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Alphaherpesvirus-vectored vaccines against animal diseases: Current progress
HU Yang, WANG Ming-shu, CHENG An-chun, JIA Ren-yong, YANG Qiao, WU Ying, LIU Ma-feng, ZHAO Xin-xin, ZHU De-kang, CHEN Shun, ZHANG Sha-qiu, WANG Yin, GAO Qun, OU Xu-min, MAO Sai, WEN Xing-jian, XU Zhi-wen, CHEN Zheng-li, ZHU Ling, LUO Qi-hui, TIAN Bin, PAN Lei-chang, Mujeeb Ur REHMAN, LIU Yun-ya, YU Yan-ling, ZHANG Ling, CHEN Xiao-yue
2020, 19 (8): 1928-1940.   DOI: 10.1016/S2095-3119(20)63175-6
Abstract242)      PDF in ScienceDirect      
Recombinant virus-vectored vaccines are novel agents that can effectively activate specific and nonspecific immunity, are multivalent and multieffective, and have high safety ratings.  Animal alphaherpesviruses have a large genome, contain multiple nonessential regions that do not affect viral replication and are capable of accepting the insertion of an exogenous gene and expressing the antigen protein.  Furthermore, animal alphaherpesviruses have a wide host spectrum, can replicate in the host and continuously stimulate the animal to produce immunity to the corresponding pathogen, thus making them ideal carriers for recombinant virus-vectored vaccines.  With the development of gene-editing technology, recombinant viruses capable of expressing foreign genes can be constructed by various methods.  Currently, studies on recombinant virus-vectored vaccines constructed based on animal alphaherpesviruses have involved poultry, pigs, cattle, sheep, and companion animals.  Studies have shown that the construction of recombinant animal alphaherpesviruses enables the acquisition of immunity to multiple diseases.  This article mainly summarizes the current progress on animal alphaherpesvirus-vectored vaccines, aiming to provide reference for the development of new animal alphaherpesvirus-vectored vaccines.
 
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Role of duck plague virus glycoprotein C in viral adsorption: Absence of specific interactions with cell surface heparan sulfate
JING Yan-chun, WU Ying, SUN Kun-feng, WANG Ming-shu, CHENG An-chun, CHEN Shun, JIA Ren-yong, ZHU De-kang, LIU Ma-feng, YANG Qiao, JING Bo, CHEN Xiao-yue
2017, 16 (05): 1145-1152.   DOI: 10.1016/S2095-3119(16)61550-2
Abstract931)      PDF in ScienceDirect      
Many mammalian herpes viruses utilize heparan sulfate (HS) moieties present on cell surface proteoglycans as receptors for cell entry, and this process also requires viral glycoprotein C (gC) homologues.  However, our understanding of the role of gC in facilitating attachment of other alpha-herpes viruses such as the duck plague virus (DPV) remains preliminary.  To study the role of gC during DPV infection, we used a gC-deleted mutant virus (DPV-ΔgC-EGFP).  Examination of the viral copy number by real-time PCR, as well as time course studies of viral adsorption and proliferation revealed that gC was involved in the viral binding to the cell surface.  The affinity of viral glycoproteins (gB-DPV, gC-DPV, and gE-DPV) to HS was assessed using a prokaryotic expression system and HiTrapTM Heparin HP column chromatography.  In addition, to confirm that gC played a role in the interaction between DPV and HS, viruses were treated with the HS analogue heparin and host cells were treated with its inhibitors heparinase prior to exposure to DPV-ΔgC-EGFP or wild-type strain Chinese virulent duck plague virus (DPV-CHv).  The effects of heparin and heparinase on virus infectivity demonstrated that function of gC on viral adsorption is independent of interactions between gC and heparin sulfate on cell surface.  All in all, this study demonstrated that the gC of DPV can mediate viral adsorption in an HS-independent manner, which distinguish it from the gC of some other alpha-herpes viruses.  Future studies will be required to identify the receptors involved in gC protein binding to cells.  This work provides us a foundation for further studies of examining the roles of gC in the adsorption during duck plague virus infection.
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Effects of antibacterial compounds produced by Saccharomyces cerevisiae in Koumiss on pathogenic Escherichia coli O8 and its cell surface characteristics
CHEN Yu-jie, WANG Chun-jie, HOU Wen-qian, WANG Xiao-shuo, GALI Bing-ga, HUASAI Si-mu-ji-de, YANG Si-qin, WU A-qi-ma, ZHAO Yu-fei, WU Ying-ga, CHEN Ao-ri-ge-le
2017, 16 (03): 742-748.   DOI: 10.1016/S2095-3119(16)61516-2
Abstract968)      PDF in ScienceDirect      
The effects of antibacterial compounds produced by Saccharomyces cerevisiae in Koumiss on pathogenic Escherichia coli O8 and its cell surface characteristics were investigated.  S. cerevisiae isolated from Koumiss produced antibacterial compounds which were active against pathogenic E. coli O8 as determined by the Oxford cup method.  The aqueous phases from S. cerevisiae at pH=2.0 (S2) and pH=8.0 (S8) were extracted and tested, respectively.  The organic acids of S2 and S8 were determined by high performance liquid chromatography (HPLC), and the concentrations of killer toxins were determined by enhanced bicinchoninic acid (BCA) Protein Assay Kit.  The minimum inhibition concentration (MIC) and the minimum bactericidal concentration (MBC) of S2 and S8 on E. coli O8 were determined by the broth microdilution method.  The effects of S2 and S8 on the growth curve of E. coli O8 were determined by turbidimetry, and the hydrophobicities of E. coli O8 cell surface were determined using the microbial adhesion to solvents method, the permeation of E. coli O8 cell membrane were determined by the o-nitrophenyl-β-D-galactoside (ONPG) method.  Aqueous phases at pH 2.0 and 8.0 had larger inhibition zones and then S2 and S8 were obtained by freeze-drying.  The main component in S2 was citric acid and it was propanoic acid in S8.  Other organic acids and killer toxins were also present.  Both the MICs of S2 and S8 on E. coli O8 were 0.025 g mL–1, the MBCs were 0.100 and 0.200 g mL–1, respectively.  The normal growth curve of E. coli O8 was S-shaped, however, it changed after addition of S2 and S8.  E. coli O8 was the basic character, and had a relatively hydrophilic surface.  The hydrophobicity of E. coli O8 cell surface and the permeation of E. coli O8 cell membrane were increased after adding S2 and S8.  The present study showed that S2 and S8 inhibit the growth of pathogenic E. coli O8 and influence its cell surface characteristics.
