1.Progress on mRNA vaccine for the prevention of major infectious diseases in humans and animals.
Jia-Qi GAO ; Xian-Jun ZHAO ; Jin-Lian HUA
Acta Physiologica Sinica 2023;75(5):647-658
A large number of studies have demonstrated that mRNA vaccine has been characterized as a technique with good safety, strong immunogenicity and high developmental potential, which makes it have broad prospects in immunotherapy. In recent years, the stability and in vivo delivery efficiency of mRNA vaccines have been largely addressed by the progresses in mRNA engineering and delivery innovation. And some mRNA vaccines are now clinical approved or in preclinical trials. Here, we summarize current knowledge on the research advances, technology, and application in major infectious diseases in humans and animals of mRNA vaccines, with the aim to provide a reference for improving the development of novel mRNA vaccines.
Animals
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Humans
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Communicable Diseases
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Vaccines, Synthetic/genetics*
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mRNA Vaccines
4.Advance in research on novel dengue vaccine.
Chinese Journal of Virology 2011;27(3):315-318
5.Applications of reverse genetics in studying classical swine fever virus.
Dafei LIU ; Yuan SUN ; Huaji QIU
Chinese Journal of Biotechnology 2009;25(10):1441-1448
Classical swine fever (CSF), caused by classical swine fever virus (CSFV), has been epidemic or endemic in many countries, and causes great economical losses to pig industry worldwide. Attenuated vaccines (such as C-strain) have played an important role in the control of CSF. Recently some new phenomena appear, such as atypical and persistent infections of CSF, immunization failure and so on. Meanwhile, eradication programs have been implemented in many countries, restricting the widespread applications of attenuated vaccines. Thus, currently the priority is to strengthen the research in pathogenesis and transmission mechanisms, as well as to develop marker vaccines. Recently, the applications of reverse genetics technology open up a new way for research of structure and function of CSFV proteins and development of novel vaccines against CSF. This review focuses on the progress of applications of reverse genetics in the functional analysis and marker vaccine development of CSFV, and also discusses the problems confronted now and prospective aspects in the study of CSFV.
Classical swine fever virus
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genetics
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Cloning, Molecular
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Genetics, Microbial
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methods
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RNA, Viral
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genetics
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Recombination, Genetic
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Vaccines, Synthetic
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biosynthesis
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immunology
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Viral Vaccines
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biosynthesis
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genetics
6.Expression of influenza A3 virus (H3N2) M2 gene in vaccinia virus Tiantan strain.
Wen-Ling WANG ; Bao-Ying HUANG ; Yao DENG ; Xiu-Ping WANG ; Wen-Jie TAN ; Li RUAN
Chinese Journal of Virology 2007;23(5):377-383
To construct a recombinant vaccinia virus RVJ1175M2 expressing influenza A3 virus (H3N2) M2 gene, full length gene encoding influenza virus (H3N2) M2 protein was amplified with PCR and cloned into plasmid pJSC1175 which was used for homologous recombination with vaccinia virus Tiantan strain. Along with this, a recombinant vaccinia virus RVJ1175M2 containing the M2 gene was subsequently constructed. It was identified by PCR that the gene of M2 protein was inserted into the TK locus of vaccinia virus Tiantan strain correctly and M2 protein was expressed by recombinant vaccinia virus RVJ1175M2 effectively. Two electrophoretic bands of M2 protein expressed by the infected HeLa cells, one of 15kD and the other of 13kD in accordance with related documents, was deteced by Western-blot. M2 protein distributing on the surface of the infected cells was demonstrated by immunofluorescence and flow cytometry. The results suggested that recombinant vaccinia virus RVJ1175M2 could express M2 protein effectively, this laid a foundation for comparative research on the immune effect of universal vaccine of influenza virus with different kinds of vaccine expressing M2 protein.
HeLa Cells
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Humans
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Influenza A Virus, H3N2 Subtype
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genetics
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Influenza Vaccines
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immunology
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Polymerase Chain Reaction
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Recombinant Proteins
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biosynthesis
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Vaccines, Synthetic
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immunology
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Vaccinia virus
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genetics
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Viral Matrix Proteins
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genetics
7.Construction and characterization of a pseudorabies virus TK-/gG- mutant.
