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1.
Adv Mater ; 36(21): e2309189, 2024 May.
Article in English | MEDLINE | ID: mdl-38530975

ABSTRACT

Surface plasmons in 2D materials such as graphene exhibit exceptional field confinement. However, the low electron density of majority of 2D materials, which are semiconductors or semimetals, has limited their plasmons to mid-wave or long-wave infrared regime. This study demonstrates that a 2D Ti3C2Tx MXene with high electron density can not only support strong plasmon confinement with an acoustic plasmon mode in the short-wave infrared region, but also provide ultrahigh nonlinear responses. The acoustic MXene plasmons (AMPs) in the MXene (Ti3C2Tx)-insulator (SiO2)-metal (Au) nanostructure generate in the 1.5-6.0 µm wavelength range, exhibiting a two orders of magnitude reduction in wavelength compared to wavelength in free space. Furthermore, AMP resonators with patterned Au rods exhibit a record-high nonlinear absorption coefficient of 1.37 × 10-2 m W-1 at wavelength of 1.56 µm, ≈3 orders of magnitude greater than the highest value recorded for other 2D materials. These results indicate that MXenes can overcome fundamental plasmon wavelength limitations of previously studied 2D materials, providing groundbreaking opportunities in nonlinear optical applications, including all-optical processing and ultrafast optical switching.

2.
Zygote ; 31(6): 577-581, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37905414

ABSTRACT

Klotho is a protein that plays different functions in female fertility. We have previously reported that klotho protein supplementation during in vitro maturation improves porcine embryo development, while klotho knockout for somatic cell cloning completely blocks full-term pregnancy in vivo. However, the effects of the microinjection of klotho protein or klotho knockdown dual vector in porcine embryos at different time points and the specific molecular mechanisms remain largely unknown. In this study, we injected the preassembled cas9 + sgRNA dual vector, for klotho knockdown, into the cytoplasm of the germinal vesicle stage of oocytes and into porcine embryos after 6-h parthenogenetic activation. Similarly, the klotho protein was inserted into the cytoplasm of germinal vesicle stage oocytes and porcine embryos after 6-h parthenogenetic activation. Compared with the controls, the microinjection of klotho dual vector markedly decreased the blastocyst formation rates in germinal vesicle stage oocytes and activated embryos. However, the efficiency of blastocyst formation when klotho protein was inserted before in vitro maturation was significantly higher than that after klotho protein insertion into parthenogenetically activated embryos. These results indicated that klotho knockdown may impair embryo development into blastocyst irrespective of injection timing. In addition, klotho protein injection timing in pig embryos may be an important factor for regulating embryo development.


Subject(s)
Oocytes , RNA, Guide, CRISPR-Cas Systems , Pregnancy , Animals , Female , Swine , Oocytes/physiology , Blastocyst , Embryonic Development/genetics , Parthenogenesis
3.
J Gen Virol ; 104(8)2023 08.
Article in English | MEDLINE | ID: mdl-37650730

ABSTRACT

Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically important virus within the swine industry. The virus causes respiratory disease and reproductive failure. Two species of PRRSV-I and II are co-dominant, yet no effective vaccination strategy has been developed to protect against these two types. With an aim to develop a chimeric vaccine strain to protect against both types, in this study, a chimeric porcine reproductive and respiratory syndrome virus (PRRSV) type I and II was rescued using reverse genetics for the first time. Four chimeric infectious clones were designed based on the genomic arrangement of the structural proteins. However, only the clone carrying the transcriptional regulatory sequence (TRS) and ORF6 of a PRRSV-I and ORF6 of a PRRSV-II generated a viable recombinant virus, suggesting that concurrent expression of ORF6 from both parental viruses is essential for the recovery of type I and II chimeric PRRSV. The chimeric virus showed significantly lower replication ability than its parental strains in vitro, which was improved by serial passaging. In vivo, groups of pigs were inoculated with either the chimeric virus, one of the parental strains, or PBS. The chimeric virus replicated in pig tissue and was detected in serum 7 days post-inoculation. Serum neutralization tests indicated that pigs inoculated with the chimeric virus elicited neutralizing antibodies that inhibited infection with strains of both species and with greater coverage than the parental viruses. In conclusion, the application of this technique to construct a chimeric PRRSV holds promise for the development of a highly effective modified live vaccine candidate. This is particularly significant since there are currently no approved commercial divalent vaccines available to combat PRRSV-I and II co-infections.


