ORIGINAL_ARTICLE New insight into the application of outer membrane vesicles of Gram-negative bacteria This review presents a brief outline of our current knowledge of the structure and chemical composition of the outer membrane vesicles (OMVs), originating from the surface of Gram negative bacteria including their outer membrane proteins and lipopolysaccharides. Moreover, the functional roles and applications of OMVs in medical research such as OMV-based vaccines, OMV adjuvants properties, OMV carriers in conjugated vaccines as well as in drug vehicles and delivery systems are discussed. Finally, new applications of these macromolecules as biotechnological tools are briefly presented. http://vacres.pasteur.ac.ir/article-1-77-en.pdf 2016-07-25 1 4 10.18869/acadpub.vacres.2.5.93 OMV vaccine drug vehicles delivery systems. A Fateh afateh2@gmail.com 1 Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran, Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran AUTHOR F Vaziri farzam_vaziri@yahoo.com 2 Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran, Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran AUTHOR F Rahimi Janani 3 Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran, Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran AUTHOR S Ahmadi Badi 4 Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran AUTHOR M Ghazanfari 5 Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran AUTHOR M Davari 6 Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran AUTHOR A Arsang 7 Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran AUTHOR SD Siadat d.siadat@gmail.com 8 Mycobacteriology and Pulmonary Research Department, Pasteur Institute of Iran. Tehran, Iran, Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran AUTHOR
ORIGINAL_ARTICLE Synthesis and characterization of physicochemical and immunological properties of recombinant NS3-G2 dendrimer conjugate Introduction: An effective vaccine against HCV infection is not available. The non-structural protein 3 (NS3) of the virus as an important immunogenic candidate has been utilized in various modules. Nanostructured polymers have been recently used for efficient vaccine and drug delivery. The aim of the current study was the synthesis of rNS3-G2 conjugate and preliminary evaluation of its immunogenicity. Methods: The dendrimer was synthesized and conjugated with purified recombinant NS3 (rNS3) protein. The physicochemical properties of the conjugate were evaluated by Zeta potential, FT-IR spectra and confirmed by atomic force microscopy (AFM). Immunogenicity of the conjugate was assessed in BALB/c mice. Results: Synthesis and conjugation of dendrimer G2 with the protein were confirmed and immunological assays showed that the conjugated form of the antigen induced higher titer of IgG compared to rNS3 antigen alone. Conclusion: The results showed that the antigenic structure of rNS3 was maintained when conjugated with the biodegradable and biocompatible G2 dendrimers and the immunogenic properties of the antigen were enhanced. Therefore the new formulation may have potential as a vaccine candidate. http://vacres.pasteur.ac.ir/article-1-78-en.pdf 2016-08-14 5 8 10.18869/acadpub.vacres.2.5.97 Adjuvant Dendrimer G2 rNS3 HCV Conjugate F Javadi 1 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran. AUTHOR P Rahimi pooneh5376@yahoo.com, prahimi@pasteur.ac.ir 2 Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran. AUTHOR MH Modaressi modaresi@tums.ac.ir 3 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran. AUTHOR M Shafiee Ardestani shafieeardestani@gmail.com 4 Department of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran. AUTHOR A Bolhassani azam.bolhassani@yahoo.com 5 Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran. AUTHOR MR Aghasadeghi mr_sadeqi@yahoo.com 6 Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran. AUTHOR SM Sadat mehdi_sadat@yahoo.com 7 Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran. AUTHOR