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Development and optimization of a double antibody sandwich ELISA for the detection of goose T cell surface CD8α molecule
ZHANG Wei, CHENG Bei-bei, CHEN Shun, WANG Ming-shu, JIA Ren-yong, ZHU De-kang, LIU Mafeng, LIU Fei, SUN Kun-feng, YANG Qiao, WU Ying, CHEN Xiao-yue, CHENG An-chun
2016, 15 (10): 2363-2368.   DOI: 10.1016/S2095-3119(16)61345-X
Abstract1649)      PDF in ScienceDirect      
    CD8, a glycoprotein on the surface of T cells, is involved in the defense against viral infection and plays significant roles in antigen presentation and in the antiviral immune response. CD8 is composed of two chains. Of these, the CD8α chain was chosen for the detection because it involved in both the CD8αα homodimer and the CD8αβ heterodimer. Here, we established a double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for specific detection of goose CD8α (goCD8α). The results showed that the optimal coated antibody and antigen dilutions were 1:50 (the antibody titer was 1:12 800) and 1:32 (0.3 ng mL–1), respectively, while the optimal capture antibody and horseradish peroxidase (HRP)-labelled goat anti-rabbit IgG dilutions were 1:50 (the antibody titer was 1:51 200) and 1:4 000 (the antibody titer was 1:5 000), respectively. The optimal blocking buffer was 5% bovine serum albumin (BSA). The best incubating condition was overnight at 4°C, the best blocking time was 120 min and the best anti-capture antibody working time was 150 min. In addition, the minimum dose detectable by DAS-ELISA was 5×10–3 ng mL–1. Most importantly, goCD8α expression levels in goose spleen mononuclear cells (MNCs) post-Goose parvoviruse (GPV) infection were found to be significantly up-regulated using the DAS-ELISA method, which was consistent with previous results obtained using real-time quantitative PCR. In conclusion, the DAS-ELISA method reported here is a novel, specific technique for the clinical detection of goCD8α.
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Identification of QTLs for grain size and characterization of the beneficial alleles of grain size genes in large grain rice variety BL129
GAO Xuan, ZHU Xu-dong, FANG Na, DUAN Peng-gen, WU Ying-bao, LUO Yue-hua, LI Yun-hai
2016, 15 (1): 1-9.   DOI: 10.1016/S2095-3119(15)61043-7
Abstract2422)      PDF in ScienceDirect      
Grain size is one of the most important agronomic components of grain yield. Grain length, width and thickness are controlled by multiple quantitative trait loci (QTLs). To understand genetic basis of large grain shape and explore the beneficial alleles for grain size improvement, we perform QTL analysis using an F2 population derived from a cross between the japonica variety Beilu 129 (BL129, wide and thick grain) and the elite indica variety Huazhan (HZ, narrow and long grain). A total number of eight major QTLs are detected on three different chromosomes. QTLs for grain width (qGW), grain thickness (qGT), brown grain width (qBGW), and brown grain thickness (qBGT) explained 77.67, 36.24, 89.63, and 39.41% of total phenotypic variation, respectively. The large grain rice variety BL129 possesses the beneficial alleles of GW2 and qSW5/ GW5, which have been known to control grain width and weight, indicating that the accumulation of the beneficial alleles causes large grain shape in BL129. Further results reveal that the rare gw2 allele from BL129 increases grain width, thickness and weight of the elite indica variety Huazhan, which is used as a parental line in hybrid rice breeding. Thus, our findings will help breeders to carry out molecular design breeding on rice grain size and shape.
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CDH1, a Novel Surface Marker of Spermatogonial Stem Cells in Sheep Testis
ZHANG Yan, WU Sachula, LUO Fen-hua, Baiyinbatu , LIU Lin-hong, HU Tian-yuan, YU Bo-yang, LI Guang-peng , WU Ying-ji
2014, 13 (8): 1759-1765.   DOI: 10.1016/S2095-3119(13)60689-9
Abstract1493)      PDF in ScienceDirect      
Spermatogonial stem cells (SSCs) are unique stem cells in adult body that can transmit genetic information to the next generation. They have self-renewal potential and can continuously support spermatogenesis throughout life of a male animal. However, the SSC population is extremely small, isolation and purification of the SSCs is challenging, especially for livestock animals. It has been confirmed that CDH1 (cadherin-1, also known as E-cadherin) can be expressed in undifferentiated SSCs of mouse and rats, but it has not been verified in sheep. Here, CDH1 was found as a novel surface marker for sheep SSCs. In this paper, sheep anti- CDH1 polyclonal antibodies were prepared and its activity was checked. Using the obtained antibodies and immunohistochemistry analysis, we confirmed that CDH1 can be expressed by SSCs in sheep testis.
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