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2.
Vaccine ; 42(6): 1292-1299, 2024 Feb 27.
Article in English | MEDLINE | ID: mdl-38296705

ABSTRACT

BACKGROUND: The emergence and rapid spread of new mutant strains of SARS-CoV-2 necessitate the development of a new generation vaccine capable of neutralizing a broad range of variants. When the SARS-CoV-2 Omicron variant emerged, individuals in China had already received an inactivated (INA) or a type 5 adenovirus-vectored (Ad5) SARS-CoV-2 vaccine targeting the wild-type virus. We have recently developed a bivalent recombinant type 5 vaccine targeting both the wild-type strain and the Omicron variant (Ad5-nCoV/O). The objectives of this study were to assess the immunogenicity of the bivalent vaccine as a booster against both the wild type and the Omicron variant. METHODS: In the single immunization model, mice received one intramuscular immunization with monovalent or bivalent Ad5-vectored vaccines targeting both wild-type SARS-CoV-2 and Omicron variants. In the prime-boost model, mice were primed intramuscularly with an INA or Ad5-vectored vaccine targeting wild-type SARS-CoV-2, and then boosted intramuscularly or intranasally with heterologous or homologous INA or monovalent or bivalent Ad5-vectored vaccines targeting both wild-type SARS-CoV-2 and Omicron variants. The vaccine-induced antibody responses and cellular immune responses were measured using ELISA, pseudovirus-based neutralization assays, the intracellular cytokine staining (ICS) and ELISpot. RESULTS: Single-dose prime vaccination with the monovalent and bivalent vaccines elicited robust antibody responses and CD4 + and CD8 + cellular responses against the spike protein of WT and Omicron SARS-CoV-2. Both intramuscular and intranasal boost vaccination with the bivalent Ad5-nCoV/O following a prime with INA or Ad5-vectored vaccines induced strong serum neutralization antibody responses to both wild type and Omicron variants. A heterologous prime-boost vaccination elicited greater neutralization antibody responses than a homologous prime-boost vaccination when mice were boosted with Ad5-vectored vaccines following a prime with INA. Intranasal boost also resulted in significant mucosal IgA responses. CONCLUSION: The bivalent vaccine Ad5-nCoV/O exhibited robust immunogenicity, inducing broad-spectrum cross-neutralizing antibodies and cellular immune responses against both wild type and Omicron variants of SARS-CoV-2. The results demonstrated the potential of the bivalent vaccine in addressing the challenges posed by emerging SARS-CoV-2 Omicron variants.


Subject(s)
COVID-19 Vaccines , COVID-19 , Animals , Humans , Mice , Vaccines, Combined , Disease Models, Animal , SARS-CoV-2 , COVID-19/prevention & control , Vaccination , Enzyme-Linked Immunospot Assay , Adenoviridae/genetics , Antibodies, Neutralizing , Antibodies, Viral , Immunogenicity, Vaccine
3.
Emerg Microbes Infect ; 11(1): 1550-1553, 2022 Dec.
Article in English | MEDLINE | ID: mdl-35604772

ABSTRACT

In order to overcome the pandemic of COVID-19, messenger RNA (mRNA)-based vaccine has been extensively researched as a rapid and versatile strategy. Herein, we described the immunogenicity of mRNA-based vaccines for Beta and the most recent Omicron variants. The homologous mRNA-Beta and mRNA-Omicron and heterologous Ad5-nCoV plus mRNA vaccine exhibited high-level cross-reactive neutralization for Beta, original, Delta, and Omicron variants. It indicated that the COVID-19 mRNA vaccines have great potential in the clinical use against different SARS-CoV-2 variants.


Subject(s)
COVID-19 , SARS-CoV-2 , Antibodies, Neutralizing , Antibodies, Viral , BNT162 Vaccine , COVID-19/prevention & control , Humans , RNA, Messenger/genetics , SARS-CoV-2/genetics , Vaccines, Synthetic , mRNA Vaccines
4.
Commun Biol ; 4(1): 364, 2021 03 19.
Article in English | MEDLINE | ID: mdl-33742082

ABSTRACT

The fruit fly, Drosophila melanogaster, has been used as a model organism for the molecular and genetic dissection of sleeping behaviors. However, most previous studies were based on qualitative or semi-quantitative characterizations. Here we quantified sleep in flies. We set up an assay to continuously track the activity of flies using infrared camera, which monitored the movement of tens of flies simultaneously with high spatial and temporal resolution. We obtained accurate statistics regarding the rest and sleep patterns of single flies. Analysis of our data has revealed a general pattern of rest and sleep: the rest statistics obeyed a power law distribution and the sleep statistics obeyed an exponential distribution. Thus, a resting fly would start to move again with a probability that decreased with the time it has rested, whereas a sleeping fly would wake up with a probability independent of how long it had slept. Resting transits to sleeping at time scales of minutes. Our method allows quantitative investigations of resting and sleeping behaviors and our results provide insights for mechanisms of falling into and waking up from sleep.