Subject(s)
Coinfection , Porcine respiratory and reproductive syndrome virus , Animals , Swine , Antibodies, Neutralizing , Porcine respiratory and reproductive syndrome virus/genetics , Vaccination , Vaccines, Attenuated/genetics
4.
Front Microbiol ; 14: 1202993, 2023.
Article in English | MEDLINE | ID: mdl-37520371

ABSTRACT

High-mobility group box 1 (HMGB1) is a protein that binds to DNA and participates in various cellular processes, including DNA repair, transcription, and inflammation. It is also associated with cancer progression and therapeutic resistance. Despite its known role in promoting tumor growth and immune evasion in the tumor microenvironment, the contribution of HMGB1 to the development of Kaposi's sarcoma (KS) is not well understood. We investigated the effect of HMGB1 on KS pathogenesis using immortalized human endothelial cells infected with Kaposi's sarcoma-associated human herpes virus (KSHV). Our results showed that a higher amount of HMGB1 was detected in the supernatant of KSHV-infected cells compared to that of mock-infected cells, indicating that KSHV infection induced the secretion of HMGB1 in human endothelial cells. By generating HMGB1 knockout clones from immortalized human endothelial cells using CRISPR/Cas9, we elucidated the role of HMGB1 in KSHV-infected endothelial cells. Our findings indicate that the absence of HMGB1 did not induce lytic replication in KSHV-infected cells, but the cell viability of KSHV-infected cells was decreased in both 2D and 3D cultures. Through the antibody array for cytokines and growth factors, CXCL5, PDGF-AA, G-CSF, Emmprin, IL-17A, and VEGF were found to be suppressed in HMGB1 KO KSHV-infected cells compared to the KSHV-infected wild-type control. Mechanistically, phosphorylation of p38 would be associated with transcriptional regulation of CXCL5, PDGF-A and VEGF. These observations suggest that HMGB1 may play a critical role in KS pathogenesis by regulating cytokine and growth factor secretion and emphasize its potential as a therapeutic target for KS by modulating the tumor microenvironment.

5.
J Med Virol ; 95(8): e28978, 2023 08.
Article in English | MEDLINE | ID: mdl-37515534

ABSTRACT

Dengue virus, which belongs to the Flaviviridae family, can induce a range of symptoms from mild to severe, including dengue fever, dengue hemorrhagic fever, and dengue shock syndrome. While infectious cloning technology is a useful tool for understanding viral pathogenesis and symptoms, it exhibits limitations when constructing the entire Flavivirus genome. The instability and toxicity of the genome to bacteria make its full-length construction in bacterial vectors a time-consuming and laborious process. To address these challenges, we employed the modified infectious subgenomic amplicon (ISA) method in this study, which can potentially be a superior tool for reverse genetic studies on the dengue virus. Using ISA, we generated recombinant dengue viruses de novo and validated their robust replication in both human and insect cell lines, which was comparable to that of the original strains. Moreover, the efficiency of ISA in genetically modifying the dengue virus was elucidated by successfully inserting the gene for green fluorescence protein into the genome of dengue virus serotype 4. Overall, this study highlighted the effectiveness of ISA for genetically engineering the dengue virus and provided a technical basis for a convenient reverse genetics system that could expedite investigations into the dengue virus.


Subject(s)
Dengue Virus , Dengue , Flaviviridae , Flavivirus , Humans , Dengue Virus/genetics , Reverse Genetics/methods , Flavivirus/genetics , Flaviviridae/genetics , Virus Replication/genetics
6.
Virology ; 579: 119-127, 2023 02.
Article in English | MEDLINE | ID: mdl-36669328

ABSTRACT

Codon pair deoptimization (CPD) attenuated type I porcine reproductive and respiratory syndrome virus (PRRSV). Infectious clones covering the full genome of a Korean type I PRRSV (E38) were synthesized, and CPD induced nine synonymous mutants of NSP1 (n = 1) and ORF7 (n = 8). In a trial to rescue live viruses from infectious clones, only four clones with mutations at nt 177 downstream of ORF7 were rescued, which showed a substantial decrease in cellular replication ability. The rescue-failed clones had two common mutation sites with a high minimum free energy and significantly modified RNA secondary structure relative to the original virus. In infected pigs, CPD viruses demonstrated significantly lower replication ability and pathogenicity than the original virus. However, immune response level induced by the attenuated viruses and the original virus was similar. This is the first study to demonstrate that type I PRRSV virulence can be attenuated through CPD application to ORF7.


