1.Research progress in active substances and their mechanisms of action against porcine epidemic diarrhea virus.
Yu LIU ; Sisi SU ; Ziqian WANG ; Jiahao WU ; Hongwei CHEN ; Hongzao YANG
Chinese Journal of Biotechnology 2025;41(7):2519-2533
Porcine epidemic diarrhea virus (PEDV) is an intestinal coronavirus that can cause porcine epidemic diarrhea, leading to diarrhea, vomiting, weight loss, and even death in piglets. Due to the diversity of PEDV strains, traditional vaccines are difficult to sustainably and effectively prevent and control PEDV. This article reviews the strategies and mechanisms of active substances in regulating intracellular signaling pathways, viral proteins, and microbial metabolites to enhance the host immune function against PEDV. It emphasizes the prevention of PEDV resistance and the potential harm of PEDV breaking through interspecies barriers to the human society, aiming to provide reliable theoretical support for the development of new antiviral drugs or vaccines.
Porcine epidemic diarrhea virus/immunology*
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Animals
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Swine
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Swine Diseases/prevention & control*
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Antiviral Agents/pharmacology*
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Coronavirus Infections/virology*
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Viral Vaccines/immunology*
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Humans
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Signal Transduction
2.Development and evaluation of a competitive ELISA based on a porcine neutralizing Fab antibody against Senecavirus A.
Yubin LIANG ; Xueqing MA ; Yixuan HE ; Caihe WANG ; Kun LI ; Pinghua LI ; Yuanfang FU ; Zengjun LU ; Xiaohua DU ; Xia LIU ; Pu SUN
Chinese Journal of Biotechnology 2025;41(7):2748-2759
Senecavirus A (SVA) is a major viral pathogen causing disease in pigs, and effective monitoring of SVA infection is critical for disease control. In this study, we aimed to develop a reliable ELISA method for rapidly detecting neutralizing antibodies against SVA. We used HEK293F cells to express an SVA-specific porcine Fab antibody and verified the biological activity of the Fab antibody by indirect ELISA, immunofluorescence assay, virus neutralization test, and Western blotting. The Fab antibody was biotinylated and used as a competitive antibody to establish a competitive ELISA (C-ELISA) for detecting neutralizing antibodies against SVA. We then evaluated the C-ELISA in terms of sensitivity, specificity, repeatability, and result agreement rate with the VNT. The results showed that we successfully prepared an SVA-specific porcine Fab antibody, which showed high affinity for SVA. We named this antibody 1M33Fab and designated it as Bio-1M33Fab after biotin labeling. The assay conditions were optimized as follows: the coating concentration of SVA particles being 1 μg/mL, the working concentration of Bio-1M33Fab being 0.5 μg/mL, the optimal serum dilution of 1:10, and the optimal dilution of enzyme-labeled avidin being 1:30 000. At a percent inhibition (PI) of 47%, the assay demonstrated the highest sensitivity (96.88%) and specificity (100%), with no cross-reactivity observed with the positive sera of major porcine viral diseases. The intra-assay coefficient of variation ranged from 1.12% to 7.34%, while the inter-assay coefficient of variation ranged from 1.10% to 8.97%, indicating good repeatability. In the detection of 224 clinical pig serum samples, C-ELISA and VNT showed a result agreement rate of 93.75%. In conclusion, we successfully develop a C-ELISA method for detecting neutralizing antibodies against SVA by using a porcine-derived Fab antibody, which lays a foundation for the development of detection kits.
Animals
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Swine
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Antibodies, Neutralizing/immunology*
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Enzyme-Linked Immunosorbent Assay/methods*
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Immunoglobulin Fab Fragments/immunology*
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Antibodies, Viral/immunology*
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Picornaviridae/immunology*
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Humans
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HEK293 Cells
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Swine Diseases/diagnosis*
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Picornaviridae Infections/diagnosis*
3.Nucleocapsid protein from porcine epidemic diarrhea virus isolates can antagonize interferon-λ production by blocking the nuclear factor-κB nuclear translocation.
Ying SHAN ; Zi-Qi LIU ; Guo-Wei LI ; Cong CHEN ; Hao LUO ; Ya-Jie LIU ; Xun-Hui ZHUO ; Xing-Fen SHI ; Wei-Huan FANG ; Xiao-Liang LI
Journal of Zhejiang University. Science. B 2018;19(7):570-580
Porcine epidemic diarrhea virus (PEDV) is a highly infectious pathogen that can cause severe diseases in pigs and result in enormous economic losses in the worldwide swine industry. Previous studies revealed that PEDV exhibits an obvious capacity for modulating interferon (IFN) signaling or expression. The newly discovered type III IFN, which plays a crucial role in antiviral immunity, has strong antiviral activity against PEDV proliferation in IPEC-J2 cells. In this study, we aimed to investigate the effect of PEDV nucleocapsid (N) protein on type III IFN-λ. We found that the N proteins of ten PEDV strains isolated between 2013 and 2017 from different local farms shared high nucleotide identities, while the N protein of the CV777 vaccine strain formed a monophyletic branch in the phylogenetic tree. The N protein of the epidemic strain could antagonize type III IFN, but not type I or type II IFN expression induced by polyinosinic-polycytidylic acid (poly(I:C)) in IPEC-J2 cells. Subsequently, we demonstrated that the inhibition of poly(I:C)-induced IFN-λ3 production by PEDV N protein was dependent on the blocking of nuclear factor-κB (NF-κB) nuclear translocation. These findings might help increase understanding of the pathogenesis of PEDV and its mechanisms for evading the host immune response.
