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1 2 Nakane, T. (2014). Recognition of the different structural forms of the capsid protein determines the outcome following infection with Porcine Circovirus Type 2 All about SWINE 44, PCV2 porcine circovirus type PMWS porcine multi-systemic wasting syndrome 1990 PCV PCV2 PCVAD porcine circovirus associated disease PCVAD PCV2 PCVAD PCVAD B.R. Trible R.R.R. Rowland B.R. Trible PCV2 Trible PCV2 5 1 PCV2 PCVAD PCV2 CP capsid protein 16

2 1 PCV2 5 PCV2 5 PCV 1970 PK-15 Tischer 1974 DNA PCV PCV Tischer 1982 Tischer PCV PMWS Clark 1997 Harding 1997 DNA PK-15 PCV 70 Meehan 1998 PK-15 PCV1 PMWS PCV2 PCV2 PCV2a GenBank accession #AF PCV2b GenBank accession #AF055394; Mehann PCV2c GenBank accession #EU PCV2a PCV2b 17

PCV2b PCVAD Segales 2008 2005 Horlen 2007 PCV2a PCV2b PCV2a 2b Opriessnig 2008 GenBank PCV2 139 3 3 2.

3 PCV2b PCVAD Segales Horlen 2007 PCV2a PCV2b PCV2a 2b Opriessnig 2008 GenBank PCV GenBank PCV PCV2 DNA PCV1 1,759 PCV2a PCV2b PCV2c 1,768 1,767 1,767 PCV2 PCV PCV2a PCV2b PCV2c 95 Fenaux ORF open reading frames ORF1 DNA Rept Rep Mankertz Hillenbrand 2001 ORF CP 3 ORF3 ORF1 Liu 2005 PCV2 ORF3 18

Chaiyakul 2010 3. PCV2 5 PCV 60 CP PCV2 VLPs virus-like particle Khaya 2011 CP CP Liu 2001 Misinzo 2006 Shuai 2008 PCV2 a CP CP b CP 60 CP VLPs 2011 Khayat UCSF Chimera PDB ID 3R0R; Pettersen 2004 4.

4 Chaiyakul PCV2 5 PCV 60 CP PCV2 VLPs virus-like particle Khaya 2011 CP CP Liu 2001 Misinzo 2006 Shuai 2008 PCV2 a CP CP b CP 60 CP VLPs 2011 Khayat UCSF Chimera PDB ID 3R0R; Pettersen PCV2 CP 3 PCVAD PMWS PDNS porcine dermatitis and nephropathy syndromeape acute pulmonary edema 50Segales 2012 PCV2 PCV2 Meehan 1998 Rossell 2000 Wellenberg 2004 PCV2 PCVAD PRRSV SIV PPV Hps M.hyo APP PCVAD PRRSV PCV2 PRRSV 19

5 PCV2 PCV2 PCV2 PRRSV Allan 2000 Rovira 2002 Sinha 2011 Trible 2012 PCV2 Darwich 2004 PCV2 PCVAD PCV2a ORF2 4 1 PCV CP PCV PCV1 PCV2 ORF2 VLP PCV2a 1ml 2ml PCV2 PCV2a PCV2b PCVAD 3 7 Elisa VLP 6 VLP VLP VLP 20

6 VLP VLP VLP Capsid 2 2 Capsid PCV PCV PCV PK cpcv1-2 Spodoptera frugiperda 2 2 BacPCV2- Orf2;98-99 PK Spodoptera frugiperda 2 2 N W ml 1 2ml ml mL CP Mahe 2000 Lekcharoensuk 2004 Mahe CP 12 PCV2b CP Lekcharoensuk PCV1 PCV2a ORF2 PCV DNA 7 CP Trible 2011 A-D PCV2 PCV2 CP PCV2a PCV2b 21

7 4 A-D 5. PCV2 CP 1 Trible 2011 CP PCV2 PCa PCV2 VX PMWS PM PCVAD PD CP 50 CP PCV2 PCVAD CP C 2. PCV2 1 PCVAD C C 15 C PEPSCAN CP Y 173- Tyr F 174- Phe Q 175- Gln K 179- Lys GenBank 462 PCV2 22