Subject(s)
Porcine Reproductive and Respiratory Syndrome , Porcine respiratory and reproductive syndrome virus , Viral Vaccines , Viruses , Animals , Swine , Porcine respiratory and reproductive syndrome virus/genetics , Virus Replication/genetics , Codon , Mutation , Viruses/genetics , Immunity , Porcine Reproductive and Respiratory Syndrome/genetics , Viral Vaccines/genetics
7.
J Med Virol ; 95(1)2023 01.
Article in English | MEDLINE | ID: mdl-35869037

ABSTRACT

Many cytokines produced by Kaposi's sarcoma-associated herpesvirus (KSHV)-infected cells have been shown to participate in the pathogenesis of KSHV. Determination of the exact role of cytokines in Kaposi's sarcoma (KS) pathogenesis is limited, however, by the difficulty to manipulate the target genes in human endothelial cells. In this study, we sought to elucidate the role of cytokines in KSHV-infected human immortalized endothelial cell line (HuARLT cells) by knockout (KO) of the corresponding target genes using the CRISPR/Cas9 system. The cytokine production profile of KSHV-infected HuARLT cells was analyzed using a protein array, and several cytokines were found to be highly upregulated following KSHV infection. This study focused on CXCL1, which was investigated by knocked out in HuARLT cells. KSHV-infected CXCL1 KO cells underwent increased cell death compared to KSHV-infected wild-type (WT) cells and mock-infected CXCL1 KO cells. Lytic replication was not observed in KSHV-infected WT nor CXCL1 KO cells. Phosphorylation of STAT3 was significantly suppressed in KSHV-infected CXCL1 KO cells. Additionally, inhibitors of STAT3 and CXCL1 induced cell death in KSHV-infected endothelial cells. Our results show that CXCL1 production is required for the survival of KSHV-infected endothelial cells, and the CXCL1 to STAT3 phosphorylation signaling pathway may be a therapeutic target for KS.


Subject(s)
Herpesvirus 8, Human , Sarcoma, Kaposi , Humans , Herpesvirus 8, Human/physiology , Endothelial Cells , Phosphorylation , Cytokines/metabolism , Chemokine CXCL1/genetics , Chemokine CXCL1/metabolism , STAT3 Transcription Factor/genetics , STAT3 Transcription Factor/metabolism
8.
Pathogens ; 10(12)2021 Dec 20.
Article in English | MEDLINE | ID: mdl-34959599

ABSTRACT

In recent years, porcine circovirus type 2d (PCV2d) has achieved a dominant position worldwide. Various PCV2d capsid-based vaccines have been used to alleviate concerns regarding the emergence of the variant. This study aimed to determine the dosage of recombinant PCV2d capsid protein to induce protective efficacy against experimental challenge with a virulent PCV2d strain. Conventional 3-week-old pigs were intramuscularly inoculated with different doses of the protein (60, 20, 10 and 2 µg). Four weeks after vaccination, all pigs were challenged with pathogenic PCV2d (SNU140003), which was isolated from a farm severely experiencing PCV2-associated disease in Korea. Vaccination with greater than 10 µg of the capsid protein caused a significant (p < 0.05) reduction in PCV2d viremia, lymphoid lesions and lymphoid PCV2 antigen levels in vaccinated challenged pigs compared to unvaccinated challenged pigs. The vaccination also resulted in significantly higher (p < 0.05) titers of neutralizing antibodies against PCV2d. However, the pigs vaccinated with 2 µg had significantly lower neutralizing antibody titers than the other vaccinated groups. They showed a similar level of challenged PCV2d in serum and lymphoid lesion score compared to unvaccinated challenged pigs. The difference in efficacy among the vaccinated groups indicates that there may be a baseline dosage to induce sufficient neutralizing antibodies to prevent viral replication in pigs. In conclusion, at least 10 µg dosage of capsid protein is essential for stable protective efficacy against PCV2d in a pig model.

9.
J Virol ; 95(16): e0079921, 2021 07 26.
Article in English | MEDLINE | ID: mdl-34105998

ABSTRACT

Multiple host proteins affect the gene expression of Kaposi's sarcoma-associated herpesvirus (KSHV) during latent and lytic replication. High-mobility group box 1 (HMGB1) serves as a highly conserved chromosomal protein inside the cell and a prototypical damage-associated molecular pattern molecule outside the cell. HMGB1 has been shown to play a pathogenic role in viral infectious diseases and to regulate the lytic replication of KSHV. However, its functional effects on the KSHV life cycle in KSHV-infected cells have not been fully elucidated. Here, we explored the role of intracellular and extracellular HMGB1 in KSHV virion production by employing CRISPR/Cas9-mediated HMGB1 knockout in the KSHV-producing iSLK BAC16 cell line. Intracellular HMGB1 formed complexes with various proteins, and the abundance of HMGB1-interacting proteins changed during latent and lytic replication. Moreover, extracellular HMGB1 was found to enhance lytic replication by phosphorylating JNK. Of note, the expression of viral genes was attenuated during lytic replication in HMGB1 knockout iSLK BAC16 cells, with significantly decreased production of infectious virions compared to that of wild-type cells. Collectively, our results demonstrate that HMGB1 is an important cellular cofactor that affects the generation of infectious KSHV progeny during lytic replication. IMPORTANCE The high-mobility group box 1 (HMGB1) protein has many intra- and extracellular biological functions with an intricate role in various diseases. In certain viral infections, HMGB1 affects the viral life cycle and pathogenesis. In this study, we explored the effects of HMGB1 knockout on the production of Kaposi's sarcoma-associated herpesvirus (KSHV). HMGB1 knockout decreased virion production in KSHV-producing cells by decreasing the expression of viral genes. The processes by which HMGB1 affects KSHV production may occur inside or outside infected cells. For instance, several cellular and viral proteins interacted with intracellular HMGB1 in a nucleosomal complex, whereas extracellular HMGB1 induced JNK phosphorylation, thereby enhancing lytic replication. Our results suggest that both intracellular and extracellular HMGB1 are necessary for efficient KSHV replication. Thus, HMGB1 may represent an effective therapeutic target for the regulation of KSHV production.


