2.Strategies for screening protective viral antigens and their applications in the development of novel vaccines.
Dailang ZHONG ; Tao WANG ; Rui LUO ; Hua-Ji QIU ; Yuan SUN
Chinese Journal of Biotechnology 2022;38(8):2857-2871
With the development of vaccine research and development technologies, novel vaccines have been widely used in the prevention of various infectious diseases. Due to the excellent safety, novel vaccines have unique advantages in the application of vaccines against virulent pathogens. The major premise of developing novel vaccines is to screen protective antigens. With the development of various omics research, cutting-edge bioinformatics tools for eukaryotes have been well developed, while the much simpler structure of viruses compared with eukaryotic cells corresponds to relatively simple research methods. Strategies for screening protective antigens need to combine the advantages of both bioinformatics methods and traditional molecular biology methods. In this review, the strategies for screening virus protective antigens were discussed from the perspective of host and virus, and a series of bioinformatics tools developed based on eukaryotic cells that may be used for screening protective antigens were listed. This review also summarized the cases of using protective antigens to design novel vaccines, in order to better understand the strategies for screening virus protective antigens and facilitate the research and development of novel vaccines.
Antigens
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Antigens, Viral/genetics*
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Computational Biology
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Research
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Vaccines
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Viral Vaccines/genetics*
3.Post-zoster Granuloma.
Hyang Joon PARK ; Jeong Heon LEE ; You Chan KIM ; Yong Woo CINN
Annals of Dermatology 1998;10(1):64-66
We present a patient who developed granuloma in a previous herpes zoster scar (post-zoster granuloma). The development of granuloma in healed herpes zoster lesions may represent an atypical delayed hypersensitivity reaction to viral antigens or tissue antigens altered by the virus. To our knowledge, this is the first case reported in Korean literature.
Antigens, Viral
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Cicatrix
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Granuloma*
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Herpes Zoster
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Humans
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Hypersensitivity, Delayed
4.Proliferation of Corneal Endothelial Cells by Delivery of SV40 Large T Antigen.
Kyung Sun CHO ; In Kyung SONG ; Timothy P FLEMING ; Choun Ki JOO
Journal of the Korean Ophthalmological Society 2001;42(9):1331-1335
PURPOSE: To determine whether the delivery of the SV40 large T-antigen is a feasible method for transiently inducing proliferation of corneal endothelial cells, we delivered liposome-protein complex into bovine corneal endothelial cells(BCEC). METHOD: SV40 large T-antigen protein was introduced into BCEC and positive cells were identified by immunohistochemistry. Quiescent BCECs were double-labeled using BrdU as a measure of de novo DNA synthesis and the Ki-67 was detected by standard immunohistochemical methods. RESULT: The treatment of quiescent BCECs with large T antigen caused an increase in BrdU incorporation and Ki-67 expression. It was tested by time-course study. CONCLUSION: This finding suggests that liposome-mediated delivery of transforming proteins could be a method to transiently induce corneal endothelial cell proliferation.
Antigens, Viral, Tumor*
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Bromodeoxyuridine
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Cell Proliferation
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DNA
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Endothelial Cells*
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Immunohistochemistry
5.Biological characteristics of influenza virus.
Chinese Journal of Pediatrics 2003;41(3):164-167
6.Transfection of SV 40 Large T Antigen into Corneal Endothelial Cells.
Kyung Sun CHO ; Timothy P FLEMING ; Eunjoo Hwang LEE ; Choun Ki JOO
Journal of the Korean Ophthalmological Society 1999;40(3):676-682
The coeneal endothelium is essential for the maintenance of normal corneal hydration, thickness, and transparency. However, corneal endothelial cells are incapable of significant proliferation in vivo. As we age, the density of corneal endothelial (CEN) cells gradually decreases. The goal of our study is to explore the possibility of enhancing the proliferation of corneal endothelial cells by introduction of SV 40 large T antigen, a transforming protein. To this end, introduction of protein into CEN cells was assessed by liposome assisted beta-galactosidase transfection in vivo, ex vivo, and in vivo. In all cases, cells treated with liposome-protein complex have shown dramatic blue stain in beta-galactosidase activity staining. This result convinced us that we could artificially introduce a foreign protein into a cell. To ascertain where SV 40 large T antigen is localized in the cell, purified SV 40 large T antigen was transfected into the cells using liposome and its presence was determined immunohistochemically. We show that the liposome delivered SV 40 large is localized in the nucleus and mitotic figures which may suggest its functional activity.
Antigens, Viral, Tumor*
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beta-Galactosidase
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Endothelial Cells*
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Endothelium
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Liposomes
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Transfection*
7.Polymerization and evaluation of the protective efficacy of rotavirus VP4* proteins.
Yijian LI ; Guoxing LUO ; Han YANG ; Lianzhi JIA ; Yuanjun ZENG ; Biyan ZHAO ; Tingdong LI ; Shengxiang GE
Chinese Journal of Biotechnology 2019;35(2):281-289
In previous studies, we found that truncated rotavirus VP4* (aa 26-476) could be expressed in soluble form in Escherichia coli and confer high protection against rotavirus in the mouse mode. In this study, we further improved the immunogenicity of VP4* by polymerization. The purified VP4* was polymerized through incubation at 37 ℃ for 24 h, and then the homogeneity of the particles was analyzed by HPLC, TEM and AUC, while the thermal stability and antigenicity was analyzed by DSC and ELISA, respectively. Finally, the immunogenicity and protective efficacy of the polymers analyzed by a mouse maternal antibody model. The results showed that VP4* aggregated into homogeneous polymers, with high thermostability and neutralizing antibody binding activity. In addition, VP4* polymers (endotoxin <20 EU/dose) stimulated higher neutralizing antibodies and confer higher protection against rotavirus-induced diarrhoea compared with the VP4* trimers when immunized with aluminium adjuvant. In summary, the study in VP4* polymers provides a new strategy for the development of recombinant rotavirus vaccines.
