Publications by authors named "Bao-Ying Huang"

11 Publications

  • Page 1 of 1

Immunogenicity and safety of a severe acute respiratory syndrome coronavirus 2 inactivated vaccine in healthy adults: randomized, double-blind, and placebo-controlled phase 1 and phase 2 clinical trials.

Chin Med J (Engl) 2021 Apr 28;134(11):1289-1298. Epub 2021 Apr 28.

NHC Key Laboratory of Enteric Pathogenic Microbiology (Jiangsu Provincial Center for Disease Control and Prevention), Nanjing, Jiangsu 210009, China.

Background: The significant morbidity and mortality resulted from the infection of a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) call for urgent development of effective and safe vaccines. We report the immunogenicity and safety of an inactivated SARS-CoV-2 vaccine, KCONVAC, in healthy adults.

Methods: Phase 1 and phase 2 randomized, double-blind, and placebo-controlled trials of KCONVAC were conducted in healthy Chinese adults aged 18 to 59 years. The participants in the phase 1 trial were randomized to receive two doses, one each on Days 0 and 14, of either KCONVAC (5 or 10 μg/dose) or placebo. The participants in the phase 2 trial were randomized to receive either KCONVAC (at 5 or 10 μg/dose) or placebo on Days 0 and 14 (0/14 regimen) or Days 0 and 28 (0/28 regimen). In the phase 1 trial, the primary safety endpoint was the proportion of participants experiencing adverse reactions/events within 28 days following the administration of each dose. In the phase 2 trial, the primary immunogenicity endpoints were neutralization antibody seroconversion and titer and anti-receptor-binding domain immunoglobulin G seroconversion at 28 days after the second dose.

Results: In the phase 1 trial, 60 participants were enrolled and received at least one dose of 5-μg vaccine (n = 24), 10-μg vaccine (n = 24), or placebo (n = 12). In the phase 2 trial, 500 participants were enrolled and received at least one dose of 5-μg vaccine (n = 100 for 0/14 or 0/28 regimens), 10-μg vaccine (n = 100 for each regimen), or placebo (n = 50 for each regimen). In the phase 1 trial, 13 (54%), 11 (46%), and seven (7/12) participants reported at least one adverse event (AE) after receiving 5-, 10-μg vaccine, or placebo, respectively. In the phase 2 trial, 16 (16%), 19 (19%), and nine (18%) 0/14-regimen participants reported at least one AE after receiving 5-, 10-μg vaccine, or placebo, respectively. Similar AE incidences were observed in the three 0/28-regimen treatment groups. No AEs with an intensity of grade 3+ were reported, expect for one vaccine-unrelated serious AE (foot fracture) reported in the phase 1 trial. KCONVAC induced significant antibody responses; 0/28 regimen showed a higher immune responses than that did 0/14 regimen after receiving two vaccine doses.

Conclusions: Both doses of KCONVAC are well tolerated and able to induce robust immune responses in healthy adults. These results support testing 5-μg vaccine in the 0/28 regimen in an upcoming phase 3 efficacy trial.

Trial Registration: http://www.chictr.org.cn/index.aspx (No. ChiCTR2000038804, http://www.chictr.org.cn/showproj.aspx?proj=62350; No. ChiCTR2000039462, http://www.chictr.org.cn/showproj.aspx?proj=63353).
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http://dx.doi.org/10.1097/CM9.0000000000001573DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8183795PMC
April 2021

Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells.

Emerg Microbes Infect 2020 Dec;9(1):2105-2113

State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health and School of Life Sciences, Xiamen University, Xiamen, People's Republic of China.

The global pandemic of coronavirus disease 2019 (COVID-19) is a disaster for human society. A convenient and reliable neutralization assay is very important for the development of vaccines and novel drugs. In this study, a G protein-deficient vesicular stomatitis virus (VSVdG) bearing a truncated spike protein (S with C-terminal 18 amino acid truncation) was compared to that bearing the full-length spike protein of SARS-CoV-2 and showed much higher efficiency. A neutralization assay was established based on VSV-SARS-CoV-2-Sdel18 pseudovirus and hACE2-overexpressing BHK21 cells (BHK21-hACE2 cells). The experimental results can be obtained by automatically counting the number of EGFP-positive cells at 12 h after infection, making the assay convenient and high-throughput. The serum neutralizing titer measured by the VSV-SARS-CoV-2-Sdel18 pseudovirus assay has a good correlation with that measured by the wild type SARS-CoV-2 assay. Seven neutralizing monoclonal antibodies targeting the receptor binding domain (RBD) of the SARS-CoV-2 S protein were obtained. This efficient and reliable pseudovirus assay model could facilitate the development of new drugs and vaccines.
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http://dx.doi.org/10.1080/22221751.2020.1815589DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7534347PMC
December 2020

Novel heterozygous BPIFC variant in a Chinese pedigree with hereditary trichilemmal cysts.