Subject(s)
Gene Expression Regulation, Viral , HMGB1 Protein/metabolism , Herpesvirus 8, Human/physiology , Virion/metabolism , Cell Line, Tumor , Gene Knockout Techniques , HMGB1 Protein/genetics , Herpesvirus 8, Human/genetics , Herpesvirus 8, Human/metabolism , Humans , Nucleosomes/metabolism , Promoter Regions, Genetic , Signal Transduction , Viral Proteins/genetics , Virus Activation , Virus Replication
10.
Vet Microbiol ; 256: 109048, 2021 May.
Article in English | MEDLINE | ID: mdl-33845333

ABSTRACT

The objective of this study was to assess protective efficacy of vaccination using CPD-attenuated chimeric PRRSV and Toll like receptor (TLR) agonists (HSP70 c-terminal domain and HSPX) as adjuvants through different inoculation routes. In this study, a chimeric PRRSV composed of two field isolates was synthesized and attenuated by CPD in NSP1 as described in the previous study. The infection of the CPD-attenuated chimeric PRRSV to pigs of 3 weeks-old showed no clinical signs without pathological lesions in necropsy, while it induced improved cross immunity between its parent strains. The TLR agonists were expressed in E. coli and purified to be used. In challenge experiment, pigs of 3 weeks-old were vaccinated using the CPD-attenuated chimeric virus with the prepared TLR agonists through intramuscular or intradermal route, following heterologous challenge after 4 weeks of vaccination. In results, intramuscular or intradermal inoculation of the CPD-attenuated chimeric virus demonstrated excellent protective efficacy against heterologous challenges. Importantly, intradermal inoculation with the TLR agonists enhanced protective effects as shown in the significantly increased level of PRRSV-specific IFN-γ-SCs and cytokines in sera, and the significant reduction of pathological lesion and viral load in lung. This study suggested that the intradermal inoculation of CPD-attenuated chimeric PRRSV plus TLR agonists should be more effective for protection of pigs against diverse PRRS field viruses.


Subject(s)
Porcine Reproductive and Respiratory Syndrome/prevention & control , Porcine respiratory and reproductive syndrome virus/immunology , Vaccination/veterinary , Viral Vaccines/administration & dosage , Adjuvants, Immunologic , Animals , Animals, Newborn , Chimera , Cytokines , Escherichia coli/genetics , Escherichia coli/metabolism , Injections, Intradermal/veterinary , Porcine Reproductive and Respiratory Syndrome/virology , Porcine respiratory and reproductive syndrome virus/genetics , Swine , Toll-Like Receptors/drug effects , Toll-Like Receptors/genetics , Toll-Like Receptors/metabolism , Vaccines, Attenuated/administration & dosage
11.
J Microbiol ; 59(5): 522-529, 2021 May.
Article in English | MEDLINE | ID: mdl-33877577

ABSTRACT

Extracellular vesicles (EVs) play a crucial role in cell-to-cell communication. EVs and viruses share several properties related to their structure and the biogenesis machinery in cells. EVs from virus-infected cells play a key role in virus spread and suppression using various loading molecules, such as viral proteins, host proteins, and microRNAs. However, it remains unclear how and why viruses regulate EV production inside host cells. The purpose of this study is to investigate the molecular mechanisms underlying EV production and their roles in Kaposi's sarcoma-associated herpesvirus (KSHV)-infected cells. Here, we found that KSHV induced EV production in human endothelial cells via Rab-27b upregulation. The suppression of Rab27b expression in KSHV-infected cells enhanced cell death by increasing autophagic flux and autolysosome formation. Our results indicate that Rab27b regulates EV biogenesis to promote cell survival and persistent viral infection during KSHV infection, thereby providing novel insights into the crucial role of Rab-27b in the KSHV life cycle.


