1.Progress in research on molecular biology and application in dominant antigens ESAT6 and CFP10 of TB vaccine.
Journal of Biomedical Engineering 2012;29(2):392-396
As the dominant antigens, early secreted antigenic target 6 (ESAT6, E6) and culture filtrate protein 10 (CFP10, C10) had once been the focus of tuberculosis (TB) vaccine due to their capability of inducing strong cell immune response in the host. They are also endowed with promising future of prevention against and diagnosis of TB. In this review, we systematically introduce recent research progress of E6 and C10, especially in structure-function, biological characteristics, protein expression and secretion, host immunity and vaccine development, and the prospects of their application are also discussed.
Antigens, Bacterial
;
chemistry
;
genetics
;
immunology
;
Bacterial Proteins
;
chemistry
;
genetics
;
immunology
;
Humans
;
Immunodominant Epitopes
;
immunology
;
Molecular Biology
;
Peptide Fragments
;
chemistry
;
genetics
;
immunology
;
Tuberculosis Vaccines
;
genetics
;
immunology
;
Vaccines, DNA
;
immunology
2.Identification of Immunodominant B-cell Epitope Regions of Reticulocyte Binding Proteins in Plasmodium vivax by Protein Microarray Based Immunoscreening.
Jin Hee HAN ; Jian LI ; Bo WANG ; Seong Kyun LEE ; Myat Htut NYUNT ; Sunghun NA ; Jeong Hyun PARK ; Eun Taek HAN
The Korean Journal of Parasitology 2015;53(4):403-411
Plasmodium falciparum can invade all stages of red blood cells, while Plasmodium vivax can invade only reticulocytes. Although many P. vivax proteins have been discovered, their functions are largely unknown. Among them, P. vivax reticulocyte binding proteins (PvRBP1 and PvRBP2) recognize and bind to reticulocytes. Both proteins possess a C-terminal hydrophobic transmembrane domain, which drives adhesion to reticulocytes. PvRBP1 and PvRBP2 are large (> 326 kDa), which hinders identification of the functional domains. In this study, the complete genome information of the P. vivax RBP family was thoroughly analyzed using a prediction server with bioinformatics data to predict B-cell epitope domains. Eleven pvrbp family genes that included 2 pseudogenes and 9 full or partial length genes were selected and used to express recombinant proteins in a wheat germ cell-free system. The expressed proteins were used to evaluate the humoral immune response with vivax malaria patients and healthy individual serum samples by protein microarray. The recombinant fragments of 9 PvRBP proteins were successfully expressed; the soluble proteins ranged in molecular weight from 16 to 34 kDa. Evaluation of the humoral immune response to each recombinant PvRBP protein indicated a high antigenicity, with 38-88% sensitivity and 100% specificity. Of them, N-terminal parts of PvRBP2c (PVX_090325-1) and PvRBP2 like partial A (PVX_090330-1) elicited high antigenicity. In addition, the PvRBP2-like homologue B (PVX_116930) fragment was newly identified as high antigenicity and may be exploited as a potential antigenic candidate among the PvRBP family. The functional activity of the PvRBP family on merozoite invasion remains unknown.
Epitopes, B-Lymphocyte/*chemistry/genetics/*immunology
;
Female
;
Humans
;
Immunodominant Epitopes/chemistry/genetics/*immunology
;
Malaria, Vivax/immunology/*parasitology
;
Middle Aged
;
Plasmodium vivax/chemistry/genetics/*immunology
;
Protein Structure, Tertiary
;
Protozoan Proteins/chemistry/genetics/*immunology
;
Reticulocytes/*parasitology
3.Construction of an expression vector of GPC-3 CTL epitope.
Wen-jing WANG ; Cheng-yao LI ; Ming-song LI
Journal of Southern Medical University 2009;29(8):1548-1550
OBJECTIVETo construct the expression vectors of GPC-3 CTL epitope.
METHODSThe HBsAg gene with three different EYILSLEEL (EYI) sites was named EYI1, and another with one EYI replacing CTL epitope FLG or SIL of pcHBsAg were named EYI2 and EYI3, respectively. All the three DNAs were amplified by SEOing PCR from pcHBsAg plasmid and linked into pBSSK+ vector to construct Pbssk/EYI1, pBSSK/EYI2, and pBSSK/EYI3. The three plasmid were identified by PCR, double digestion and sequencing, and the fragments with EYI1-3 were obtained by double digestion and then inserted into pcDNA3.1+ vector.