Animals
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Antibodies, Viral
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Antigens, Viral
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Capsid
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Capsid Proteins
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Mice
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Polymerization
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Rotavirus
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Rotavirus Infections
8.Qualitative detection of hepatitis C virus core antigen in the serum in patients with chronic hepatitis C.
Yi OUYANG ; De-ming TAN ; Tie-gang LI ; Hui ZHOU ; Chang TAN
Journal of Central South University(Medical Sciences) 2006;31(6):894-905
OBJECTIVE:
To evaluate the clinical significance of serum hepatitis C virus (HCV) core antigen detected by enzyme linked immunosorbent assay (ELISA).
METHODS:
The serum HCV core antigen, which was taken from 149 patients with chronic hepatitis C, 20 patients of chronic hepatitis B and 20 health volunteers, was detected by ELISA. Meanwhile, the serum HCV RNA was detected by RT-PCR, and anti-HCV was detected by ELISA.
RESULTS:
The qualitative HCV core antigen in the serum, which was take from 20 patients of chronic hepatitis B and 20 health volunteers, was negative.The positive percentage of HCV core antigen was 49.66% in the 149 sera of patients with chronic hepatitis C. The coincidence of detective results of HCV RNA and HCV core antigen was 54.36%, without significant difference (P>0.05). The positive percentage of HCV RNA and HCV core antigen in the 149 anti-HCV antibody positive sera samples were 55.03% (82/149) and 49.66% (74/149), respectively, and there was no significant difference (P>0.05).
CONCLUSION
The qualitative HCV core antigen detected by ELISA has a high specificity. The positive percentage of HCV core antigen in the serum of patients with chronic hepatitis C is 49.66%. HCV core antigen is related to HCV RNA. HCV core antigen may be a useful serum marker which could show HCV viraemia like HCV RNA.
Hepatitis C Antigens
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blood
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Hepatitis C, Chronic
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blood
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Humans
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RNA, Viral
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blood
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Viral Core Proteins
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blood
9.Expression of antigens of foot-and-mouth disease virus in plants: a review.
Yuting CAI ; Yi RU ; Kun SUN ; Ji ZHANG ; Jianping WU ; Dan LI ; Hanqing FENG
Chinese Journal of Biotechnology 2023;39(4):1548-1561
Foot-and-mouth disease (FMD) is an acute, severe, and highly contagious infectious disease caused by foot-and-mouth disease virus (FMDV), which seriously endangers the development of animal husbandry. The inactivated FMD vaccine is the main product for the prevention and control of FMD, which has been successfully applied to control the pandemic and outbreak of FMD. However, the inactivated FMD vaccine also has problems, such as the instability of antigen, the risk of spread of the virus due to incomplete inactivation during vaccine production, and the high cost of production. Compared with traditional microbial and animal bioreactors, production of antigens in plants through transgenic technology has some advantages including low cost, safety, convenience, and easy storage and transportation. Moreover, since antigens produced from plants can be directly used as edible vaccines, no complex processes of protein extraction and purification are required. But, there are some problems for the production of antigens in plants, which include low expression level and poor controllability. Thus, expressing the antigens of FMDV in plants may be an alternative mean for production of FMD vaccine, which has certain advantages but still need to be continuously optimized. Here we review the main strategies for expressing active proteins in plants, as well as the research progress on the expression of FMDV antigens in plants. We also discuss the current problems and challenges encountered, with the aim to facilitate related research.
Animals
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Foot-and-Mouth Disease Virus/genetics*
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Foot-and-Mouth Disease/prevention & control*
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Antigens, Viral/genetics*
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Viral Vaccines
10.Culture of HSV-2 and cloning of specific fragment of the gG-2 gene.
Jian GAO ; Yu-feng HUANG ; Jing CAO ; Hao-yang WANG ; Jin-chun LU
National Journal of Andrology 2009;15(3):223-227
OBJECTIVETo clone the glycoprotein G gene and its specific fragment with high conservation and antigenicity by culturing and amplifying herpes simplex virus type 2 and extracting its whole genome.
METHODSWe obtained a great deal of suspension with HSV-2 virus after infecting the cultured Hela cells with HSV-2 virus, extracted the whole genome of the virus by the phenol-chloroform method, and amplified the US4 gene coding gG-2 by PCR. Then we selected the specific target fragment according to the amino acid sequence alignment of the gG-2 gene and cloned it with the designed primers with restricted endonuclease sites.
RESULTSWe successfully obtained a lot of suspension with HSV-2 virus, and cloned the gG-2 gene from the whole genuine and its specific target fragment. Sequencing showed that both the sequences were identical with those printed in the GenBank.
CONCLUSIONIt is feasible to obtain the virus genome and specific fragment of the gG-2 gene from virus-infected cells, especially for HSV-2 virus with relatively stable hereditary trait. It has prepared the ground for further constructing the expression plasmid of the specific fragment, expressing related proteins and identifying their antigenicity.
Antigens, Viral ; genetics ; Cloning, Molecular ; DNA, Viral ; HeLa Cells ; Herpesvirus 2, Human ; genetics ; Humans ; Viral Envelope Proteins ; genetics ; Virus Cultivation