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Evaluation of long-term antibody response and cross-serotype reaction in ducks immunised with recombinant Riemerella anatipestifer outer membrane protein A and CpG ODN

May Phonvisay

May Phonvisay

Department of Tropical Agriculture and International CooperationNeipu, Taiwan
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Phonvisay, May
, 
Jai-Wei Lee

Jai-Wei Lee

Department of Tropical Agriculture and International CooperationNeipu, Taiwan
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Lee, Jai-Wei
, 
Jhong-Jie Liou

Jhong-Jie Liou

Graduate Institute of Animal Vaccine Technology, College of Veterinary Medicine, National Pingtung University of Science and TechnologyNeipu, Taiwan
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Liou, Jhong-Jie
, 
Hsian-Yu Wang

Hsian-Yu Wang

Graduate Institute of Animal Vaccine Technology, College of Veterinary Medicine, National Pingtung University of Science and TechnologyNeipu, Taiwan
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Wang, Hsian-Yu
 und 
Chun-Yen Chu

Chun-Yen Chu

Graduate Institute of Animal Vaccine Technology, College of Veterinary Medicine, National Pingtung University of Science and TechnologyNeipu, Taiwan
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Chu, Chun-Yen
  
16. Nov. 2019
Evaluation of long-term antibody response and cross-serotype reaction in ducks immunised with recombinant Riemerella anatipestifer outer membrane protein A and CpG ODN's Cover Image
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Online veröffentlicht: 16. Nov. 2019
Seitenbereich: 543 - 548
Eingereicht: 10. Apr. 2019
Akzeptiert: 24. Okt. 2019
DOI: https://doi.org/10.2478/jvetres-2019-0066
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© 2019 M. Phonvisay et al. published by Sciendo
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Fig. 1

Expression and immunogenicity analysis of recombinant OmpA. A – rOmpA separated on 10% SDS-PAGE for protein quantitation. Lanes 1–5 – BSA standards, lane 6 – lysate of E. coli expressing rOmpA induced for 4 h, lane 7 – lysate of E. coli expressing inactivated rOmpA, and lane M – gene marker. B – The 65 kDa of expressed rOmpA separated on 10% SDS-PAGE. Lane 1 – pET-32a incubated for 0 h, lane 2 – pET-32a incubated for 4 h, lane 3 – lysate of E. coli expressing rOmpA induced for 0 h, lane 4 – lysate of E. coli expressing rOmpA induced for 4 h, and lane M – gene marker. C – Western blot analysis with anti-RA duck sera. Lane 1 – pET-32a incubated for 0 h, lane 2 – pET-32a incubated for 4 h, lane 3 – lysate of E. coli expressing rOmpA induced for 0 h, lane 4 – lysate of E. coli expressing rOmpA induced for 4 h and M – gene marker
Expression and immunogenicity analysis of recombinant OmpA. A – rOmpA separated on 10% SDS-PAGE for protein quantitation. Lanes 1–5 – BSA standards, lane 6 – lysate of E. coli expressing rOmpA induced for 4 h, lane 7 – lysate of E. coli expressing inactivated rOmpA, and lane M – gene marker. B – The 65 kDa of expressed rOmpA separated on 10% SDS-PAGE. Lane 1 – pET-32a incubated for 0 h, lane 2 – pET-32a incubated for 4 h, lane 3 – lysate of E. coli expressing rOmpA induced for 0 h, lane 4 – lysate of E. coli expressing rOmpA induced for 4 h, and lane M – gene marker. C – Western blot analysis with anti-RA duck sera. Lane 1 – pET-32a incubated for 0 h, lane 2 – pET-32a incubated for 4 h, lane 3 – lysate of E. coli expressing rOmpA induced for 0 h, lane 4 – lysate of E. coli expressing rOmpA induced for 4 h and M – gene marker

Fig. 2

RA-specific antibody level in immunised ducks. Different superscript letters indicate statistically significant difference (p < 0.05) between groups for each sampling time point
RA-specific antibody level in immunised ducks. Different superscript letters indicate statistically significant difference (p < 0.05) between groups for each sampling time point

Fig. 3

Cytokine mRNA levels in immunised ducks. The mRNA expression levels of IFN-α (A), IFN-γ (B), IL-6 (C), and IL-12 (D) are shown as fold change relative to the saline control. Data are presented as the mean ± standard deviation of ducks in the treatment (n = 5). Different superscript letters indicate statistically significant difference (p < 0.05) between groups for the same sampling time point
Cytokine mRNA levels in immunised ducks. The mRNA expression levels of IFN-α (A), IFN-γ (B), IL-6 (C), and IL-12 (D) are shown as fold change relative to the saline control. Data are presented as the mean ± standard deviation of ducks in the treatment (n = 5). Different superscript letters indicate statistically significant difference (p < 0.05) between groups for the same sampling time point

Fig. 4

Cross-serotype detection by antiserum from ducks immunised with rOmpA + CpG. A – Detection with antiserum of unimmunised ducks. B – Detection with antiserum of ducks immunised with rOmpA + CpG. RA1 – RA serotype 1, RA2 – RA serotype 2, RA6 – RA serotype 6, RA56a – RA of unknown serotype, rOmpA – rOmpA induced for 4 h, and M – protein ladder with labelled molecular weights. The positions of the 65-kDa rOmpA and the 42-kDa OmpA proteins are indicated by the arrows
Cross-serotype detection by antiserum from ducks immunised with rOmpA + CpG. A – Detection with antiserum of unimmunised ducks. B – Detection with antiserum of ducks immunised with rOmpA + CpG. RA1 – RA serotype 1, RA2 – RA serotype 2, RA6 – RA serotype 6, RA56a – RA of unknown serotype, rOmpA – rOmpA induced for 4 h, and M – protein ladder with labelled molecular weights. The positions of the 65-kDa rOmpA and the 42-kDa OmpA proteins are indicated by the arrows
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