PMID:20709948

Citation	Karlstetter, M, Walczak, Y, Weigelt, K, Ebert, S, Van den Brulle, J, Schwer, H, Fuchshofer, R and Langmann, T (2010) The novel activated microglia/macrophage WAP domain protein, AMWAP, acts as a counter-regulator of proinflammatory response. J. Immunol. 185:3379-90
Abstract	Microgliosis is a common phenomenon in neurodegenerative disorders, including retinal dystrophies. To identify candidate genes involved in microglial activation, we used DNA-microarray analysis of retinal microglia from wild-type and retinoschisin-deficient (Rs1h(-/Y)) mice, a prototypic model for inherited retinal degeneration. Thereby, we cloned a novel 76 aa protein encoding a microglia/macrophage-restricted whey acidic protein (WAP) termed activated microglia/macrophage WAP domain protein (AMWAP). The gene consists of three exons and is located on mouse chromosome 11 in proximity to a chemokine gene cluster. mRNA expression of AMWAP was detected in microglia from Rs1h(-/Y) retinas, brain microglia, and other tissue macrophages. AMWAP transcription was rapidly induced in BV-2 microglia upon stimulation with multiple TLR ligands and IFN-gamma. The TLR-dependent expression of AMWAP was dependent on NF-kappaB, whereas its microglia/macrophage-specific transcription was regulated by PU.1. Functional characterization showed that AMWAP overexpression reduced the proinflammatory cytokines IL-6 and IL-1beta and concomitantly increased expression of the alternative activation markers arginase 1 and Cd206. Conversely, small interfering RNA knockdown of AMWAP lead to higher IL-6, IL-1beta, and Ccl2 transcript levels, whereas diminishing arginase 1 and Cd206 expression. Moreover, AMWAP expressing cells had less migratory capacity and showed increased adhesion in a trypsin-protection assay indicating antiserine protease activity. In agreement with findings from other WAP proteins, micromolar concentrations of recombinant AMWAP exhibited significant growth inhibitory activity against Escherichia coli, Pseudomonas aeruginosa, and Bacillus subtilis. Taken together, we propose that AMWAP is a counter-regulator of proinflammatory microglia/macrophage activation and a potential modulator of innate immunity in neurodegeneration.
Links	PubMed Online version:10.4049/jimmunol.0903300
Keywords	Amino Acid Sequence; Animals; Anti-Inflammatory Agents, Non-Steroidal/contraindications; Anti-Inflammatory Agents, Non-Steroidal/metabolism; Anti-Inflammatory Agents, Non-Steroidal/pharmacology; Base Sequence; Cell Adhesion Molecules/deficiency; Cell Adhesion Molecules/genetics; Cell Line; Cells, Cultured; Eye Proteins/genetics; Inflammation Mediators/contraindications; Inflammation Mediators/metabolism; Inflammation Mediators/physiology; Macrophage Activation/immunology; Macrophages/immunology; Macrophages/metabolism; Macrophages/pathology; Mice; Mice, Inbred C57BL; Mice, Knockout; Microglia/immunology; Microglia/metabolism; Microglia/pathology; Milk Proteins/biosynthesis; Milk Proteins/genetics; Milk Proteins/pharmacology; Molecular Sequence Data; Nerve Tissue Proteins/biosynthesis; Nerve Tissue Proteins/genetics; Nerve Tissue Proteins/pharmacology; Neurodegenerative Diseases/immunology; Neurodegenerative Diseases/metabolism; Neurodegenerative Diseases/pathology; Protein Structure, Tertiary; Retina/cytology; Retina/immunology; Retina/metabolism
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
MOUSE:Q5SSJ1		GO:0004867: serine-type endopeptidase inhibitor activity	ECO:0000314:		F		Figure 9E: AMWAP attenuates microglial migration and exerts anti-serine protease activity. Quantitative analysis of anti-protease activity in AMWAP-GFP and GFP expressing BV-2 microglia.	complete CACAO 10071
MOUSE:Q5SSJ1	enables	GO:0004867: serine-type endopeptidase inhibitor activity	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete

References

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