RESULTS AND CONCLUSIONPCR, enzyme digestion and sequence analysis confirmed successful construction of the eukaryotic expression vectors pcDNA-EYI1/HBsAg, pcDNA-EYI2/HBsAg, pcDNA-EYI3/HBsAg, which facilitate further studies of the GPC3-HBsAg multiple peptides vaccine for HBV infection.
Amino Acid Sequence ; Epitopes ; chemistry ; genetics ; immunology ; Genetic Vectors ; genetics ; Glypicans ; genetics ; immunology ; metabolism ; Hepatitis B Surface Antigens ; genetics ; immunology ; Plasmids ; genetics ; metabolism ; Polymerase Chain Reaction ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccines, Subunit ; chemistry ; genetics ; immunology
4.Study on the B cell linear epitopes of rabies virus CVS-11 nucleoprotein.
Xin-Jun LV ; Xin-Xin SHEN ; Peng-Cheng YU ; Hao LI ; Li-Hua WANG ; Qing TANG ; Guo-Dong LIANG
Chinese Journal of Virology 2014;30(3):253-256
To study the B cell linear epitopes of rabies virus CVS-11 nucleoprotein, peptides were synthesized according to the amino acid sequences of B cell linear epitopes. Linear epitopes predicted by bioinformatics analysis were evaluated with immunological techniques. Indirect enzyme-linked immunosorbent assay showed that titers of antibodies to peptides (355-369 and 385-400 residues of rabies virus CVS-11 nucleoprotein) were above 1:12 800 in mouse sera. The antibodies recognized denatured rabies virus CVS-11 nucleoprotein in Western blot analysis. Purified anti-peptide antibodies recognized natural rabies virus CVS-11 nucleoprotein in BHK-21 cells in indirect fluorescent antibody test. The 355-369 and 385-400 residues of rabies virus CVS-11 nucleoprotein were validated as B cell linear epitopes.
Amino Acid Sequence
;
Animals
;
Antibodies, Viral
;
immunology
;
Epitope Mapping
;
Epitopes, B-Lymphocyte
;
chemistry
;
genetics
;
immunology
;
Female
;
Humans
;
Male
;
Mice
;
Mice, Inbred BALB C
;
Molecular Sequence Data
;
Nucleoproteins
;
chemistry
;
genetics
;
immunology
;
Rabies
;
immunology
;
virology
;
Rabies virus
;
chemistry
;
genetics
;
immunology
5.Removal of Alpha-Gal Epitopes from Porcine Aortic Valve and Pericardium using Recombinant Human Alpha Galactosidase A.
Seongsik PARK ; Woong Han KIM ; Sun Young CHOI ; Yong Jin KIM
Journal of Korean Medical Science 2009;24(6):1126-1131
It has been reported that the immune response due to alpha-Gal epitopes is an important factor in tissue valve failure. The elimination of the interaction between the natural anti-Gal antibodies and alpha-gal epitopes on the xenografts is a prerequisite to the success of xenografts in humans. Previously, we reported that the green coffee bean alpha-galactosidase could remove all alpha-Gal epitopes from cell surface of porcine aortic valve and pericardial tissue, but it has limitations on cost effectiveness. In this study we wanted to know whether the recently produced recombinant human alpha-galactosidase A has the same effective enzymatic activity as green coffee bean alpha-galactosidase in removing alpha-Gal epitopes from the same tissues. After treating fresh porcine aortic valve and pericardial tissue with recombinant alpha-galactosidase A, each sample was stained with Griffonia simplicifolia type I isolectin B4 indirect immunoperoxidase avidin-biotin technique. We then examined whether the alpha-Gal epitopes were reduced or abolished in each consecutive concentration of recombinant alpha-galactosidase A by comparing the degree of the Griffonia simplicifolia isolectin B4 staining. As a result, the recombinant alpha-galactosidase A could remove cell surface alpha-Gals on porcine aortic valve and pericardial tissue as effectively as green coffee bean alpha-galactosidase.
Adolescent
;
Animals
;
Aortic Valve/chemistry/cytology/*immunology
;
Child
;
Coffea/enzymology
;
Epitopes/*immunology
;
Heart Valve Prosthesis Implantation
;
Humans
;
Pericardium/chemistry/cytology/*immunology
;
Recombinant Proteins/genetics/*immunology
;
Swine
;
Transplantation, Heterologous/immunology
;
alpha-Galactosidase/genetics/*immunology
6.Identification of peptide mimotopes of an abroad-spectrum neutralizing epitope of highly pathogenic avian influenza hemagglutinin.