Xiao-Juan XU ; Gao-Yuan XU ; Huan-Chun CHEN ; Zheng-Fei LIU ; Qi-Gai HE
Chinese Journal of Biotechnology 2004;20(4):532-535
To construct a TK-/gG- mutant of pseudorabies virus, the gG-detected transfer vector pUSKKBB and genomic DNA of pseudorabies virus TK-/gG-/LacZ+ were co-transfected into IBRS-2 cells. Transfection progeny were plated onto PK-15 cells and incubated for 2 days under methylcellulose. Then the overlay was removed and replaced by 1% low melting point agarose in DMEM supplemented with 150 microg/mL X-gal. After 2 days, white plaques were screened for and purified 4 times. By PCR amplification of gG-deleted gene and LacZ gene, a recombinant virus with TK-/gG- phenotype was confirmed. Sequence of the PCR product revealed that there were 1,176 bp detection in gG gene of the PRV TK-/gG- mutant. Amplifying the gG-deleted gene of different generations of the TK-/gG- mutant showed that the mutant was stable within PK-15 cells. TCID50 assay indicated that the recombinant virus grows well on PK-15 cells. The mice immunized with the TK-/gG- virus showed no sign of abnormality. As a control, all mice inoculated with PRV strain died from the infection. All mice that received TK-/gG- survived after a lethal PRV challenge. However none of the mice injected with phosphate-buffer saline (PBS) survived from the challenge. The above results demonstrated that the recombinant virus could be a candidate marker vaccine strain for eradicating pseudorabies in pig herds.
Animals
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Herpesvirus 1, Suid
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genetics
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pathogenicity
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Mice
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Mice, Inbred BALB C
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Mutation
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Pseudorabies Vaccines
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immunology
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Swine
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Thymidine Kinase
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genetics
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Vaccines, Synthetic
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Viral Envelope Proteins
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genetics
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immunology
8.Construction, screening and immunogenicity of the recombinant poxvirus vaccine rVTTδTK-RBD against SARS-CoV-2.
Renshuang ZHAO ; Yilong ZHU ; Chao SHANG ; Jicheng HAN ; Zirui LIU ; Zhiru XIU ; Shanzhi LI ; Yaru LI ; Xia YANG ; Xiao LI ; Ningyi JIN ; Xin JIN ; Yiquan LI
Chinese Journal of Cellular and Molecular Immunology 2024;40(1):19-25
Objective To construct a recombinant poxvirus vector vaccine, rVTTδTK-RBD, and to evaluate its safety and immunogenicity. Methods The receptor-binding domain (RBD) gene was synthesized with reference to the gene sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and was inserted into the polyclonal site of the self-constructed recombinant plasmid pSTKE, to construct the recombinant poxvirus shuttle vector pSTKE-RBD. This was then transfected into BHK-21 cells pre-infected with the vaccinia virus Tiantan strain (VTT). The recombinant poxvirus rVTTδTK-RBD was successfully obtained after several rounds of fluorescence phage screening. The effect of rVTTδTK-RBD on the body mass of BALB/c mice was detected after immunizing mice by intra-nasal vaccination. The levels of specific and neutralizing antibodies produced by rVTTδTK-RBD on BALB/c mice were analyzed after immunizing mice intramuscularly. The effect of rVTTδTK-RBD on T cell subsets in BALB/c mice was detected by flow cytometry. Results Through homologous recombination, enhanced green fluorescent protein (EGFP) screening marker, and multiple rounds of fluorescent phosphorescence phage screening, a recombinant poxvirus rVTTδTK-RBD, expressing RBD with deletions in the thymidine kinase (TK) gene, was successfully obtained, which was validated by PCR. The in vivo experiments on BALB/c mice showed that rVTTδTK-RBD was highly immunogenic against SARS-CoV-2 and significantly reduced toxicity to the body compared to the parental strain VTT. Conclusion The recombinant poxvirus vaccine rVTTδTK-RBD against SARS-CoV-2 is successfully constructed and obtained, with its safety and immunogenicity confirmed through various experiments.
Animals
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Mice
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SARS-CoV-2/genetics*
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COVID-19
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Vaccines, Synthetic/genetics*
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Genes, Reporter
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Bacteriophages
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Mice, Inbred BALB C
9.The immunogenicity in mice enhanced significantly via prime-boost vaccination with DNA-based or recombinant vaccinia(Tiantan) viral-based H5N1 vaccine candidates containing multi-structural antigens.
