PMID- 38005998
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20231127
IS  - 2076-393X (Print)
IS  - 2076-393X (Electronic)
IS  - 2076-393X (Linking)
VI  - 11
IP  - 11
DP  - 2023 Oct 31
TI  - Development of Polycistronic Baculovirus Surface Display Vectors to 
      Simultaneously Express Viral Proteins of Porcine Reproductive and Respiratory 
      Syndrome and Analysis of Their Immunogenicity in Swine.
LID - 10.3390/vaccines11111666 [doi]
LID - 1666
AB  - To simultaneously express and improve expression levels of multiple viral 
      proteins of a porcine reproductive and respiratory syndrome virus (PRRSV), 
      polycistronic baculovirus surface display vectors were constructed and 
      characterized. We engineered polycistronic baculovirus surface display vectors, 
      namely, pBacDual Display EGFP(BacDD)-2GP2-2GP4 and pBacDD-4GP5N34A/N51A (mtGP5), 
      which simultaneously express and display the ectodomain of His-tagged 
      GP2-gp64TM-CTD, His-tagged GP4-gp64TM-CTD, and His-tagged mtGP5-gp64TM-CTD fusion 
      proteins of PRRSV on cell membrane of Sf-9 cells. Specific pathogen-free (SPF) 
      pigs were administered intramuscularly in 2 doses at 21 and 35 days of age with 
      genetic recombinant baculoviruses-infected cells. Our results revealed a high 
      level of ELISA-specific antibodies, neutralizing antibodies, IL-4, and IFN-gamma in 
      SPF pigs immunized with the developed PRRSV subunit vaccine. To further assess 
      the co-expression efficiency of different gene combinations, pBacDD-GP2-GP3-2GP4 
      and pBacDD-2mtGP5-2M constructs were designed for the co-expression of the 
      ectodomain of His-tagged GP2-gp64TM-CTD, His-tagged GP3-gp64TM-CTD, and 
      His-tagged GP4-gp64TM-CTD proteins as well as the ectodomain of His-tagged 
      mtGP5-gp64TM-CTD and His-tagged M-gp64TM-CTD fusion proteins of PRRSV. To develop 
      an ELISA assay for detecting antibodies against PRRSV proteins, the sequences 
      encoding the ectodomain of the GP2, GP3, GP4, mtGP5, and M of PRRSV were 
      amplified and subcloned into the pET32a vector and expressed in E. coli. In this 
      work, the optimum conditions for expressing PRRSV proteins were evaluated, and 
      the results suggested that 4 x 10(5) of Sf-9 cells supplemented with 7% fetal 
      bovine serum and infected with the recombinant baculoviruses at an MOI of 20 for 
      three days showed a higher expression levels of the protein. Taken together, the 
      polycistronic baculovirus surface display system is a useful tool to increase 
      expression levels of viral proteins and to simultaneously express multiple viral 
      proteins of PRRSV for the preparation of subunit vaccines.
FAU - Hsu, Chao-Yu
AU  - Hsu CY
AD  - Department of Medical Research, Tungs' Taichung Metroharbor Hospital, Taichung 
      435, Taiwan.
AD  - Ph.D. Program in Translational Medicine, National Chung Hsing University, 
      Taichung 402, Taiwan.
FAU - Jang, Yun
AU  - Jang Y
AD  - Institute of Molecular Biology, National Chung Hsing University, Taichung 402, 
      Taiwan.
FAU - Huang, Wei-Ru
AU  - Huang WR
AD  - Institute of Molecular Biology, National Chung Hsing University, Taichung 402, 
      Taiwan.
AD  - The iEGG and Animal Biotechnology Center, National Chung Hsing University, 
      Taichung 402, Taiwan.
FAU - Wang, Chi-Young
AU  - Wang CY
AUID- ORCID: 0000-0001-5407-3414
AD  - Department of Veterinary Medicine, National Chung Hsing University, Taichung 402, 
      Taiwan.
FAU - Wen, Hsiao-Wei
AU  - Wen HW
AUID- ORCID: 0000-0003-3427-1768
AD  - Department of Food Science and Biotechnology, National Chung Hsing University, 
      Taichung 402, Taiwan.
FAU - Tsai, Pei-Chien
AU  - Tsai PC
AD  - Department of Life Sciences, National Chung Hsing University, Taichung 402, 
      Taiwan.
FAU - Yang, Cheng-Yao
AU  - Yang CY
AUID- ORCID: 0000-0002-8694-9561
AD  - Graduate Institute of Veterinary Pathobiology, National Chung Hsing University, 
      Taichung 402, Taiwan.
FAU - Munir, Muhammad
AU  - Munir M
AUID- ORCID: 0000-0003-4038-0370
AD  - Division of Biomedical and Life Sciences, Faculty of Health and Medicine, 
      Lancaster University, Lancaster LA1 4YW, UK.
FAU - Liu, Hung-Jen
AU  - Liu HJ
AUID- ORCID: 0000-0002-1460-1494
AD  - Ph.D. Program in Translational Medicine, National Chung Hsing University, 
      Taichung 402, Taiwan.
AD  - Institute of Molecular Biology, National Chung Hsing University, Taichung 402, 
      Taiwan.
AD  - The iEGG and Animal Biotechnology Center, National Chung Hsing University, 
      Taichung 402, Taiwan.
AD  - Department of Life Sciences, National Chung Hsing University, Taichung 402, 
      Taiwan.
AD  - Rong Hsing Research Center for Translational Medicine, National Chung Hsing 
      University, Taichung 402, Taiwan.
LA  - eng
GR  - 109-2313-B-005-006-MY3& 112-2313-B-005-050-MY3/Ministry of Science and Technology 
      of Taiwan/
GR  - 112S0023A/The Ministry of Education (MOE) in Taiwan/
PT  - Journal Article
DEP - 20231031
PL  - Switzerland
TA  - Vaccines (Basel)
JT  - Vaccines
JID - 101629355
PMC - PMC10674950
OTO - NOTNLM
OT  - IFN-gamma
OT  - IL-4
OT  - baculovirus surface display vectors
OT  - neutralizing antibody
OT  - porcine reproductive and respiratory syndrome virus
COIS- We declare that we have no competing interests.
EDAT- 2023/11/25 12:46
MHDA- 2023/11/25 12:47
PMCR- 2023/10/31
CRDT- 2023/11/25 01:31
PHST- 2023/10/01 00:00 [received]
PHST- 2023/10/21 00:00 [revised]
PHST- 2023/10/30 00:00 [accepted]
PHST- 2023/11/25 12:47 [medline]
PHST- 2023/11/25 12:46 [pubmed]
PHST- 2023/11/25 01:31 [entrez]
PHST- 2023/10/31 00:00 [pmc-release]
AID - vaccines11111666 [pii]
AID - vaccines-11-01666 [pii]
AID - 10.3390/vaccines11111666 [doi]
PST - epublish
SO  - Vaccines (Basel). 2023 Oct 31;11(11):1666. doi: 10.3390/vaccines11111666.