Subject(s)
Activity Cycles , Behavior, Animal , Drosophila melanogaster/physiology , Sleep Stages , Age Factors , Animals , Locomotion , Models, Theoretical , Rest , Time Factors , Video Recording
5.
Sci Adv ; 5(5): eaaw5141, 2019 05.
Article in English | MEDLINE | ID: mdl-31131327

ABSTRACT

Mechanoreception detects physical forces in the senses of hearing, touch, and proprioception. Here, we show that labellar mechanoreception wires two motor circuits to facilitate and terminate Drosophila feeding. Using patch-clamp recordings, we identified mechanosensory neurons (MSNs) in taste pegs of the inner labella and taste bristles of the outer labella, both of which rely on the same mechanoreceptor, NOMPC (no mechanoreceptor potential C), to transduce mechanical deflection. Connecting with distinct brain motor circuits, bristle MSNs drive labellar spread to facilitate feeding and peg MSNs elicit proboscis retraction to terminate feeding. Bitter sense modulates these two mechanosensory circuits in opposing manners, preventing labellar spread by bristle MSNs and promoting proboscis retraction by peg MSNs. Together, these labeled-line circuits enable labellar peg and bristle MSNs to use the same mechanoreceptors to direct opposing feeding actions and differentially integrate gustatory information in shaping feeding decisions.


Subject(s)
Drosophila/physiology , Feeding Behavior , Mechanoreceptors/physiology , Motor Neurons/physiology , Animals , Animals, Genetically Modified , Behavior, Animal , Biotin/analogs & derivatives , Biotin/physiology , Brain/physiology , Drosophila Proteins/physiology , Green Fluorescent Proteins , Mechanotransduction, Cellular , Neurons/physiology , Optogenetics , Patch-Clamp Techniques , RNA Interference , Stress, Mechanical , Taste
6.
Pharm Biol ; 49(6): 608-13, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21554003

ABSTRACT

CONTEXT: Phosphonoformate sodium (PFS) has been used as an anti-herpesvirus drug; nevertheless, studies of the use of PFS for treatment of pseudorabies herpesvirus (PrV) infection in the veterinary setting have not been widely reported. OBJECTIVE: The present study aimed to analyze the inhibitory effect of PFS on cell infection and apoptosis induced by PrV. MATERIALS AND METHODS: The infectivity of PrV was determined by plaque assays when PFS was applied to the virus, to the virus-infected cells, and to the cells prior to infection. PCR amplifying DNA polymerase, gE, gG, and gD genes of PrV was performed. PrV-induced cell apoptosis was analyzed by immunofluorescence and flow cytometry. RESULTS: PFS inhibits cell infection by PrV. Addition of the drug decreased the number of apoptotic cells. Amplification of DNA polymerase and other viral structural genes detected in this study by PCR was reduced, because there were fewer viral DNA copies being made in the presence of the drug. The drug has an inhibitory effect on cell apoptosis induced by PrV. DISCUSSION AND CONCLUSION: PFS has inhibitory effects on cell infection by PrV, which may be used as an anti-PrV agent or combined with other anti-PrV agents. PrV-induced cell apoptotic cells and viral DNA copies decreased in the presence of the PFS.


Subject(s)
Antiviral Agents/pharmacology , Foscarnet/pharmacology , Herpesvirus 1, Suid/drug effects , Herpesvirus 1, Suid/growth & development , Viral Plaque Assay/methods , Animals , Apoptosis/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Chlorocebus aethiops , DNA, Viral/drug effects , Dose-Response Relationship, Drug , Herpesvirus 1, Suid/genetics , Microbial Sensitivity Tests/methods , Vero Cells
7.
Pharm Biol ; 49(2): 161-6, 2011 Feb.
Article in English | MEDLINE | ID: mdl-20942608

ABSTRACT

CONTEXT: Pseudorabies herpesvirus (PrV) belongs to the Alphaherpesvirinae. Piglets infected with PrV die within a few days. Development of effective antiviral agents is one alternative or complementary method to prevent PrV infection. Houttuynia cordata Thunb. (Saururaceae), H. cordata, a traditional Chinese medicine, is often used to relieve lung abnormal symptoms, infectious disease, refractory hemoptysis and malignant pleural effusion in China. OBJECTIVE: The present study aimed to investigate the effect of H. cordata injection on cell infection by PrV using Vero cells (a monkey kidney cell line) and swine testis cells (ST) as models. MATERIALS AND METHODS: The infectivity of PrV was determined by plaque assays when H. cordata was applied to the virus, to the virus infected cells, and to the cells prior to infection. The genomic DNA copies post-drug treatment were confirmed by PCR and reverse transcription PCR. The cell apoptosis caused by the virus was analyzed. RESULTS: H. cordata efficiently inhibited cell infection after incubating the drug with PrV. Nevertheless, H. cordata was more efficient in Vero cells than in ST cells in terms of its inhibitory effect. Low-dosage drug inhibited cell apoptosis induced by PrV; nevertheless, high-dosage drug alone resulted in cell apoptosis. DISCUSSION AND CONCLUSION: H. cordata has a direct inhibitory activity against PrV in vitro. H. cordata may be used as an anti-PrV agent or combined with other anti-PrV agents. PrV infection induces cell apoptosis and H. cordata inhibits cell infection. The optimal administration dosage of H. cordata should be taken into account in the future, because high-dosage H. cordata alone causes cell apoptosis.