Subject(s)
Extracellular Vesicles/metabolism , Herpesviridae Infections/metabolism , Herpesvirus 8, Human , rab GTP-Binding Proteins/metabolism , Autophagy , Cell Death , Cell Survival , Endothelial Cells/virology , Herpesviridae Infections/virology , Herpesvirus 8, Human/genetics , Humans , MicroRNAs/metabolism , Nanoparticles , Up-Regulation , Viral Proteins/metabolism
12.
BMC Vet Res ; 16(1): 452, 2020 Nov 23.
Article in English | MEDLINE | ID: mdl-33228643

ABSTRACT

BACKGROUND: Mycoplasma hyopneumoniae (M. hyopneumoniae), a representative pathogen causing swine enzootic pneumonia, generally infects piglets vertically. However, it is difficult to ascertain the M. hyopneumoniae infection state of sows due to limited detection methods. This report investigated sow herd stability by applying nested PCR to laryngeal swabs of suckling pigs, which is reportedly the most sensitive method. RESULTS: M. hyopneumoniae was detected in 14 farms (63.6%) and 127 piglets (6.5%). The prevalence of sows likely to transmit M. hyopneumoniae in herds (11.1%) was calculated. In addition, there was a significant difference in detection rates among farms depending on herd size, gilt replacement rate, acclimation method, and antibiotic usage, suggesting various parameters that influence sow stability. CONCLUSIONS: The results demonstrated that laryngeal swabs from suckling pigs have provided useful information regarding vertical transmission from sows in South Korean farm conditions. This result demonstrated that farms with larger herd sizes, higher gilt replacement rates, and a practice of naturally exposing gilts for acclimation had higher detection rates in weaning piglets, indicating an unstable sow infection state.


Subject(s)
Mycoplasma hyopneumoniae/isolation & purification , Pneumonia of Swine, Mycoplasmal/epidemiology , Pneumonia of Swine, Mycoplasmal/transmission , Animal Husbandry/methods , Animals , Female , Infectious Disease Transmission, Vertical , Pneumonia of Swine, Mycoplasmal/microbiology , Polymerase Chain Reaction/methods , Polymerase Chain Reaction/veterinary , Prevalence , Republic of Korea , Sus scrofa , Swine
13.
ACS Appl Mater Interfaces ; 12(38): 43090-43097, 2020 Sep 23.
Article in English | MEDLINE | ID: mdl-32862637

ABSTRACT

Selective emitters comprising plasmonic resonators have been exploited for cooling devices or infrared stealth technology. While selective emitters have been designed using odd-order resonances, even-order resonances also emit anisotropic thermal radiation signals. Thermal radiation by even-order resonances in selective emitters can be experimentally detected by thermal imaging cameras, and such thermal emissions often degrade the observability of infrared detectors, rendering them inapplicable to infrared stealth technology. Here, a selective emitter with extremely low thermal radiation signature in a dual-band range, a detection range by an infrared detector, is proposed with engineering anisotropic thermal radiation by even-order resonances. To minimize infrared signature in a dual-band range, the characterization of even-order resonances of gap plasmon metasurfaces is achieved based on vectorial diffraction within a relative error of 10%. Thermal radiation by even-order resonance has been shown to be highly directional and can be experimentally measured using mid-wave infrared images. Based on model prediction, the proposed selective emitter reduces mid-wave infrared signatures and long-wave infrared signatures by factors of 37.95 and 38.06, respectively, compared with those of blackbody surfaces. In addition, numerically confirmed thermal signature reduction and captured mid-wave infrared images indicate excellent thermal camouflage performance of the selective emitter with background medium. Thus, the characterization of even-order resonances provides a basis for the design of metasurfaces that can be employed for multispectral applications, especially infrared stealth technology.

14.
Virology ; 540: 172-183, 2020 01 15.
Article in English | MEDLINE | ID: mdl-31928999

ABSTRACT

Two type 2 field porcine reproductive and respiratory syndrome viruses (PRRSV) isolated from PRRS-affected swine farms were attenuated by de-optimization of codon pair bias in NSP1. In 3-week-old pigs infection, the attenuated viruses showed significantly lower replication ability than the original viruses without distinct clinical sign and pathological lesions, which were observed in pig infected with the original viruses. Regarding induction of PRRSV specific immunity, the level of the neutralizing antibodies as well as secretion of IFN-γ-SCs in PBMCs was not different between the attenuated viruses and the original viruses. More importantly, pigs infected with the attenuated viruses exhibited significant reduction in respiratory scores, viremia, macroscopic and microscopic lung lesion scores, and PRRSV-antigen with interstitial pneumonia against a heterologous challenge with a type 2 virulent strain. Conclusively, the viruses attenuated by CPD in this study demonstrated potential usefulness as vaccine strains to provide protective immunity against diverse virulent PRRSVs.


