Abstract / Description of output
Background Gastrointestinal nematode infection is associated with mucosal mast cell (MMC)
hyperplasia. In the mouse, this is accompanied by the release of substantial quantities of the chymase
mouse mast cell proteinase-1 (mMCP-1) into the gut lumen and peripheral bloodstream. Expression
of mMCP-1 is largely restricted to intraepithelial MMC and is thought to play a role in the
regulation of epithelial permeability. MMCs also express mouse mast cell proteinase-2 (mMCP-2),
but less is known about the expression or biological function of this proteinase.
Objectives (1) To purify and characterize mMCP-2. (2) To compare the expression and release of
mMCP-2 and mMCP-1 in vivo using specific antibodies.
Methods Bone marrow-derived mast cells (mBMMCs) were generated from mMCP-1
/
BALB/c
mice. mMCP-2 was purified, characterized and used to generate rat and sheep polyclonal antibodies.
The expression and systemic release of mMCP-1 and -2 were compared in vivo by
immunohistochemistry and ELISA.
Results mMCP-2 was successfully purified from mMCP-1
/
mBMMC and its identity confirmed
by N-terminal amino acid sequencing. mMCP-2 bound [3H]-labelled DFP, indicating the presence of
an active serine proteinase catalytic site, but showed little evidence of chymotryptic activity. MMC
expressed comparable levels of mMCP-1 and -2 in the jejunum but not in the gastric mucosa, where
mMCP-2 was more abundant. Expression of both proteinases increased substantially during primary
Nippostrongylus brasiliensis infection and this was accompanied by a substantial increase in
peripheral blood levels of mMCP-1 (70 mg/mL on day 12). By contrast, mMCP-2 was not detected in
the serum of uninfected mice and only increased to approximately 25 ng/mL on day 12.
Conclusion As in the case of mMCP-1, mMCP-2 expression is restricted to MMC. However,
mMCP-2 lacks chymase activity, is expressed at higher levels in gastric MMC and appears to be
differentially released into the peripheral bloodstream
hyperplasia. In the mouse, this is accompanied by the release of substantial quantities of the chymase
mouse mast cell proteinase-1 (mMCP-1) into the gut lumen and peripheral bloodstream. Expression
of mMCP-1 is largely restricted to intraepithelial MMC and is thought to play a role in the
regulation of epithelial permeability. MMCs also express mouse mast cell proteinase-2 (mMCP-2),
but less is known about the expression or biological function of this proteinase.
Objectives (1) To purify and characterize mMCP-2. (2) To compare the expression and release of
mMCP-2 and mMCP-1 in vivo using specific antibodies.
Methods Bone marrow-derived mast cells (mBMMCs) were generated from mMCP-1
/
BALB/c
mice. mMCP-2 was purified, characterized and used to generate rat and sheep polyclonal antibodies.
The expression and systemic release of mMCP-1 and -2 were compared in vivo by
immunohistochemistry and ELISA.
Results mMCP-2 was successfully purified from mMCP-1
/
mBMMC and its identity confirmed
by N-terminal amino acid sequencing. mMCP-2 bound [3H]-labelled DFP, indicating the presence of
an active serine proteinase catalytic site, but showed little evidence of chymotryptic activity. MMC
expressed comparable levels of mMCP-1 and -2 in the jejunum but not in the gastric mucosa, where
mMCP-2 was more abundant. Expression of both proteinases increased substantially during primary
Nippostrongylus brasiliensis infection and this was accompanied by a substantial increase in
peripheral blood levels of mMCP-1 (70 mg/mL on day 12). By contrast, mMCP-2 was not detected in
the serum of uninfected mice and only increased to approximately 25 ng/mL on day 12.
Conclusion As in the case of mMCP-1, mMCP-2 expression is restricted to MMC. However,
mMCP-2 lacks chymase activity, is expressed at higher levels in gastric MMC and appears to be
differentially released into the peripheral bloodstream
| Original language | English |
|---|---|
| Journal | Clinical & Experimental Allergy |
| DOIs | |
| Publication status | Published - 15 Jul 2003 |