Subject(s)
Antiviral Agents/pharmacology , Apoptosis/drug effects , Drugs, Chinese Herbal/pharmacology , Pseudorabies/drug therapy , Animals , Antiviral Agents/administration & dosage , Chlorocebus aethiops , Dose-Response Relationship, Drug , Drugs, Chinese Herbal/administration & dosage , Houttuynia , Male , Polymerase Chain Reaction/methods , Reverse Transcriptase Polymerase Chain Reaction/methods , Swine , Testis/cytology , Testis/drug effects , Testis/virology , Vero Cells
8.
Antiviral Res ; 85(2): 346-53, 2010 Feb.
Article in English | MEDLINE | ID: mdl-19879899

ABSTRACT

Diammonium glycyrrhizin (DG), a salt from glycyrrhizinate (GL) that is a major active component of licorice root extract with various pharmacological activities was investigated for its inhibitory effect on pseudorabies virus (PrV) infection. In parallel, lithium chloride (LiCl), a chemical reagent with potential antiviral activity was compared with DG for their inhibitory ability against PrV infection in vitro. Virus plaque-reduction assays, PCR and RT-PCR analysis indicated that both drugs inhibited cell infection by PrV. Moreover, addition of the drugs resulted in fewer apoptotic cells during PrV infection.


Subject(s)
Antiviral Agents/pharmacology , Glycyrrhizic Acid/pharmacology , Herpesvirus 1, Suid/drug effects , Lithium Chloride/pharmacology , Animals , Apoptosis , Cell Survival , Chlorocebus aethiops , Glycyrrhiza/chemistry , Molecular Structure , Plant Extracts/pharmacology , Plant Roots/chemistry , Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Vero Cells , Viral Plaque Assay
9.
J Biotechnol ; 141(1-2): 91-6, 2009 Apr 20.
Article in English | MEDLINE | ID: mdl-19428736

ABSTRACT

Porcine aminopeptidase N (pAPN) is a cellular membrane protein and a functional receptor for porcine coronaviruses. Here, we describe the heterologous expression of pAPN without signal peptide in BL21(DE3)pLysS host cells. The Escherichia coli (E. coli) harboring the recombinant construct was efficiently induced to express the pAPN protein at a high level. The most optimal expression profile for pAPN expression was investigated. By inoculating a rabbit with the purified pAPN, a high tittered specific antibody was achieved. Biologically, the antibody reacted with either pAPN-expressing E. coli or native pAPN on the surface of swine testis cells. The pAPN and its specific antibody blocked transmissible gastroenteritis coronavirus infection in vitro. Furthermore, the localization of pAPN on the small intestine of swine was analyzed by immunohistochemistry.


Subject(s)
CD13 Antigens/biosynthesis , CD13 Antigens/metabolism , Escherichia coli/metabolism , Recombinant Proteins/biosynthesis , Recombinant Proteins/metabolism , Animals , CD13 Antigens/genetics , Cell Line , Coronavirus/physiology , Enzyme-Linked Immunosorbent Assay , Escherichia coli/genetics , Gene Expression , Immunohistochemistry , Male , Recombinant Proteins/genetics , Swine
10.
Avian Pathol ; 38(3): 215-21, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19468938

ABSTRACT

The effects of glycyrrhizin diammonium (GD) and lithium chloride (LiCl) on cell infection by avian infectious bronchitis virus (IBV) were investigated using cytopathic effect observation, plaque-reduction assay and reverse transcriptase-polymerase chain reaction. The anti-viral effect of GD and LiCl on virus, on virus-infected cells or on cells pre-treated by both drugs was analysed, respectively. Our results showed that GD had a direct antiviral activity, leading to complete inhibition of cell infection. The cell infection was not alleviated by either pre-treatment of cells with GD or addition of the drug post infection, confirming that the inhibitory effect of GD, unlike LiCl, on IBV is a viral factor, rather than a cellular factor. The inhibitory effect of both drugs was confirmed by infecting primary chicken embryo kidney cells. In addition, apoptosis of infected cells was positively related with cytopathic effect and could be inhibited by effective drug treatment. Our data indicate that GD and LiCl have potential to prevent IBV infection in vitro through different antiviral mechanisms. The data are helpful for using antivirals efficiently.


Subject(s)
Antiviral Agents/pharmacology , Glycyrrhizic Acid/pharmacology , Infectious bronchitis virus/drug effects , Infectious bronchitis virus/genetics , Lithium Chloride/pharmacology , Virus Internalization/drug effects , Animals , Apoptosis/drug effects , Chick Embryo , Chlorocebus aethiops , DNA Primers/genetics , Reverse Transcriptase Polymerase Chain Reaction , Vero Cells
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