Hui-Juan SONG ; Wen-Xin LUO ; Zhen-Hua ZHENG ; Ying-Wei CHEN ; Yi-Xin CHEN ; Zi-Min CHEN ; Jun ZHANG ; Ning-Shao XIA
Chinese Journal of Virology 2008;24(6):421-426
A monoclonal antibody (8H5), which showed strong neutralization activity against 33 strains of H5N1 viruses isolated from hosts at various regions from 2002 to 2006, was characterized in our lab recently. This result indicated the presence of highly conserved neutralizing site on hemagglutinin (HA) of various H5N1 subtypes. In the present study, the peptide phage display technique was applied to generate mimotope of the conserved neutralizing epitope recognized by 8H5 mAb. Five peptides displayed on phage were identified to specifically bind to 8H5 mAb. One of the five peptides, 123, was further displayed on the virus-like particle assembled from aa 1-149 fragment of HBcAg. The chimeric particle HBc-T123 conserved the specific binding to 8H5 mAb, and competed with H5N1 viruses for 8H5 mAb. The antiserum induced by HBc-T123 intensively stained on SF21 cells infected by recombinant baculovirus containing HA gene of YU22 virus, indicating the production of cross-reactive antibody to H5N1 HA.
Amino Acid Sequence
;
Animals
;
Antibodies, Monoclonal
;
immunology
;
Epitopes
;
chemistry
;
genetics
;
immunology
;
Hemagglutinin Glycoproteins, Influenza Virus
;
chemistry
;
genetics
;
immunology
;
Humans
;
Influenza A Virus, H5N1 Subtype
;
chemistry
;
genetics
;
immunology
;
Influenza, Human
;
virology
;
Mice
;
Mice, Inbred BALB C
;
Molecular Sequence Data
;
Peptide Library
7.Mapping of the B Cell Neutralizing Epitopes on ED III of Envelope Protein from Dengue Virus.
Yaying LIN ; Kun WEN ; Yonghui GUO ; Liwen QIU ; Yuxian PAN ; Lan YU ; Biao DI ; Yue CHEN
Chinese Journal of Virology 2015;31(6):665-673
Dengue virus (DENV) envelope [E] protein is the major surface protein of the virions that indued neutralizing antibodies. The domain III of envelope protein (EDIII) is an immunogenic region that holds potential for the development of vaccines; however, the epitopes of DENV EDIII, especially neutralizing B-cell linear epitopes, have not been comprehensively mapped. We mapped neutralizing B-cell linear epitopes on DENV-1 EDIII using 27 monoclonal antibodies against DENV-1 EDIII proteins from mice immunized with the DENV-1 EDIII. Epitope recognition analysis was performed using two set of sequential overlapping peptides (16m and 12m) that spanned the entire EDIII protein from DENV-1, respectively. This strategy identified a DENV-1 type- specific and a group-specific neutralizing epitope, which were highly conserved among isolates of DENV-1 and the four DENV serotypes and located at two regions from DENV-1 E, namely amino acid residues 309-320 and 381-392(aa 309-320 and 381-392), respectively. aa310 -319(310KEVAETQHGT319)was similar among the four DENV serotypes and contact residues on aa 309 -320 from E protein were defined and found that substitution of residues E309 , V312, A313 and V320 in DENV-2, -3, -4 isolates were antigenically silent. We also identified a DENV-1 type-specific strain-restricted neutralizing epitope, which was located at the region from DENV-1 E, namely amino acid residues 329-348 . These novel type- and group-specific B-cell epitopes of DENV EDIII may aid help us elucidate the dengue pathogenesis and accelerate vaccine design.
Amino Acid Sequence
;
Animals
;
Antibodies, Neutralizing
;
immunology
;
Dengue
;
virology
;
Dengue Virus
;
chemistry
;
genetics
;
immunology
;
Epitope Mapping
;
Epitopes, B-Lymphocyte
;
chemistry
;
genetics
;
immunology
;
Humans
;
Mice
;
Molecular Sequence Data
;
Viral Envelope Proteins
;
chemistry
;
genetics
;
immunology
8.Molecular evolution of human influenza H3N2 virus hemagglutinin genes in Guangdong China.