Mol Genet Genomic Med 2019 06 29;7(6):e697. Epub 2019 Apr 29.

Department of Central Laboratory, Ningde Municipal Hospital, Fujian Medical University, Ningde, Fujian, China.

Background: Trichilemmal cysts (TCs) are common intradermal or subcutaneous cysts, which are commonly sporadic and rarely autosomal dominantly inherited. However, little is known about the disease-determining genes in families with TCs exhibiting Mendelian inheritance.

Objective: The aim of this study was to identify the causative gene in a family with TCs.

Methods: Whole-exome sequencing was performed on a TCs family to identify the candidate gene. Sanger sequencing was conducted to validate the candidate variants and familial segregation.

Results: We identified the heterozygous variant c.3G>C (p.Met1?) within the BPIFC gene. Sanger sequencing confirmed the cosegregation of this variant with the TCs phenotype in the family by demonstrating the presence of the heterozygous variant in all the 12 affected and absence in all the seven unaffected individuals. This variant was found to be absent in dbSNP141, 1,000 Genomes database and 500 ethnicity matched controls.

Conclusion: Our results imply that BPIFC is a causative gene in this Chinese family with hereditary TCs. Further studies should be performed to validate the role of BPIFC in the pathogenesis of this disease.
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http://dx.doi.org/10.1002/mgg3.697DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6565563PMC
June 2019

[The MicroRNA miR-205 inhibits epithelial-messenchymal transition in HK-2 cells by down-regulating ZEB1 and ZEB2 expressions].

Nan Fang Yi Ke Da Xue Xue Bao 2016 Dec;36(12):1700-1705

Central Laboratory, Ningde Municipal Hospital, Affiliated Hospital of Fujian Medical University, Ningde 352100, China. E-mail:

Objective: To explore the role of miR-205 in regulating epithelial-messenchymal transition (EMT) in proximal tubular cell line HK-2 cells and the underlying mechanism.

Methods: HK-2 cells transfected with miR-205 mimics or a scrambled control sequence were examined for miR-205 expressions and mRNA levels of ZEB1, E-cadherin, and α-SMA using real-time qPCR; the protein levels of ZEB1, ZEB2, E-cadherin, and α-SMA were detected with Western blotting. Immunohistochemistry was performed to examine the ectopic expression of β-catenin and E-cadherin expression in the cells.

Results: The expression levels of ZEB1 and ZEB2 decreased significantly (P<0.01) while E-cadherin expression was up-regulated (P<0.01) in cells transfected with miR-205 mimics. Transfection with miR-205 mimics also markedly down-regulated the expression of α-SMA (P<0.01), a marker of mesenchymal cells that play an important role in EMT of HK-2 cells. The ectopic expression of β-catenin was inhibited by miR-205 mimics in HK-2 cells.

Conclusion: miR-205 inhibits EMT in HK-2 cells by down-regulating the expression levels of ZEB1 and ZEB2.
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December 2016

[Effects of tanshinone IIA on Wnt/beta-catenin signaling pathway of high glucose induced renal tubular epithelial cell transdifferentiation].

Zhongguo Zhong Xi Yi Jie He Za Zhi 2012 Jul;32(7):965-9

The Affiliated Ning-de Municipal Hospital of Fujian University of Traditional Chinese Medicine, Fujian.

Objective: To observe the expressions of Wnt/beta-catenin and the effects of tanshinone IIA (TII A) on Wnt/beta-catenin signaling pathway in high glucose induced renal tubular epithelial cell transdifferentiation.

Methods: Human kidney proximal tubular epithelial cells (HK-2) were divided into three groups, i. e., the normal glucose group, the high glucose group, and the high glucose plus tanshinone IIA group. The expression of beta-catenin was observed using immunocytochemical staining. The protein expression of beta-catenin, E-cadherin, and alpha-smooth muscle actin (alpha-SMA) were detected by Western blot. The mRNA levels of beta-catenin and E-cadherin were detected by RT-PCR.