Active Transport, Cell Nucleus
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Animals
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Coronavirus Infections
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immunology
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veterinary
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virology
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Genes, Viral
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Host-Pathogen Interactions
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immunology
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Interferons
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antagonists & inhibitors
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biosynthesis
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genetics
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Interleukins
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antagonists & inhibitors
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biosynthesis
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genetics
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NF-kappa B
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metabolism
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Nucleocapsid Proteins
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genetics
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immunology
;
physiology
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Porcine epidemic diarrhea virus
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genetics
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pathogenicity
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physiology
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Promoter Regions, Genetic
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Swine
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Swine Diseases
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immunology
;
virology
4.Immune Response of Recombinant Pseudorabies Virus rPRV-VP2 Expressing VP2 Gene of Porcine Parvovirus in Mice.
Pengfei FU ; Xinlong PAN ; Qiao HAN ; Xingwu YANG ; Qianlei ZHU ; Xiaoqing GUO ; Yu ZHANG ; Hongying CHEN
Chinese Journal of Virology 2016;32(2):195-202
In order to develop a combined live vaccine that will be used to prevent against porcine parvovirus (PPV) and Pseudorabies virus (PRV) infection, the VP2 gene of PPV was inserted into the transfer vector plasmid pG to produce the recombinant plasmid pGVP2. The plasmid pGVP2 and the genome of PRV HB98 attenuated vaccine were transfected by using lipofectamine into swine testis cells for the homologous recombination. The recombinant virus rPRV-VP2 was purified by selection of green fluorescence plaques for five cycles. 6-week-old female Kunming mice were immunized intramuscularly with attenuated PRV parent HB98 strain, commercial inactivated vaccine against PPV, recombinant virus, DMEM culture solution. The injections were repeated with an equivalent dose after 2 weeks in all of the groups, and then challenged with the virulent PRV NY strain at 7 weeks after the first immunization. The recombinant virus rPRV-VP2 was successfully generated, and the recombinant virus could effectively elicite anti-PPV and PRV antibody and significant cellular immune response as indicated by anti-PPV ELISA and HI, PRV-neutralizing assay and flow cytometry. The challenge assay indicated that recombinant virus could protect the mice against the virulent PRV challenge. These results demonstrated that the recombinant virus can be a candidate recombinant vaccine strain for the prevention of PRV and PPV.
Animals
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Antibodies, Viral
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immunology
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Antigens, Viral
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administration & dosage
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genetics
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immunology
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Capsid Proteins
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administration & dosage
;
genetics
;
immunology
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Female
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Gene Expression
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Genetic Vectors
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genetics
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metabolism
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Herpesvirus 1, Suid
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genetics
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metabolism
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Mice
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Parvovirus, Porcine
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genetics
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immunology
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Swine
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Swine Diseases
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immunology
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prevention & control
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virology
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Viral Vaccines
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administration & dosage
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genetics
;
immunology
5.Assessing PCV2 antibodies in field pigs vaccinated with different porcine circovirus 2 vaccines using two commercial ELISA systems.
Min Kyoung SHIN ; Seung Hyun YOON ; Myung Hwui KIM ; Young Soo LYOO ; Seung Won SUH ; Han Sang YOO
Journal of Veterinary Science 2015;16(1):25-29
Porcine circovirus type 2 (PCV2) is the primary causative agent for post-weaning, multisystemic, wasting syndrome. Consequently, serologic detection of and vaccination against PCV2 are important for the swine industry. Among several serological tests, the enzyme-linked immunosorbent assay (ELISA) is commonly used to measure anti-PCV2 antibody levels. In the present study, we used two commercial ELISA systems to comparatively evaluate anti-PCV2 antibodies in field pigs treated with three different PCV2 vaccines. Among a total of 517 serum samples, the results of the two ELISAs were fully concordant for 365 positive and 42 negative samples, indicating 78.7% agreement. In addition, the Pearson coefficient (0.636) indicated a moderate correlation between data from the two ELISAs. Results from the farms with pigs vaccinated with the three different PCV2 vaccines demonstrated that most of the vaccinated animals underwent seroconversion. However, the increase and duration of antibody titers varied depending on the vaccine, the presence of maternal antibodies, and the vaccination program. PCV2 serologic status and anti-PCV2 antibody levels of herds from this study could be utilized to determine the best timing for vaccination and assessing vaccination compliance.