Subject(s)
Codon , Disease Resistance/genetics , Host-Pathogen Interactions/genetics , Porcine Reproductive and Respiratory Syndrome/virology , Porcine respiratory and reproductive syndrome virus/physiology , Viral Nonstructural Proteins/genetics , Animals , Antibodies, Neutralizing/immunology , Antibodies, Viral/immunology , Antigens, Viral/genetics , Antigens, Viral/immunology , Computational Biology/methods , Genome, Viral , Genomic Instability , Host-Pathogen Interactions/immunology , Interferon-gamma , Neutralization Tests , Phylogeny , Porcine Reproductive and Respiratory Syndrome/immunology , Porcine Reproductive and Respiratory Syndrome/pathology , Porcine respiratory and reproductive syndrome virus/classification , Swine , Viral Nonstructural Proteins/immunology , Viral Vaccines/administration & dosage , Viral Vaccines/immunology , Virulence , Virus Replication
15.
BMC Vet Res ; 15(1): 342, 2019 Oct 16.
Article in English | MEDLINE | ID: mdl-31619295

ABSTRACT

BACKGROUND: The objective of this study was to assess the efficacy of a trivalent vaccine mixture and compare it to the respective monovalent vaccines against Mycoplasma hyopneumoniae, porcine circovirus type 2 (PCV2), and porcine reproductive and respiratory syndrome virus (PRRSV). RESULTS: Pigs that were triple challenged with M. hyopneumoniae, PCV2, and PRRSV following vaccination with the trivalent vaccine mixture exhibited a significantly better growth performance when compared to unvaccinated and challenged pigs. A statistical difference was not found when comparing pig populations which were vaccinated with the trivalent vaccine followed by a triple challenge and pigs vaccinated with monovalent M hyopneumoniae vaccine followed by mycoplasmal single challenge in the following areas: M. hyopneumoniae nasal shedding, the number of M. hyopneumoniae-specific interferon-γ secreting cells (IFN-γ-SC), and mycoplasmal lung lesion scores. Pigs vaccinated with the trivalent vaccine mixture followed by a triple challenge resulted in a similar reduction of PCV2 viremia, an increase in the number of PCV2-specific IFN-γ-SC and reduction in interstitial lung lesion scores when compared to pigs vaccinated with a PCV-2 vaccine and challenged with PCV2 only. Lastly, there was a significant difference in the reduction of PRRSV viremia, an increase in PRRSV-specific IFN-γ-SC and a reduction of interstitial lung lesion scores between pigs vaccinated with the trivalent vaccine mixture followed by a triple challenge and pigs vaccinated with a monovalent PRRSV vaccine followed by PRRSV challenge only. CONCLUSION: The trivalent vaccine mixture was efficacious against a triple challenge of M. hyopneumoniae, PCV2, and PRRSV. The trivalent vaccine mixture, however, did not result in equal protection when compared against each respective monovalent vaccine, with the largest vaccine occurring within PRRSV.


Subject(s)
Circovirus/immunology , Mycoplasma hyopneumoniae/immunology , Porcine respiratory and reproductive syndrome virus/immunology , Swine Diseases/prevention & control , Vaccination/veterinary , Animals , Bacterial Vaccines/immunology , Circoviridae Infections/immunology , Circoviridae Infections/prevention & control , Female , Male , Pneumonia of Swine, Mycoplasmal/immunology , Pneumonia of Swine, Mycoplasmal/prevention & control , Porcine Reproductive and Respiratory Syndrome/immunology , Porcine Reproductive and Respiratory Syndrome/prevention & control , Sus scrofa , Swine , Swine Diseases/immunology , Swine Diseases/microbiology , Swine Diseases/virology , Vaccines, Combined/administration & dosage , Vaccines, Combined/immunology , Viral Vaccines/immunology
16.
Sci Rep ; 9(1): 8207, 2019 Jun 03.
Article in English | MEDLINE | ID: mdl-31160661

ABSTRACT

High-brightness light sources with nanoscale volume are required in nonlinear physics studies or various nanoscale engineering areas. Although several plasmonic devices, such as plasmonic nanofocusing, have been proposed for light concentration, the efficient enhancement of the nanofocusing device to get a bright light source is still limited owing to the inevitable Ohmic loss resulting from high field confinement on metallic surface. We propose the concept of dielectric nanofocusing by reversing the concept of conventional plasmonic nanofocusing and using a three-dimensional bowtie nanoaperture (3D BNA). The optical simulations demonstrate that the 3D BNA can achieve an intensity enhancement factor of 9.01 × 104. We calculate the dispersion relation for a tapered silver-SiNx-air waveguide to prove the possibility of focusing even for a high tapered angle. The theoretically calculated modal length can explain the origin of the high intensity enhancement by proving an energy flow from the dielectric layer to the air regime in dielectric nanofocusing. The performed optical and thermal simulations demonstrate that the 3D BNA can achieve a peak intensity of 6.21 PW/cm2 by avoiding the energy confinement around the metal. Our approach provides a new method for obtaining a high brightness light source.