Results: Compared with the normal glucose group, both the protein and the mRNA expressions of beta-catenin were significantly enhanced (P < 0.01), the expression of E-cadherin significantly decreased (P < 0.01), the expression of beta-catenin increased in the cytoplasm and nucleus in the high glucose group. TIIA at the final concentration of 100 micromol/L significantly reduced the ectopic expression of beta-catenin. At that concentration, the protein and mRNA expressions of beta-catenin in the nucleus significantly decreased, while the protein and mRNA expressions of E-cadherin were up-regulated. Meanwhile, the expression of alpha-SMA obviously decreased.

Conclusions: Wnt/beta-catenin signaling pathway participated in the high glucose induced renal tubular epithelial cell transdifferentiation. TIIA inhibited the transdifferentiation process possibly through down-regulating the activities of Wnt/beta-catenin signaling pathway, thus further playing a role in renal protection.
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July 2012

[Characterization of hepatitis B virus antigen expression in vitro and in vivo transduced by different transfer plasmids carrying HBV infectious genome].

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi 2012 Apr;26(2):96-8

National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.

Objective: To character HBV antigen expression in vitro and in vivo transduced by different transgenic plasmids carrying infectious genome of hepatitis B virus (HBV).

Methods: We constructed four different lentiviral transfer plasmids (carrying 1.3 full-length genome of HBV, by replacing the EGFP express box in pCS-CG plasmid with HBV genome and with different structural element, named as pCS-HBV1.3 (pCS-HBV1.3 X, pCS-HBV1.3 P, pCS-HBV1.3 N and pCS-HBV1.3 K). We detected the expression of HBsAg and HBeAg by ELISA in different time after transfected Huh 7 cells or hydrodynamic injection into C57 BL/6 mice with transfer plasmids pCS-HBV, respectively.

Results: We detected significant expression of HBsAg over 5 days after transfected Huh 7 cells (in vitro) or hydrodynamic injection into C57 BL/6 mice (in vivo) with transfer plasmids pCS-HBV1.3 X, pCS-HBV1.3 P and pCS-HBV1.3 K. The expression level and dynamics of HBsAg and HBeAg in the sera of mice is consistent with that of in the supernatant of Huh-7 cell. Furthermore, the expression of HBV antigens were modulated by the direction and position of HBV insert, also by some lentiviral vector cis-elements (cPPT and RRE).

Conclusion: The optimal lentiviral transfer plasmids (pCS-HBV1.3 X, pCS-HBV1.3 P and pCS-HBV1.3 K) could be further used for establishment and application of HBV transgenic cell or animal model.
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April 2012

[The 50% effective dose (ED50a) of seasonal spilt influenza vaccine in mice].

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi 2011 Apr;25(2):92-5

National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.

Objective: To evaluate the seasonal influenza spilt vaccine's immunogenicity and the 50% effective dose (ED50a) of hemagglutin (HA) that can make 50% of the mice hemagglutination inhibition antibody (HI) titers to 40.

Methods: The 2008-2009 seasonal influenza spilt vaccine's two components, with HA from H1N1 and H3N2 influenza virus respectively, were used as a model. Mice were immunized once or twice with different doses, and the HI antibody titers were tested to determine the immunization procedure and to evaluate the immugenicity of seasonal influenza spilt vaccine in mice; Consequently, HI antibody response kinetics of the two components were observed to determine the time point when the HI antibody titer reached the peak point; Finally, mice were immunized with different doses of HA to evaluate the ED50a that can make 50% of mice HI titers reach 40.

Results: Immunization procedures study showed that one-dose of seasonal influenza vaccine induced the HI antibody titers ranged from 10 to 120, while two-dose of influenza vaccine improved the HI antibody titer 10-100 times as compared with one dose; antibody kinetics study suggested that the time point of HI antibody produced to peak is 28-35 days post one dose immunization; and the ED50a detection results indicated that one dose of 1.5 microg HA could make 50% of the mice HI antibody titer reach 40.

Conclusion: Seasonal influenza spilt vaccine is very immunogenic in mouse; the time point of HI antibody produced to peak is 28-35 days post one dose immunization; and the ED50a of HA is 1.5 microg, which can make 50% of the mice HI titer reach 40. The experimental results provided foundation for the establishment of influenza vaccine evaluation system based on seasonal influenza vaccine.
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April 2011

[Immune protective effectiveness of seasonal influenza spilt vaccine against homologous and heterogonous subtypes of influenza virus in mice].

Bing Du Xue Bao 2011 May;27(3):265-73

National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052 ,China.