ORIGINAL_ARTICLE Establishment of NS3 tumor cell line expressing Hepatitis C virus Non-structural Protein 3 as valuable tool for HCV challenging in mice Introduction: Hepatitis C virus (HCV) is one of the major medical problems. Human and chimpanzees are the only specific hosts which are naturally susceptible to HCV infection. Mice and other common laboratory animals are resistant to the virus, hence HCV prophylactic and therapeutic researches are very difficult and challenging. HCV non-structural protein 3 (NS3) is one of the most attractive targets for developing novel anti-HCV therapies as it is essential for the viral replication. This study was designed to produce stable SP2/0 tumor cell lines expressing NS3 of HCV for future basic and vaccine studies. Methods: A lentivector expressing NS3, named PCDH-NS3, was constructed by cloning of NS3 cDNA into downstream of CMV promoter of pCDH-CMV-MCS-EF1-Puro-GFP. The constructed plasmid was co-transfected with pMD2.G plasmid which encodes envelope VSV G protein and psPAX2 packaging plasmid into HEK-293T cells. The lentivector-containing supernatant was collected every 12 h for 72 h and NS3-Lentivector was concentrated by ultracentrifugation. Titers of the NS3 lentivector were estimated using flow cytometry. The SP2/0 cells were then infected by NS3 lentivector. Puromycin as a selective antibiotic was added to the culture for 2 weeks to select NS3 positive cells. A single transfected clone was obtained using limiting dilution. The 1st and 6th passages of the cells cultured in vitro were harvested and NS3 mRNA was detected for by RT-PC. Results: The results showed that NS3 expressing lentivector plasmids and the two other helper plasmids could be transfected into HEK293T efficiently and packaged successfully as a pseudo-lentivector. Finally, the detection of NS3 mRNA in the 1st and 6th passages of SP2/NS3 cells was confirmed by establishment of a stable cell line. Conclusion: SP2/0 Cell line with stable expression of NS3 can be used as a suitable tumor model to facilitate research on HCV vaccine in vitro and in mice model and it could be served as a valuable tool for pharmaceutical HCV research to pave the way for further research on NS3 vaccine function. http://vacres.pasteur.ac.ir/article-1-79-en.pdf 2016-08-17 9 14 10.18869/acadpub.vacres.2.5.101 HCV NS3 Lentivector Tumor challenging model. MH Pouriayevali mhpouriayevali@yahoo.com 1 Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran . AUTHOR T Bamdad bamdad_t@modares.ac.ir 2 Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran . AUTHOR SM Sadat mehdi_sadat@yahoo.com 3 Department of Hepatitis and AIDS, Pasteur Institute of Iran. Tehran, Iran. AUTHOR F Sabahi 4 Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran . AUTHOR MR Aghasadeghi mr_sadeqi@yahoo.com 5 Department of Hepatitis and AIDS, Pasteur Institute of Iran. Tehran, Iran. AUTHOR SH Yazdani yazdani.vrs2@gmail.com 6 Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. AUTHOR
ORIGINAL_ARTICLE Immunization of Litopenaeus vannamei shrimp against white spot syndrome virus (WSSV) by gamma-irradiated WSSV plus Vibrio paraheomolyticus Introduction: White spot syndrome virus (WSSV) is one of the most deadly infectious pathogens of the shrimp culture industry. Neither effective vaccines nor efficient treatments are currently available for this disease. Vibrio species are well known dominant bacterial pathogens in the shrimp ponds. As facultative pathogenic bacteria, it is possible that Vibrio spp. along with WSSV to co-infect the shrimp species such as Litopenaeus vannamei. The aim of this study was to investigate the co-administration of gamma-irradiated Vibrio paraheomolyticus as a kind of probiotic and immune-stimulator with gamma-irradiated and inactivated WSSV as an intramuscular vaccine for protection of L. vannamei against WSSV infection. Methods: WSSV was isolated from the infected shrimp samples and multiplied in Astacus leptodactylus crayfish. Titration of WSSV was obtained in post-larvae as 105.4 LD50/ml. The virus was irradiated where D10 value and optimum dose of gamma ray were calculated to be 2.56 and 15 kGy, respectively. The gamma-irradiated WSSV samples, named GI-WSSV were used as a vaccine to immunize L. vannamei shrimps. The freeze-dried V. paraheomolyticus cultures, inactivated by gamma ray (8 kGy) were named GI-V.P and used as a probiotic. Results: Protective dose50 was calculated as 5.61 and 7.94 for the shrimp