17.
Vet Microbiol ; 223: 119-125, 2018 Sep.
Article in English | MEDLINE | ID: mdl-30173737

ABSTRACT

We have evaluated the cross-protection of a modified-live virus (MLV) vaccine based on porcine reproductive and respiratory syndrome virus (PRRSV)-2, against a heterologous PRRSV-1 challenge in late term pregnancy gilts. Gilts were vaccinated 42 days prior to breeding and then challenged intranasally with PRRSV-1 at 93 days of gestation. No local or systemic adverse effects related to vaccination were observed in the vaccinated gilts throughout the study. Vaccination resulted in a longer gestation period, a higher number of live-born and weaned piglets, and a significant decrease in the number of stillborn piglets compared to the unvaccinated group. The PRRSV-2 MLV vaccine was also able to significantly reduce PRRSV-1 viremia. At the time of PRRSV-1 challenge, vaccinated gilts had significantly higher PRRSV-1 specific interferon-γ secreting cells but low neutralizing antibody titers against PRRSV-1 compared to unvaccinated gilts. This correlated with a reduction of PRRSV-1 viremia, indicating that cell-mediated rather than humoral immunity played a role in PRRSV-1 clearance from the blood. Fetal thymic tissues from vaccinated pregnant gilts had fewer PRRSV-1 positive cells compared to unvaccinated gilts. Taken together these results indicate that vaccination of gilts with PRRSV-2 MLV vaccine can provide cross-protection against PRRSV-1 challenge and improve reproductive performance.


Subject(s)
Administration, Intranasal/veterinary , Porcine Reproductive and Respiratory Syndrome/prevention & control , Porcine respiratory and reproductive syndrome virus/immunology , Reproduction , Viral Vaccines/immunology , Animals , Antibodies, Neutralizing/immunology , Cross Protection , Female , Fetus/virology , Interferon-gamma/metabolism , Porcine Reproductive and Respiratory Syndrome/virology , Porcine respiratory and reproductive syndrome virus/genetics , Pregnancy , Stillbirth/veterinary , Swine , Vaccination/veterinary , Vaccines, Attenuated/immunology , Viremia/veterinary , Weaning
18.
Transbound Emerg Dis ; 65(6): 1641-1647, 2018 Dec.
Article in English | MEDLINE | ID: mdl-29877065

ABSTRACT

The objective of this study was to compare the severity of reproductive failure caused by either a single or a dual infection with porcine reproductive and respiratory syndrome virus (PRRSV)-1 and PRRSV-2 in late-term pregnancy gilts. Pregnant gilts were intranasally administered PRRSV-1, PRRSV-2 or both at 3 weeks before the expected farrowing date (93 days of gestation). Regardless of single and dual infection, PRRSV-infected pregnant gilts experienced premature farrowing (103-109 days of gestation) compared with negative control gilts which carried their pregnancy to full term (114-115 days of gestation). Pregnant gilts infected with only PRRSV-1 had a significantly (p < 0.05) higher number of genomic copies of PRRSV-1 in their blood compared with dually infected gilts. Additionally, stillborn foetuses and live-born piglets from pregnant gilts infected with only PRRSV-1 had a significantly (p < 0.05) higher number of PRRSV-1-positive cells per unit area of tissue sections examined, compared to pregnant gilts dually infected with PRRSV-1 and PRRSV-2. In contrast, pregnant gilts infected with only PRRSV-2 showed no difference in the number of genomic copies of PRRSV-2 compared with dually infected pregnant gilts and there were no significant differences in PRRSV-2-positive cells per unit area in tissues of stillborn foetuses and live-born piglets from pregnant gilts infected with PRRSV-2 only compared with dually infected gilts. Interestingly, even though PRRSV-2 was shown to replicate more efficiently compared with PRRSV-1 in dually infected pregnant gilts, neither PRRSV type was able to exacerbate reproductive failure in pregnant gilts already dually infected with PRRSV-1 and PRRSV-2. Our results suggest that the severity of reproductive failure is similar between dual (PRRSV-1 and PRRSV-2) and single infection (PRRSV-1 or PRRSV-2).


