Journal List > J Bacteriol Virol > v.41(3) > 1034001

Lee, Jung, Park, Ha, Oh, Oh, and Chai: Proteomic Analysis of the Serum from Chicken Infected by Avian Influenza Virus

Abstract

Avian influenza (AI) is an infectious, low pathogenic virus that is endemic all over the world and poses a potential threat to the poultry industry. Vaccination is a widely used effective method to prevent avian influenza virus. Here we employed a comparative proteomics approach [two-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight (MALDI-TOF)] to characterize proteome in the sera from the specific pathogen free (SPF) chickens, the vaccinated chickens, and the naturally infected chickens. We identified total 58 proteins that were differentially expressed in the sera of three groups. Among them ovotransferrin and vitamin D-binding protein were more expressed in the sera of naturally infected chickens compare with other groups. Our results suggested that the level of these two proteins in the serum may help to discriminate the naturally infected chicken from the vaccinated chicken.

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Figure 1.
A representative result of the analysis of the protein expression in the SPF chicken serum (18 week) and the vaccinated chicken serum (18 week). 100 μg of proteins from the SPF chicken serum (A) and the vaccinated chicken serum (B) were separated by IEF using linear pH gradient strip of pH 4~7 and 12% SDS-PAGE gels. The numbers represent unique numbers automatically assigned to each protein spot by PDQuest2D analysis software.
jbv-41-195f1.tif
Figure 2.
A representative result of the analysis of the protein expression in the SPF chicken serum (7~8 week) and the infected chicken serum (7~8 week). 100 μg of proteins from the SPF chicken serum (A) and the infected chicken serum (B) were separated by IEF using linear pH gradient strip of pH 4~7 and 12% SDS-PAGE gels. The numbers represent unique numbers automatically assigned to each protein spot by PDQuest 2D analysis software.
jbv-41-195f2.tif
Figure 3.
Overexpressed proteins in the infected chicken serum compare with the SPF chicken serum. (A) Ovotransferrin. (B) Vitamin D-binding protein (DBP).
jbv-41-195f3.tif
Table 1.
Differentially expressed proteins list between the SPF chicken serum and the vaccinated chicken serum
Protein Intensity Est'd Z Access number
SPF Vaccine
LOC396098 B6.1 4379.85 0.1 IPI00593934.1
C16orf68 45 kDa protein 1609.15 0.35 IPI00819173.1
LRRIQ1 48 kDa protein 1707.05 2.43 IPI00601234.3
C16orf73 30 kDa protein 2958.73 0.36 IPI00599240.3
HPGD similar to 15-hydroxyprostaglandin dehydrogenase 3674.53 0.92 IPI00580908.3
LOC769732 hypothetical protein, partial 5151.23 0.11 IPI00810964.1
RRP15 hypothetical protein 5368.16 2784.2 0.53 IPI00572505.1
IGLL1 24 kDa protein 13365.83 0.74 IPI00685019.2
PRL prolactin 1404.5 213.13 0.49 IPI00598937.1
FDXACB1 hypothetical protein 21852.3 12805.26 0.33 IPI00591115.2
COMMD2 similar to My004 protein isoform 3 4262.13 0.35 IPI00574181.1
CASP6 caspase-6 1615.5 0.25 IPI00579950.1
LOC396098 26 kDa protein 3589.73 0.55 IPI00819723.1
RCJMB04_1d11 epimorphin 4519.5 0.46 IPI00596666.3
LOC426947 similar to HBV pX associated protein-8; XAP-8, partial 2917.8 1.06 IPI00680167.2
LOC396098 26 kDa protein 8373.77 44518.2 0.18 IPI00819723.1
TSN translin 1566.93 10864.87 0.18 IPI00598812.3
RCJMB04_22d6 abhydrolase domain-containing protein FAM108B1 1269.4 0.86 IPI00594716.3
LOC777249 hypothetical protein, partial 150.2 3509.63 1.43 IPI00811371.1
PPARG peroxisome proliferator-activated receptor gamma 9933.6 1.15 IPI00598191.1
B3GALTL similar to beta3-glycosyltransferase-like; B3GTL 1709.6 5002.4 1.06 IPI00599548.2
PARK7 protein DJ-1 1640.53 12418.6 0.19 IPI00600709.1
CASP3 caspase-3 3632.33 0.34 IPI00584152.1
GSTT1 similar to glutathione S-transferase theta 1 isoform 2 2206.4 0.76 IPI00811441.1
ASNS asparagine synthetase [glutamine-hydrolyzing] 2445.53 24198.17 0.41 IPI00575723.5
PSTK similar to phosphoseryl-tRNA kinase 5217.4 13670.73 0.18 IPI00683380.2
MID1 midline 1 2529.77 7777.03 2.43 IPI00681019.2
DNPEP putative uncharacterized protein 444.2 4913.77 0.41 IPI00587557.1
ND1 NADH-ubiquinone oxidoreductase chain 1 4766.05 0.16 IPI00580952.1