Aging
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Animals
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Antibodies, Viral/*blood
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Circovirus/*classification/immunology
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Enzyme-Linked Immunosorbent Assay/methods/*veterinary
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Female
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Porcine Postweaning Multisystemic Wasting Syndrome/blood/immunology/*prevention & control
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Republic of Korea/epidemiology
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Swine
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Swine Diseases/*prevention & control/virology
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Viral Vaccines/*immunology
6.Construction and identification of a recombinant PRRSV expressing ORF2 of porcine circovirus type 2.
Tingjie ZHANG ; Xing LIU ; Tao SUN ; Xuejiao ZHU ; Baochao FAN ; Juan BAI ; Ping JIANG
Chinese Journal of Virology 2015;31(1):65-73
Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are very two important pathogens that have coursed huge economic losses in swine production in worldwide. In this study,a vector pCMV-TJM containing the full-length cDNA clone of PRRSV attenuated strain TJM-F92 was firstly constructed by PCR method. Then a gene sequence containing Afl II/Mlu I e restriction enzyme sites and a transcription regulatory sequence for ORF6 (TRS6) was inserted be- tween ORF7 and 3'UTR, yielding a expression vector pCMV-TJM-TRS. Subsequently, a plasmid pCMV-TJM-Cap was constructed by cloning of PCV2 ORF2 gene into the unique sites Afl II /Mlu I of pCMV- TJM-TRS plasmid DNA. Then three recombinant PRRSV, rTJM, rTJM/TRS and rTJM/Cap, were rescued by transfection of pCMV-TJM, pCMV-TJM-TRS and pCMV-TJM-Cap into Marc-145 cells, respectively,and confirmed by the genome sequence, restriction enzyme digestion, Western Blot and IFA. They all had the molecular markers which was different from the parent virus. The growth characteristics of the rescued viruses were similar to that of parent virus. rTJM/Cap could also express efficiently PCV2 Cap protein in Marc-145 cells. At passage 8, it still had PCV2 ORF2 gene which examined by RT-PCR. It indicated that the full-length cDNA clone of PRRSV attenuated strain TJM-F92 and recombinant PRRSV rTJM/Cap expressing PCV2 Cap protein were successfully constructed. It made an important foundation for studying on the pathogenic mechanisms of PRRSV and PRRSV-PCV2 vaccine in the future.
Animals
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Capsid Proteins
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genetics
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immunology
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Cell Line
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Circoviridae Infections
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veterinary
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virology
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Circovirus
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classification
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genetics
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metabolism
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Gene Expression
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Open Reading Frames
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Porcine Reproductive and Respiratory Syndrome
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virology
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Porcine respiratory and reproductive syndrome virus
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genetics
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metabolism
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Recombination, Genetic
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Swine
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Swine Diseases
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virology
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Viral Vaccines
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genetics
;
immunology
7.Seroprevalence of Trichinella sp. in Wild Boars (Sus scrofa) from Yanggu-gun, Gangwon-do, Korea.
Hye Jung LEE ; Ok Sik CHUNG ; Jae Lip KIM ; Seung Ha LEE ; Young Bok YOO ; Min SEO
The Korean Journal of Parasitology 2015;53(2):233-236
A total 7 outbreaks of trichinellosis have occurred in Korea, mostly as a result of consumption of raw wild boar (Sus scrofa) meat. Since only 1 serological survey on wild boars had yet been performed in Korea, the present study aimed to estimate the prevalence of trichinellosis in wild boars and some species of rodents by artificial digestion and serological examinations in Yanggu-gun, Gangwon-do, the endemic area of trichinellosis. Both the wild boar and rodent muscle samples revealed no Trichinella larvae by direct examination and artificial digestion method. However, serological examinations revealed that 4 wild boar sera samples out of 118 (3.4%) were positive to Trichinella antigen. Although the recovery of Trichinella larvae ended in a failure, it is proved for the first time that the sylvatic cycle of Trichinella has been maintained in wild boars of Gangwon-do, Korea.
Animals
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Antibodies, Helminth/*blood
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Antigens, Helminth/blood
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Female
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Male
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Republic of Korea/epidemiology
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Seroepidemiologic Studies
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Sus scrofa
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Swine
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Swine Diseases/*blood/diagnosis/epidemiology/parasitology
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Trichinella/classification/genetics/immunology/*isolation & purification
8.Protective efficacy of a high-growth reassortant swine H3N2 inactivated vaccine constructed by reverse genetic manipulation.