To investigate the seasonal influenza split vaccine's immune protective effectiveness against the homologous and heterogonous subtypes of influenza A virus challenge and the relationship between the protective effectiveness and hemagglutination inhibition (HI) antibody titer in mice. Two components of H1N1 and H3N2 in Chinese 2008-2009 seasonal influenza spilt vaccine, were derived from vaccine strain A/Brisbane/59/2007 (H1N1)-like virus and A/Brisbane/10/2007 (H3N2)-like virus respectively, and were used to immune BALB/c mice. Firstly, different doses of the vaccines were used to immunize mice and the HA immunization dosage that can induce the HI antibody titer of 40 in mice was identified; Secondly, H1N1 vaccine immunized mice were challenged with different doses of influenza virus mouse adaptation strains of A/Brisbane/59/2007 (H1N1)-like virus (MA) (referred to as A1 virus, well matched-strain in the homologous subtype) and A/Purto Rico/8/34 (H1N1) (referred to as PR8 virus, poor matched-strain in the homologous subtype) respectively, and H3N2 vaccine immunized mice were challenged with H1N1 influenza virus of A1 strain (Heterogonous subtype), body weight changes and survival rates were observed to explore the immune protective effectiveness of influenza split vaccine against the homologous and heterogonous subtypes of influenza A virus in mice. Results indicated that HI antibody titers were elevated as the HA protein immunization dosages increased from 0.15 microg, 0.5 microg, 1.5 microg, 5 microg to 15 microg in mice, and 1.5 microg HA of the seasonal influenza split vaccine could induced HI antibody titer of 40 in mice; 3LD50, 10LD50, 30LD50, 100LD50, 300LD50,1000LD50 and 3000LD50 of influenza virus strain A1 were used to challenge the H1N1 immunization mice, 1.5 microg HA of H1N1 vaccine could 100% protect mice against challenge with 1000LD50 of matched and homologous subtype of influenza virus strains A1, mice immunized with 15 microg HA of H1N1 vaccine even could 100% protect mice against challenge with 3000LD50 of influenza virus strains A1; but mice immunized with both the 1.5 microg and 15 microg HA of H1N1 vaccine were all sacrificed when challenged with 3LD50 of the mismatched and homologous subtype of influenza virus strain PR8, and mice immunized with the high dosage of 15 microg HA of H3N2 vaccine also were all sacrificed when challenged with 3LD50 of the heterogonous subtype of influenza virus strain A1. These results suggest that 1.5 microg HA of seasonal influenza split vaccine could induced HI antibody titer of 40 after one dose in mice, this dosage of HA can effectively protect mice against matched homologous subtype of influenza virus strain, but hardly to protect mice against mismatched homologous or heterogonous subtype of influenza virus strain. These results provide materials for the establishment of influenza vaccine evaluation system based on seasonal influenza vaccine.
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May 2011

[Efficient soluble expression and purification of influenza A nucleoprotein in Escherichia coli].

Bing Du Xue Bao 2011 Jan;27(1):50-7

National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.

To efficiently express nucleoprotein (NP) of influenza A virus A/Jingke/30/95 (H3N2) in E. coli for further immunogenicity study, three forms of NP gene, NP(His) (NP fused with 6 x His tag), NPwt (wild type NP, non-fused NP with native codon) and NP(O) (codon optimized, non-fused NP) were cloned by the technologies of restriction enzyme digestion, PCR, codon optimization and gene synthesis. Three recombinant plasmids were subsequently constructed based on the prokaryotic vector pET-30a, respectively. The comparative studies with these plasmids were carried out on the gene expression efficiency, induction temperature and time, purification process and immune reactivity. It was confirmed by restriction enzyme digestion and sequencing analysis that the three NP genes were inserted into the expression plasmid pET-30a correctly. SDS-PAGE showed that all three forms of NP gene could be efficiently ex pressed in E. coli, among which NP(O) was expressed with the highest expression level. The lower temperature fermentation (T=25 degrees C) and longer time induction (t=10 h) were necessary for high-level expression of protein in soluble form. The purity of tag-free NP was up to 90% through the two-step purification process with anion-exchange and gel filtration chromatography. It was indicated by Western blot that purified NP reacted well with the serum from mice immunized with PR8 virus. These results suggest that the codon-optimized influenza A virus NP gene can be efficiently expressed in E. coli and the expressed NP protein with specific immune reactivity could be purified from the supernatant of bacterial lysate.
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January 2011

[Expression of influenza A3 virus (H3N2) M2 gene in vaccinia virus Tiantan strain].

Bing Du Xue Bao 2007 Sep;23(5):377-83

Biotech Center for Viral Disease Emergency, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.

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.
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September 2007