groups which were vaccinated by GI–WSSV vaccine and GI-WSSV vaccine + GI-V.P, respectively. Calculated RPS values were 73.3%, 86.66% and 26.66% for the GI-WSSV vaccine, GI-WSSV + GI-V.P and isolated probiotic groups, respectively. Significant differences in cumulative mortalities were observed between the vaccination groups and the positive control group    (P < 0.05). No significant difference was observed in cumulative mortalities between the two vaccination groups        (P > 0.05). Conclusion: GI-WSSV vaccine can induce immune responses in shrimps infected with WSSV and probiotic GI-V.P enhances these responses. http://vacres.pasteur.ac.ir/article-1-80-en.pdf 2016-09-11 15 20 10.18869/acadpub.vacres.2.5.107 White Spot Syndrome Virus (WSSV) Shrimp Gamma Irradiation Vibrio paraheomolyticus Probiotic. F Motamedi-Sedeh fmotamedi@nrcam.org 1 Nuclear Science and Technology Research Institute, Tehran, Iran. AUTHOR M Afsharnasab 2 Iranian Fisheries Science Research Institute, Tehran, Iran. AUTHOR M Heidarieh 3 Nuclear Science and Technology Research Institute, Tehran, Iran. AUTHOR
SHORT_COMMUNICATION Purification and evaluation of glycerol teichoic acid from biofilm forming Staphylococcus epidermidis as a putative vaccine candidate Introduction: Biofilm forming Staphylococcus epidermidis is a main causative agent of infections related to medical devices. Purification and evaluation of Gly-TA polysaccharide from a biofilm-forming S. epidermidis as a putative vaccine candidate were the main goals of the current study. Methods: Taking advantage of size exclusion chromatography procedure, glycerol teichoic acid (Gly-TA) was purified from the above-mentioned strain and biochemical analyses including, Fourier Transform Infrared spectroscopy (FTIR) and Proton Nuclear Magnetic Resonance spectroscopy (H1-NMR) were conducted for the recovered polysaccharide. Results: Following PCR confirmation of a S. epidermidis strain, Gly-TA was extracted and its biochemical compositions (i.e. N-acetyl glucose amine residues) were obtained. Conclusion: It is envisaged that Gly-TA polysaccharide could be considered as a putative vaccine to inhibit formation of biofilm by S. epidermidis. http://vacres.pasteur.ac.ir/article-1-81-en.pdf 2016-09-21 21 25 10.18869/acadpub.vacres.2.5.113 Glycerol teichoic acid (Gly-TA) Staphylococcus epidermidis FTIR NMR. B Mirzaei dr.bahman.m@gmail.com 1 Department of Microbiology, Microbial research center, Pasteur Institute of Iran, Tehran, Iran, Microbiology Research Center (MRC) Pasteur Institute of Iran, Tehran, Iran. AUTHOR SF Moosavi 2 Department of Microbiology, Microbial research center, Pasteur Institute of Iran, Tehran, Iran, Microbiology Research Center (MRC) Pasteur Institute of Iran, Tehran, Iran. AUTHOR R Babaei 3 Microbiology Research Center (MRC) Pasteur Institute of Iran, Tehran, Iran, Department of Mycobacteriology and pulmonary research, Pasteur Institute of Iran, Tehran, Iran. AUTHOR SD Siadat d.siadat@gmail.com 4 Microbiology Research Center (MRC) Pasteur Institute of Iran, Tehran, Iran, Department of Mycobacteriology and pulmonary research, Pasteur Institute of Iran, Tehran, Iran. AUTHOR A Farjah 5 Young Researchers and Elite Club, East Tehran Branch, Azad University, Tehran, Iran. AUTHOR M Shahrooei 6 Laboratory of Medical Microbiology, Department of Medical Diagnostic Sciences, KU Leuven, U.Z. Gasthuisberg, Herestraat 49 CDG 8th floor, B-3000 Leuven, Belgium. AUTHOR
ORIGINAL_ARTICLE Differential protein expression in Mycobacterium tuberculosis susceptible and multidrug resistant isolates Introduction: Infections by multidrug resistant Mycobacterium tuberculosis (MDR-TB) is a major public health challenge. Secretion of proteins by M. tuberculosis plays an important role in the pathogenesis of the bacterium. We compared the protein profiles of susceptible M. tuberculosis and MDR-TB isolates using proteomic analyses, namely two dimensional gel electrophoresis (2DE) and mass spectrometry (MS). Methods: The bacilli were cultured on Middlebrook 7H9 medium and bacterial colonies were mechanically disrupted and proteins were extracted by ammonium sulfate. The 2DE and MS analyses were performed using Ettan IP Gphor 3 isoelecteric system and