Ping HUANG ; Jing ZHONG ; Li-Jun LIANG ; Nian-Mei HOU ; Han-Zhong NI ; Jie WU ; Xin ZHANG
Chinese Journal of Virology 2012;28(4):330-335
The molecular characterization and phylogenetic analysis of hemagglutinin (HA) genes of human influenza H3N2 viruses in Guangdong, China from 2007 to 2010 were studied in this study. By space-time sampling of strains, the HA genes of H3N2 strains from Guangdong were sequenced and searched from Internet, and then the variation and evolution of HA genes were conducted by Lasergene 7.1 and Mega 5.05 and evolutionary rates were analyzed by epidemiological data. The phylogenetic tree was established by alignment of 17 Guangdong strains and 26 global reference strains. Ks rates and Ka rates of HA genes were 2.06 x 10(-3)-2.23 x 10(-3) Nt/Year and 1.05 x 10(-3)-1.21 x 10(3) Nt/Year during 2007-2010, while the velocity of HA1 evolution of Ka was 3. 13 times than that of HA2 evolution. Compared with HA of vaccine strain A/Perth/16/2009, the genetic homologies of Guangdong strains in 2009 reached to 98.8%-99.7% and of Guangdong strains in 2010 reached to 98.0%-98.4%. There were some amino acid substitutions in five epitope regions of HA1 during 2007-2010, especially in B region (N160K) and D region (K174R/N); the K189E/N/Q and T228A in RBS (receptor-binding site) occurred in 2010 as two glycoproteins sites substituted impacted on the HA1 antigenicity. The antigenicity of epidemic H3N2 strains in 2010 was to some degree different that of the vaccine strain A/ Perth/16/2009. According to that there were variations of B and D epitopes and two sites of RBS and two glycoprotein in Guangdong H3N2 HA1 genes, WHO/ CDC should recommend new representative strains during 2011-2012 influenza seasons if H3N2 HA genes further evolve in the near future.
Amino Acid Substitution
;
China
;
Disulfides
;
chemistry
;
Epitopes
;
genetics
;
Evolution, Molecular
;
Hemagglutinin Glycoproteins, Influenza Virus
;
chemistry
;
genetics
;
immunology
;
Humans
;
Influenza A Virus, H3N2 Subtype
;
genetics
;
Mutation
;
Phylogeny
9.Reactivity of German Cockroach Allergen, Bla g 2, Peptide Fragments to IgE Antibodies in Patients' Sera.
Haeseok LEE ; Kyoung Yong JEONG ; Kwang Hyun SHIN ; Myung Hee YI ; Darambazar GANTULAGA ; Chein Soo HONG ; Tai Soon YONG
The Korean Journal of Parasitology 2008;46(4):243-246
Bla g 2 is a cockroach allergen of great importance. This study was conducted to identify IgE-binding epitope(s) of Bla g 2 using the recombinant protein technique. Approximately 50% of tested sera showed IgE reactivity to Pichiaexpressed Bla g 2 (PrBla g 2) and E. coli-expressed Bla g 2 (ErBla g 2). Only 5.3% of serum samples showed stronger reactivity to PrBla g 2 than ErBla g 2, indicating that serum was reactive to conformational or carbohydrate epitopes. The full-length and 5 peptide fragments of Bla g 2 were produced in E. coli. All fragments showed IgE-binding activity to the cockroach-allergy patients' sera. Specifically, peptide fragments of amino acid residue 1-75 and 146-225 appeared to be important for IgE-binding. The information about the IgE-binding epitope of Bla g 2 can aid in the diagnosis and treatment for cockroach allergies.
Adolescent
;
Adult
;
Aged
;
Amino Acid Sequence
;
Animals
;
Antibodies
;
Aspartic Endopeptidases/chemistry/genetics/*immunology
;
Child
;
Cockroaches/immunology
;
Epitopes
;
Gene Expression Regulation
;
Humans
;
Immunoglobulin E/*immunology
;
Middle Aged
;
Young Adult
10.Investigation of a special neutralizing epitope of HEV E2s.
Min YOU ; Lu XIN ; Yi YANG ; Xiao ZHANG ; Yingwei CHEN ; Hai YU ; Shaowei LI ; Jun ZHANG ; Zhiqiang AN ; Wenxin LUO ; Ningshao XIA
Protein & Cell 2014;5(12):950-953
Antibodies, Monoclonal
;
chemistry
;
immunology
;
Antigens, Viral
;
chemistry
;
genetics
;
immunology
;
Binding Sites
;
Capsid Proteins
;
chemistry
;
genetics
;
immunology
;
Epitopes
;
chemistry
;
genetics
;
immunology
;
Escherichia coli
;
genetics
;
metabolism
;
Gene Expression
;
Hepatitis E
;
immunology
;
prevention & control
;
virology
;
Hepatitis E virus
;
chemistry
;
immunology
;
Humans
;
Molecular Docking Simulation
;
Mutagenesis, Site-Directed
;
Peptide Mapping
;
Protein Binding
;
Recombinant Proteins
;
chemistry
;
genetics
;
immunology
;
Viral Hepatitis Vaccines
;
administration & dosage
;
biosynthesis