Interpretation of data
W-sh/W-sh other phenotypic abnormalities
Antibodies react with MC




Methods
Mice
WT vs mutants
C57BL/6-Kit
W-sh/W-sh
Mcpt5-Cre;iDTR
Mcpt8
DTR
Cpa3-Cre;Mcl-1
fl/fl
Peanut induced anaphylaxis
Orally sensitized for 4 weeks
10 mg peanut extract
20 g/100 l cholera toxin
Challenged IP 2 weeks later
5mg/200 l peanut extract
Rectal temperature
Immediately
Different time points


Kit
W-sh/W-sh


Mcpt5-Cre;iDTR
Basophils
Mcpt8
DTR

Cpa3-Cre;Mcl-1
fl/fl

Conclusions
Kit
W-sh/W-sh
Decrease in immediate hypothermia
Late decrease between 3-6 hours
Disappears after using anti-Ly6G antibodies
Neutrophil dependent
Mcpt5-Cre;iDTR
Significant reduction of hypothermia
Residual due to MMC
Mcpt8
DTR
Significant reduction of hypothermia
Cpa3-Cre;Mcl-1
fl/fl
Reduced but significant hypothermia
No increase in histamine levels
PAF levels did not decrease that much
MC or basophils
Monocytes or neutrophils