8 Table 2 Summary of antibody responses to PCV2 capsid polypeptides. Name Epitope Regions PC a Vx PM PD b a Key: PC = PCV2-infected; Vx = vaccinated; PM = PMWS; PD =PDNS. b Relative binding activity. Key: (-) no measureable binding activity; (+) low binding activity; (++) intermediate binding activity; (+++) high binding activity ; (++++) very high binding activity Clin Vaccine Immuol.18: C PEPSCAN 5 CP 4 CP A CP 173-Tyr 174-Phe 175-Gln B CP VLPs C CP VLPs VLPs 3 CP 173-Tyr VLPs CP CP CP VLP 4 Trible 2012 CP VLPs CP PCV2 RRSV VLPs VLPs CP Bac-CP Khayat 2011 CP VLPs 23

Ub Ub-CP PCV2 IFA 320 PCV2 26 3 8 3 7 Bac-CP Intervet- Schering Plough, hayat 2011 Ub- CP Ub 4 7 2ml Ub-CP Ub-CP 43-233 2ml MEM 50 /ml Emulsigen-D 20 vol/vol;

9 Ub Ub-CP PCV2 IFA 320 PCV Bac-CP Intervet- Schering Plough, hayat 2011 Ub- CP Ub 4 7 2ml Ub-CP Ub-CP ml MEM 50 /ml Emulsigen-D 20 vol/vol; MVP Technologies 30 g/ml 0.01 Non- IM Bac-CP Ub-CP PCV2b PRRSV 10 5 TCID50 1.5ml 3ml PCV PRRS 2 PRRSV PCV2 PCV2 Ub-CP 8 6 Non-IM 8 4 Bac-CP

10 Group Non-IM Bac-CP Ub-CP Control n Immunogen None Bac-CP vaccine Ub-CP None PCV2/PRRSV Yes Yes Yes No Trible B R et al. J. Virol. 2012;86: Non-IM Bac-CP ; (Intervet-Schering Plough) Ub-CP ; Ub Control ; PCV2 PCV2 IFA Ub-CP Bac-CP PCV2 Non-IM IFA Contorol 2 PCV2 9A. PCV2 4 3 Non-IM 21 Ub-CP 9B. PCV2 CP PCV PCV2 35 Bac-CP Non-IM Ub-CP Non-IM Ub-CP 10. PCV2 4 3 CP CP Ub-CP Bac-CP 2 Contorol PCV2 PCV2 PCR PRRSV PCV2 Ub-CP Non-Im 14 PCV2 DNA 25

11 Bac-CP 11. PCV2 4 3 VLP CP Bac-CP PCV2 CP CP CP PCV2 CP Ub-CP CP PCV2 Non- 26

IM PCV2 PCVAD CP 169-180 12 VLP CP 3 PCV2 3 PCVAD PCVAD CP 169-180

12 IM PCV2 PCVAD CP VLP CP 3 PCV2 3 PCVAD PCVAD CP PCVAD PCVAD PCVAD 13. PCV Benjamin R. Trible, Maureen Kerrigan, Nicholas Crossland, Megan Potter, Kay Faaberg, Richard Hesse, and Raymond R. R. Rowland Antibody Recognition of Porcine Circovi- 27

13 rus Type 2 Capsid Protein Epitopes after Vaccination, Infection, and Disease. Clin Vaccine Immuol. 18: Benjamin R. Trible a, Alejandro Ramirezb, Andrew Sudditha, Alexandra Fuller a, Maureen Kerrigana, Richard Hessea, Jerome Nietfelda, Baoqing Guoc, Eileen Thackerd, Raymond R.R. Rowlanda Antibody responses following vaccination versus infection in a porcine circovirus-type 2 (PCV2) disease model show distinct differences in virus neutralization and epitope recognition. Vaccine 30: Benjamin R. Trible, Raymond R.R. Rowland Genetic variation of porcine circovirus type 2 (PCV2) and its relevance to vaccination, pathogenesis and diagnosis. Virus Res. 164: Benjamin R. Trible, Andrew W. Suddith, Maureen A. Kerrigan, Ada G. Cino-Ozuna, Richard A. Hesse and Raymond R. R. Rowland Recognition of the Different Structural Forms of the Capsid Protein Determines the Outcome following Infection with Porcine Circovirus Type 2. J. Virol. 86: Benjamin R. Trible, R. R. Rowland Protective Immunity vs. Immunopathogenesis: Recognition of the structural form of the PCV2 capsid determines the outcome. 6 Elisa Crisci a, Juan Barcenab, Maria Montoyaa Virus-like particle-based vaccines for animal viral infections. Inmunología, 32: :