Subject(s)
Porcine Reproductive and Respiratory Syndrome/pathology , Porcine respiratory and reproductive syndrome virus/pathogenicity , Pregnancy Complications, Infectious/veterinary , Pregnancy, Animal , Administration, Intranasal , Animals , Female , Fetal Death , Fetus/virology , Porcine Reproductive and Respiratory Syndrome/virology , Porcine respiratory and reproductive syndrome virus/genetics , Porcine respiratory and reproductive syndrome virus/isolation & purification , Pregnancy , Pregnancy Complications, Infectious/virology , Stillbirth , Sus scrofa , Swine
19.
Can J Vet Res ; 82(2): 146-153, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29755195

ABSTRACT

A novel porcine circovirus type 2 (PCV2) peptide vaccine comprised of a consensus capsid (Cap) protein domain encoded by open reading frame 2 was developed to control PCV2 infection. The efficacy of the vaccine was evaluated against a commercial baculovirus-expressed recombinant PCV2 subunit vaccine based on the Cap protein. The amino acid sequence of this Cap protein was designed based on the alignment of amino acid sequences from different isolates from Europe, North America, and Asia. The vaccine was evaluated in either phosphate-buffered saline or adjuvanted with aluminum hydroxide, cobalt oxide, or liposome. Overall the PCV2 peptide vaccine was less efficacious against PCV2 challenge compared with the commercial PCV2 vaccine. The peptide vaccine was the most efficacious when liposome was used as an adjuvant, significantly (P < 0.05) reducing viremia while increasing the levels of neutralizing antibodies and interferon-γ secreting cells. This suggests, in the presence of liposome, the peptide vaccine was able to elicit both humoral and cellular immune responses.


Un nouveau vaccin peptidique contre le circovirus porcin de type 2 (CVP2) contenant un domaine d'une protéine consensus de la capside (Cap) codé par le cadre de lecture ouvert 2 fut développé pour limiter l'infection par CVP2. L'efficacité du vaccin fut évaluée versus celle d'un vaccin commercial CVP2 sous-unitaire recombinant s'exprimant dans un baculovirus et basé sur la protéine Cap. La séquence en acides aminés de cette protéine Cap a été élaborée basée sur l'alignement des séquences d'acides aminés provenant de différents isolats d'Europe, d'Amérique du Nord, et d'Asie. Le vaccin a été évalué dans soit de la saline tamponnée avec du phosphate ou dans un adjuvant contenant de l'hydroxyde d'aluminium, d'oxyde de cobalt, ou des liposomes. Globalement, le vaccin peptidique CVP2 était moins efficace contre une infection défi par CVP2 comparativement au vaccin commercial. Le vaccin peptidique était le plus efficace lorsque les liposomes étaient utilisés comme adjuvant, réduisant de manière significative (P < 0,05) la virémie tout en augmentant la quantité d'anticorps neutralisants et de cellules secrétant de l'interféron γ. Ceci suggère qu'en présence de liposomes le vaccin lipidique était en mesure d'induire des réponses immunitaires humorale et cellulaire.(Traduit par Docteur Serge Messier).


Subject(s)
Circoviridae Infections/veterinary , Circovirus/classification , Swine Diseases/prevention & control , Viral Vaccines/immunology , Animals , Antibodies, Neutralizing/blood , Circoviridae Infections/prevention & control , Circoviridae Infections/virology , Circovirus/immunology , DNA, Viral/blood , Interferon-gamma/metabolism , Swine , Swine Diseases/virology , Vaccines, Subunit/immunology , Vaccines, Synthetic/immunology
20.
Vet Rec ; 182(21): 602, 2018 05 26.
Article in English | MEDLINE | ID: mdl-29545353

ABSTRACT

The objective of this field study was to evaluate the reproductive performance of sows after vaccination with a porcine reproductive and respiratory syndrome (PRRS) subunit vaccine (PRRSFREE PRRS subunit vaccine, Reber Genetics, Taiwan, Republic of China) under field conditions. The study was performed in three farms with endemic infections with both PRRS virus (PRRSV)-1 and PRRSV-2, a situation representative of most Korean farms. Pregnant sows were immunised intramuscularly with 2.0 ml of the PRRS subunit vaccine at 58 and 79 days of gestation (eight and five weeks antepartum) according to the manufacturer's recommendation. Vaccination did not result in any observed adverse reaction. Vaccinated sows exhibited a significant improvement in reproductive performance (reduction of abortions) and litter characteristics (increase of weaned pigs) compared with unvaccinated sows. Vaccinated sows had significantly (P<0.05) higher PRRSV ELISA sample/positive ratio and number of PRRSV-specific interferon-γ-secreting cells compared with the unvaccinated control group. The results of this study demonstrate that the PRRS subunit vaccine can improve the reproductive performance of sows in farms with endemic PRRSV infection.


Subject(s)
Endemic Diseases/veterinary , Porcine Reproductive and Respiratory Syndrome/prevention & control , Reproduction/physiology , Viral Vaccines/administration & dosage , Animals , Endemic Diseases/prevention & control , Farms , Female , Porcine Reproductive and Respiratory Syndrome/epidemiology , Porcine Reproductive and Respiratory Syndrome/virology , Porcine respiratory and reproductive syndrome virus/genetics , Porcine respiratory and reproductive syndrome virus/isolation & purification , Pregnancy , RNA, Viral/blood , Swine , Taiwan/epidemiology , Vaccines, Subunit/administration & dosage
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