Wen WANG ; Hong CHEN ; Yao DENG ; Yang YANG ; Xiao YIN ; Min WANG ; Jian-Fang ZHOU ; Yue-Ling SHU ; Li RUAN ; Wen-Jie TAN
Chinese Journal of Virology 2011;27(6):594-598
This study aimed to develop an effective experimental vaccine against highly pathogenic H5N1 Avian Influenza (HPAI) virus and to optimize their immunization programs. As reported previously, various DNA-based or recombinant vaccinia viral(Tiantan)-based H5N1 vaccine candidates, which containing a single cistronic construct (HAop, or NAop) or a bicistronic construct (HAop/M2 or NAop/M1) of H5N1 influenza virus (Anhui strain) were constructed and characterized in our lab. In this study, we further analysed the immunogenicity in mice of these vaccine candidates by various protocols (single or combined immunization). Our results showed that: comparing with immunization with DNA-based or rTTV-based H5N1 vaccine only, combined DNA-based with rTTV-based H5N1 vaccine immunization via prime-boost strategy enhanced immune response significantly against multi-H5N1 antigens detected by hemagglutination inhibition (HI) assay, NA- or M1- or M2-specific antibody detection, and micro-neutralizing antibody test and IFN-gamma ELISpot assay. Priming with DNA-based vaccine induced higher level of humoral response against HA or NA antigen than priming with rTTV-based vaccine; In contract, M1 and M2-specific antibody levels were higher among that of priming with rTTV -based vaccine. These findings provide a basis for further development of novel H5N1 vaccines and for the optimization of the immunization programs of combined multi-antigens vaccine candidates.
Animals
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Antigens, Viral
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genetics
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immunology
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Female
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Immunization
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methods
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Influenza A Virus, H5N1 Subtype
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genetics
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immunology
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Influenza Vaccines
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genetics
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immunology
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Mice
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Mice, Inbred BALB C
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Vaccination
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methods
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Vaccines, DNA
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genetics
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immunology
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Vaccines, Synthetic
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genetics
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immunology
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Vaccinia
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genetics
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immunology
10.Characterization of attenuated Salmonella C500 strain with a delta asd mutant and use as an Asd+ balanced-lethal host-vector system.
Zhanqin ZHAO ; Yindi XU ; Bin WU ; Xiangchao CHENG ; Yinju LI ; Xibiao TANG ; Chunjie ZHANG ; Huanchun CHEN
Chinese Journal of Biotechnology 2009;25(1):29-36
Salmonella enterica serovar Choleraesuis strain C500 is a live, attenuated vaccine that has been used in China for over 40 years to prevent piglet paratyphoid. The objective of this study was to evaluate the potential of attenuated Salmonella enterica serovar Choleraesuis C500 strain with a delta asd mutant as an effective live vaccine vector by the Asd+ balanced-lethal host-vector system. Here, we compared the characteristics of S. enterica serovar Choleraesuis delta asdC500 strain with the parent C500 strain, including phenotype, growth rate, virulence, safety, and expression for heterologous antigen. The mean generation times of delta asdC500 mutant, the vector control delta asdC500 (pYA3493), and the parent avirulent C500 vaccine strain in Luria broth were 30.7, 28.1, and 27.9 min, respectively. The fermentation patterns of theses three strains on different carbohydrates, and the levels of production of H2S, were similar. The O and H antigens of delta asdC500 mutant, delta asdC500 (pYA3493) and delta asdC500 (pYA-F1P2) were 6,7:C:1,5, identical to the parent strain C500. By the method of Reed and Muench, groups of mice were challenged by the intraperitoneal route with different amounts of delta asdC500 (pYA3493) or the parent C500 strain, and the virulence of delta asdC500 (pYA3493) with LD50 of 1.1 x 10(7) CFU was a little lower than C500 with LD50 of 4.4 x 10(6) CFU. All piglets inoculated with delta asdC500 (pYA3493) or C500 survived, and no signs of disease were observed during the entire experimental period. No major differences were found in these two groups. In addition, the recombinant pYA-F1P2 plasmid was very stable in the recombinant delta asdC500 (pYA-F1P2) strain, which expressed secretorily a large amount of the recombinant filamentous hemagglutinin type I domain and pertactin region 2 domain antigen (rF1P2) of Bordetella bronchiseptica. In this study, we have shown that the delta asdC500 mutant had a series of biological characteristics similar to the parent vaccine strain C500. Furthermore, the strain could express secretorily a large amount of heterologous antigen. It is likely that this Salmonella expression and delivery system could be easily adapted to develop multivalent recombinant Salmonella vaccines against infectious agents using the Asd+ balanced-lethal host-vector system.
Amino Acid Oxidoreductases
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genetics
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Animals
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Bacterial Proteins
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genetics
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Gene Deletion
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Genetic Vectors
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Mice
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Mutation
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Salmonella Vaccines
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genetics
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immunology
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Salmonella enterica
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genetics
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immunology
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pathogenicity
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Swine
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Transduction, Genetic
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Vaccines, Attenuated
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genetics
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immunology
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Vaccines, Synthetic
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genetics
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immunology
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Virulence