Autoflex II TOF/TOF, respectively. Results: Our study showed that in comparison to the sensitive strains, 27 proteins were over-expressed in the MDR isolates and these proteins were mainly involved in the cellular metabolism, cell wall and membrane structures and bacterial respiration. Bactoferritin (Rv1876) has been shown to play a role in antibacterial resistance. Increased intensity of Rv2031c, a heat shock protein (Alpha-crystallin/HspX), was also observed in the whole cell lysate of the MDR-TB. This protein is a marker of the latent TB and has been proposed as a target for vaccine development. Conclusion: Our results identified proteins that are overexpressed in the resistant M. tuberculosis which could be used as antibacterial targets or vaccine candidates.   http://vacres.pasteur.ac.ir/article-1-83-en.pdf 2016-09-28 26 30 10.18869/acadpub.vacres.2.5.118 Multidrug resistant Mycobacterium tuberculosis (MDR-TB) tuberculosis proteins protein candidate Mycobacteria antigen. AR Hadizadeh Tasbiti 1 AUTHOR Sh Yari tbchemistry@yahoo.com 2 AUTHOR M Ghanei 3 AUTHOR SD Siadat d.siadat@gmail.com 4 AUTHOR S Niknami 5 AUTHOR A Bahrmand padideh79@yahoo.com 6 AUTHOR
ORIGINAL_ARTICLE Bioinformatics study of complete amino acid sequences of neuraminidase (NA) antigen of H1N1 influenza viruses from 2006 to 2013 in Iran Introduction: Influenza is a contagious acute viral disease of the respiratory tract that causes fever, headache, muscle aches and cough. One of the unique features of influenza virus is antigenic variation in viral protein neuraminidase (NA) which causes emergence of new virus variants. NA is responsible for the release and spread of progeny virions. Due to the continuous changes of NA genes, vaccine strains must be re-selected annually. Methods: Complete NA amino acid sequences of 97 strains circulating from 2006 to 2013 in Iran were downloaded from NCBI. The sequences were edited and classified by the year of isolation and their diversity and important changes as well as changes in the predicted ligand binding sites and their resistance to anti-NA drugs, were analyzed. Bioinformatics software such as MEGA6.0, BioEdit, DNAsisMAX and DNAstar were used for the sequence alignments and phylogenetic analyses. Web-based analysis such as SWISS-MODEL, Phyre2 and 3DLigandSite were used for evaluation of the second and third protein structures and prediction of the ligand binding sites. Results: The results showed that 2009 could be considered as an important transition year which caused to classify the isolates into two different distinct groups. This shows the importance of changes made during possible mutations in the genomic structure of the virus which have made it antigenically different from the previous years. Anti-NA drug resistance was observed in 2009. This pandemic strain has become dominant in the following years and is used as a standard vaccine strain from 2010 onwards. Conclusion: The results obtained in this study can aid in better understanding of the antigenic evolution of H1N1 influenza viruses and can potentially accelerate the selection of the vaccine strains.   http://vacres.pasteur.ac.ir/article-1-84-en.pdf 2016-10-25 31 37 10.18869/acadpub.vacres.2.5.123 H1N1 influenza virus Neuraminidase Antigenic drift Sequence analysis Anti-NA drug resistance. A Farhangi ali54f@yahoo.com 1 Institute of Biochemistry and Biophysics, University of Tehran. Tehran. Iran. AUTHOR B Goliaei goliaei@ibb.ut.ac.ir 2 Institute of Biochemistry and Biophysics, University of Tehran. Tehran. Iran. AUTHOR K Kavousi 3 Institute of Biochemistry and Biophysics, University of Tehran. Tehran. Iran. AUTHOR A Ashtari 4 Razi Vaccine and Serum Research Institute, Karaj, Alborz, Iran. AUTHOR MA Bayatzadeh 5 Razi Vaccine and Serum Research Institute, Karaj, Alborz, Iran. AUTHOR A Pourbakhsh 6 Razi Vaccine and Serum Research Institute, Karaj, Alborz, Iran. AUTHOR
CASE_STUDY The Shift of HAV Infection in to adulthood in Iran: Consideration for HAV Vaccination http://vacres.pasteur.ac.ir/article-1-85-en.pdf 2016-10-25 38 38 10.18869/acadpub.vacres.2.5.130 Hepatitis A Vaccination Iran. MR Aghasadeghi mr_sadeqi@yahoo.com 1 Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran AUTHOR