Est'd Z score is a chance value used in ProFound that corresponds to the percentile of the search in a random match population. For instance, a Z score of 1.65 for a search indicates that the search is in the 95th percentile. In the intensity, a dash (–) means that the protein is not expressed in the group.

Table 2.
Differentially expressed proteins list between the SPF chicken serum and the infected chicken serum
Protein Intensity Est'd Z Access number
SPF Infected
31 kDa protein 2640.9 53.7 1.08 IPI00822140.1
LOC416235 ovoinhibitor 2054.9 219.65 0.94 IPI00587313.2
CD14 CD14 antigen 18741.7 7301 0.67 IPI00574630.3
TCF3 Helix-loop-helix protein E47 5909.7 1.85 IPI00822789.1
LOC429793 40 kDa protein 4833.2 258.55 0.56 IPI00820555.1
APOA1 apolipoprotein A-I 4793.3 2513.1 1.76 IPI00580765.1
LOC426508 hypothetical protein 3344.5 1684.2 0.97 IPI00822139.1
LOC771049 similar to integrin alpha-6 precursor (VLA-6) (CD49f antigen), partial 3097.8 803 0.57 IPI00813320.1
MYH15 myosin heavy chain, cardiac muscle (Fragment) 2584.8 0.60 IPI00573338.3
PHB prohibitin 2332.5 0.67 IPI00574627.1
JMJD7-PLA2G4B JMJD7-PLA2G4B protein 649.8 0.53 IPI00886686.1
PSAT1 hypothetical protein 540.5 0.56 IPI00575437.3
IGLL1 24 kDa protein 399.5 1.13 IPI00685019.2
TF ovotransferrin 5401.4 1.07 IPI00683271.1
GC vitamin-D binding protein 2386.8 2.31 IPI00573327.1

Est'd Z score is a chance value used in ProFound that corresponds to the percentile of the search in a random match population. For instance, a Z score of 1.65 for a search indicates that the search is in the 95th percentile. In the intensity, a dash (–) means that the protein is not expressed in the group.

Table 3.
Differentially expressed proteins list between the vaccinated chicken serum and the infected chicken serum
Protein Intensity Est'd Z Access number
Vaccinated Infected
TPM3 isoform 2 of tropomyosin beta chain 18905.45 2.43 IPI00602216.1
APOA4 apolipoprotein AIV 152.6 1.67 IPI00579242.1
UQCRC1 similar to ubiquinol —cytochrome c reductase 1793.6 1.39 IPI00571288.3
CD72 C-type lectin short form 2236.6 0.84 IPI00681573.1
KIAA1467 uncharacterized protein KIAA1467 homolog 4193.5 0.52 IPI00602773.2
LOC771978 similar to pol, partial 3104.55 1.2 IPI00812453.1
LOC429793 40 kDa protein 258.55 0.56 IPI00820555.1
PIT 54 PIT 54 3479.75 0.5 IPI00684336.1
NANS 40 kDa protein 1584.15 0.52 IPI00818823.1
TMPO 28 kDa protein 2173.55 2.43 IPI00822966.1
ATP6V1C1 similar to vacuolar ATP synthase subunit C 2933.45 0.98 IPI00579791.3
C16orf79 hypothetical protein 1171.5 0.9 IPI00811960.1
LOC423849 46 kDa protein 4596.65 0.83 IPI00821041.1
ACTA1 actin, alpha skeletal muscle 1297.4 0.67 IPI00582169.1
GC vitamin-D binding protein 2386.8 2.31 IPI00573327.1
TF ovotransferrin 5401.4 1.07 IPI00683271.1

Est'd Z score is a chance value used in ProFound that corresponds to the percentile of the search in a random match population. For instance, a Z score of 1.65 for a search indicates that the search is in the 95th percentile. In the intensity, a dash (–) means that the protein is not expressed in the group.

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