Feng WEN ; Ji Hong MA ; Hai YU ; Fu Ru YANG ; Meng HUANG ; Yan Jun ZHOU ; Ze Jun LI ; Guang Zhi TONG
Journal of Veterinary Science 2014;15(3):381-388
Novel reassortant H3N2 swine influenza viruses (SwIV) with the matrix gene from the 2009 H1N1 pandemic virus have been isolated in many countries as well as during outbreaks in multiple states in the United States, indicating that H3N2 SwIV might be a potential threat to public health. Since southern China is the world's largest producer of pigs, efficient vaccines should be developed to prevent pigs from acquiring H3N2 subtype SwIV infections, and thus limit the possibility of SwIV infection at agricultural fairs. In this study, a high-growth reassortant virus (GD/PR8) was generated by plasmid-based reverse genetics and tested as a candidate inactivated vaccine. The protective efficacy of this vaccine was evaluated in mice by challenging them with another H3N2 SwIV isolate [A/Swine/Heilongjiang/1/05 (H3N2) (HLJ/05)]. Prime and booster inoculation with GD/PR8 vaccine yielded high-titer serum hemagglutination inhibiting antibodies and IgG antibodies. Complete protection of mice against H3N2 SwIV was observed, with significantly reduced lung lesion and viral loads in vaccine-inoculated mice relative to mock-vaccinated controls. These results suggest that the GD/PR8 vaccine may serve as a promising candidate for rapid intervention of H3N2 SwIV outbreaks in China.
Animals
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Female
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Influenza A Virus, H3N2 Subtype/*genetics/immunology
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Influenza Vaccines/genetics/immunology/*therapeutic use
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Mice
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Mice, Inbred BALB C
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Orthomyxoviridae Infections/immunology/*prevention & control/virology
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Reassortant Viruses/genetics/immunology
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Reverse Genetics/methods/*veterinary
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Swine
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Swine Diseases/immunology/*prevention & control/virology
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Vaccines, Inactivated
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Virus Replication
9.Dietary germanium biotite supplementation enhances the induction of antibody responses to foot-and-mouth disease virus vaccine in pigs.
Jin A LEE ; Bock Gie JUNG ; Myunghwan JUNG ; Tae Hoon KIM ; Han Sang YOO ; Bong Joo LEE
Journal of Veterinary Science 2014;15(3):443-447
We evaluated the potential ability of germanium biotite (GB) to stimulate the production of antibodies specific for foot-and-mouth disease virus (FMDV). To this aim, we measured the total FMDV-specific antibody responses and IgM production after vaccination against FMD both experimentally and in the field. GB supplementation with FMDV vaccination stimulated the production of anti-FMDV antibodies, and effectively increased IFN-gamma and TNF-alpha levels. These results suggest that GB may be a novel alternative feed supplement that can serve as a boosting agent and an immunostimulator for increasing the efficacy of FMDV vaccination in pigs.
Adjuvants, Immunologic/therapeutic use
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Aluminum Silicates/*therapeutic use
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Animals
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Antibodies, Viral/*immunology
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Antibody Formation/drug effects
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*Dietary Supplements
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Ferrous Compounds/*therapeutic use
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Foot-and-Mouth Disease/*immunology/prevention & control
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Foot-and-Mouth Disease Virus/immunology
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Germanium/*therapeutic use
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Swine
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Swine Diseases/immunology/prevention & control/*virology
10.Induction of antibody and interferon-gamma production in mice immunized with virus-like particles of swine hepatitis E virus.
Young Jo SONG ; Woo Jung PARK ; Seul Kee LEE ; Joong Bok LEE ; Seung Yong PARK ; Chang Seon SONG ; Sang Won LEE ; Kun Ho SEO ; Young Sun KANG ; Jae Young SONG ; In Soo CHOI
Journal of Veterinary Science 2014;15(4):575-578
Virus-like particles (VLPs) composed of the truncated capsid protein of swine hepatitis E virus (HEV) were developed and immune responses of mice immunized with the VLPs were evaluated. IgG titers specific for the capsid protein of swine HEV were significantly higher for all groups of mice immunized with the VLPs than those of the negative control mice. Splenocytes from mice immunized with the VLPs also produced significantly greater quantities of interferon (IFN)-gamma than interleukin (IL)-4 and IL-10. These newly developed swine HEV VLPs have the capacity to induce antigen-specific antibody and IFN-gamma production in immunized mice.
Animals
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Antibodies, Viral/blood
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Capsid Proteins/immunology
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Female
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Hepatitis E/immunology/*veterinary/virology
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Hepatitis E virus/*immunology
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Immunization/*veterinary
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Interferon-gamma/blood
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Mice
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Mice, Inbred BALB C
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Swine
;
Swine Diseases/*immunology/virology
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Vaccines, Virus-Like Particle/immunology
;
Viral Hepatitis Vaccines/*immunology

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