Proteomic profiling of human neutrophils in relation to immunoglobulin G Fc receptor IIIb polymorphism.
OBJECTIVE
Neutrophils are essential in host defense against periodontopathic bacteria. Immunoglobulin G Fc receptor IIIb (FcγRIIIb) is a neutrophil-specific receptor for immunoglobulin G and bears the functional NA1-NA2 polymorphism. Accumulating evidence suggests a significant association between FcγRIIIb gene polymorphism and periodontitis. In this study, we employed a proteomic approach to evaluate the relevance of FcγRIIIb polymorphism to protein expression profiles of neutrophils.


METHODS
Neutrophils were collected from ten healthy subjects whose FcγRIIIb genotypes were determined by allele-specific PCRs. Expressions of proteins induced by interaction via FcγRIIIb were examined between the FcγRIIIb genotypes with two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis. Proteins that were significantly different in expression levels between the FcγRIIIb genotypes were determined with computer image analysis, and identified with mass spectrometry and protein databases.


RESULTS
A total of 757 protein spots were observed in the two-dimensional electrophoretograms of neutrophils from five FcγRIIIb-NA1/NA1 and five FcγRIIIb-NA2/NA2 donors. A statistical analysis revealed that the expression levels of five proteins were significantly different between the FcγRIIIb genotypes (p < 0.05). The FcγRIIIb-NA1/NA1 neutrophils exhibited two spots that were significantly underexpressed (protein-arginine deiminase type-4 and annexin VI) and three spots that were significantly overexpressed (Cdc42hs-Gdp complex, myosin light chain 12A and coactosin-like 1) when compared with FcγRIIIb-NA2/NA2 neutrophils. The same expression profiles of protein-arginine deiminase type-4 were obtained by ELISA.


CONCLUSIONS
Differential protein expression profiles were observed in neutrophils between FcγRIIIb genotypes.
Yokoyama
T
T
Division of Periodontology, Department of Oral Biological Science, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
Kobayashi
T
T
Yamamoto
K
K
Yamagata
A
A
Oofusa
K
K
Yoshie
H
H
D000328:Adult; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D058851:GPI-Linked Proteins / Q000235:genetics; D020869:Gene Expression Profiling; D005787:Gene Frequency; D006801:Humans; D006867:Hydrolases / Q000235:genetics; D007091:Image Processing, Computer-Assisted; D007519:Isoantigens / Q000235:genetics*; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D009504:Neutrophils / Q000737:chemistry* / Q000378:metabolism; D011110:Polymorphism, Genetic; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases; D020543:Proteome; D017452:Receptors, IgG / Q000235:genetics*; D055815:Young Adult
C428200:FCGR3B protein, human; D058851:GPI-Linked Proteins; D007519:Isoantigens; D020543:Proteome; D017452:Receptors, IgG; C067704:neutrophil-specific antigen NA1, human; C072753:neutrophil-specific antigen NA2; D006867:Hydrolases; C000629235:PADI4 protein, human; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases