@article{ author = {Taheri, Tahereh and Gholami, Elham and Saatchi, Faeze and Seyed, Negar and Taslimi, Yasaman and Rafati, Sim}, title = {Expressional Comparison Between Episomal and Stable Transfection of a Selected tri-fusion Protein in Leishmania tarentolae}, abstract ={  Introduction: Leishmania tarentolae (L. tarentolae) is a nonpathogenic species of Leishmania genus that can be used as an expression system to produce immunogenic proteins or epitopes in vivo acting as an efficient and safe recombinant live vector in vaccinology. Cysteine proteases (CPs) are one group of Family C1 peptidases in Leishmania that are important for survival of the parasite within the host and are introduced as potential vaccine candidates. Two types of cysteine proteases, CPA and CPB, play a critical role in Leishmania. Previously, it has indicated that immunization with two genes or recombinant proteins of CPA and CPB individually or fused together with various adjuvant or combined with liposome are able to elicit a protective immune response against L. major in BALB/c mice. Methods: In this study, for the first time, two lines of recombinant non-pathogenic Leishmania were generated by transfection of a heterologous triple fusion gene encoding cpa/cpb/egfp. For this purpose, two different expressions approaches were taken episomal expression using rDNA promoter, and integrative expression from rRNA locus of genome. Results: After genotype confirmation, the fluorescence intensity was monitored in different phases of parasite growth by both fluorescence microscopy and FACS analysis.Western blot and RT-PCR analysis showed that cpa/cpb/egfp tri-fused genes were specifically expressed in both lines of transgenic parasites. Discussion: In this study a single fused gene was expressed in both systems and the results showed that the expression of integrated gene was higher than episomal. In addition, the present data suggest that L. tarentolae expressing cpa/cpb/egfp is a promising carrier as vectored vaccine in future research. Vac Res , 2014, 1 (1): 1-9}, Keywords = {Leishmania tarentolae, Knock-in, Cysteine Protease, EGFP}, volume = {1}, Number = {1}, pages = {1-9}, publisher = {Pasteur Institute of Iran}, title_fa = {}, abstract_fa ={}, keywords_fa = {}, doi = {10.18869/acadpub.vacres.1.1.1}, url = {http://vacres.pasteur.ac.ir/article-1-33-en.html}, eprint = {http://vacres.pasteur.ac.ir/article-1-33-en.pdf}, journal = {Vaccine Research}, issn = {2383-2819}, eissn = {2423-4923}, year = {2014} } @article{ author = {Arabi, Samira and Aghasadeghi, Mohammad Reza and Memarnejadian, Arash and Kohram, Fatemeh and Aghababa, Haniyeh and Khoramabadi, Nima and Taghizadeh, Morteza and Shahosseini, Zahra and Mahdavi, Mehdi}, title = {Cloning, Expression and Purification of a Novel Multi-epitopic HIV-1 Vaccine Candidate: A Preliminary Study on Immunoreactivity}, abstract ={  Introduction : Designing an effective vaccine against human immunodeficiency virus (HIV)-1 is a global health priority . Multi-epitope vaccines offer several potential advantages that may be promising in case of mutable divergent pathogens such as HIV-1. Herein, a multiepitopic recombinant protein containing various HIV-1 antigens was expressed in E. coli cells and its immunogenicity in combination with different adjuvants was initially evaluated in BALB/c mouse. Methods: HIVtop4 sequence spanning the junction of six amino acid fragments (Gag158-186, Pol150-190, ENV296-323, ENV577-610, Tat1-20 and Tat44-61) was designed based on immunoinformatic analysis to reduce the creation of junctional epitopes, improve the cleavage of proteasome and avoid the local accumulation of hydrophobic regions. Synthesized nucleotide sequence corresponding to HIVtop4 was cloned into pET23a plasmid. Expression of pET-HIVtop4 plasmid was induced in BL21 (DE3) E. coli cells by addition of 1 mM IPTG during 3 h culture and the protein was purified by Ni-NTA column chromatography and further confirmed against anti-His antibody in western-blotting. Groups of BALB/c mice (n=6) were immunized three times with 2 weeks interval, subcutaneously with 10 m g of candidate vaccine adjuvanted in Complete Freund’s adjuvant , Montanide ISA70 and Alum with suitable control groups. Two weeks after last immunization lymphocyte proliferation was measured with Brdu, IL-4 and IFN- g cytokines with ELISA, total antibody and IgG1, IgG2a isotypes with indirect ELISA methods. Results: Results showed that Immunization with HIV-1 tat/pol/gag/env led to a significant increase in the proliferative responses of lymphocytes, IL-4 and IFN-γ cytokine production and humoral immune response in comparison with the control groups. Conclusion: In this study we concluded that Tat, Env, Pol, Gag with adjuvants (Montanide, Alum and CFA) has potentials as a candidate vaccine against the HIV-1 virus. Vac Res, 2014, 1 (1): 10-15}, Keywords = {HIV-1 tat/pol/gag/env, multi-epitope, Protein expression, Immune response}, volume = {1}, Number = {1}, pages = {10-15}, publisher = {Pasteur Institute of Iran}, title_fa = {}, abstract_fa ={}, keywords_fa = {}, doi = {10.18869/acadpub.vacres.1.1.10}, url = {http://vacres.pasteur.ac.ir/article-1-27-en.html}, eprint = {http://vacres.pasteur.ac.ir/article-1-27-en.pdf}, journal = {Vaccine Research}, issn = {2383-2819}, eissn = {2423-4923}, year = {2014} } @article{ author = {Golshani, Maryam and Zandi, Payam and Bouzari, Saei}, title = {In silico Design of Truncated Omp31 Protein of Brucella melitensis: Its Cloning and High Level Expression in Escherichia coli}, abstract ={  Introduction: Omp31 is animmunodominant and protective antigen conserved in Brucella species and a good candidate for vaccine design. Material & methods: The present study aimed at in silico design of the truncated Omp31 (TOmp31) using bioinformatic tools and to express the selected form in Escherichia coli (E. coli) Results and conclusion: Various bioinformatically calculated scores for the model showed that the structure conformation of the truncated Omp31 is in the range of the native protein with the C-score, Z-score, TM-score and Ramachandran Z-score of the truncated form model being -0.53, -0.72 and -0.98 respectively. Amplification of TOmp31 produced a single fragment of approximately 345 bp which was cloned in the pET28a expression vector. The integrity of the constructed vector (pET28- TOmp31) was confirmed by PCR and restriction digestion analysis and the rTOmp31 was successfully expressed in E. coli BL21. SDS-PAGE analysis of the lysate from the induced E. coli carrying the TOmp31 construct and the purified protein showed the expected molecular mass of approximately 12 kDa. The yield of the purified protein was estimated at approximately 250 μg/ml. Anti-His antibody reacted with the purified protein in Western blot confirming its expression in the prokaryotic system. Future studies exploring the immunogenicity and cross-protection of the protein against Brucella spp. are underway. Vac Res , 2014, 1 (1): 16-21}, Keywords = {Brucella, Omp31, Modeling, Truncated, In silico analysis}, volume = {1}, Number = {1}, pages = {16-20}, publisher = {Pasteur Institute of Iran}, title_fa = {}, abstract_fa ={}, keywords_fa = {}, doi = {10.18869/acadpub.vacres.1.1.16}, url = {http://vacres.pasteur.ac.ir/article-1-31-en.html}, eprint = {http://vacres.pasteur.ac.ir/article-1-31-en.pdf}, journal = {Vaccine Research}, issn = {2383-2819}, eissn = {2423-4923}, year = {2014} } @article{ author = {Daemi, Amin and Hosseinzadeh, Sahar and Bolhassani, Azam and Agi, Elnaz}, title = {DNA-Based Vaccine Is More Efficient than Non-Pathogenic Live Vaccine for the Prevention of HPV16 E7-Overexpressing Cancers}, abstract ={  Introduction: Vaccinology provides promising approaches for the control of various infectious diseases. Among different strategies, DNA vaccines offer attractive research opportunities for development of vaccines for induction of antigen-specific immunity owing to their stability, simplicity of delivery, safety and cost effectiveness. However, there is a need to increase their potency by the use of adjuvants such as glycoprotein 96 and electroporation delivery. On the other hand, the attenuated or non-pathogenic live vectors have been used to deliver DNA into cells as efficient delivery tools in gene therapy. Recently, a non-pathogenic protozoan, Leishmania tarentolae (L. tar), has attracted attention as an in situ protein-delivery vehicle. Cervical cancer is the second largest cause of cancer deaths among women worldwide and human papillomaviruses (HPV) is reportedly a frequent cause of this type of cancer. Methods: In the current study, we compared the potential of live L. tar-based and DNA-based vaccines expressing HPV16 E7 linked to C-terminal fragment of gp96 in a tumor mouse model. Results: We found that subcutaneous DNA injection with E7-CT (gp96), followed by electroporation, generate a significant E7-specific IFN-γ immune response and in vivo protective effects compared to transgenic L. tar-E7-CT (gp96) in challenge experiments with TC-1. Conclusion: It could be concluded that the DNA vaccine showed higher efficacy compared to the non-pathogenic live parasite-based vaccine in the tumor mice model. Vac Res , 2014, 1 (1): 22-25}, Keywords = {DNA vaccine, Live vaccine, Leishmania tarentolae, Glycoprotein 96, Electroporation}, volume = {1}, Number = {1}, pages = {21-24}, publisher = {Pasteur Institute of Iran}, title_fa = {}, abstract_fa ={}, keywords_fa = {}, doi = {10.18869/acadpub.vacres.1.1.21}, url = {http://vacres.pasteur.ac.ir/article-1-30-en.html}, eprint = {http://vacres.pasteur.ac.ir/article-1-30-en.pdf}, journal = {Vaccine Research}, issn = {2383-2819}, eissn = {2423-4923}, year = {2014} } @article{ author = {Rahimi, Pooneh and Shirzadi, Mohammad Reza and Farahtaj, Firouzeh and Fallahian, Vida and Sharifian, Jamal and Howaizi, Nader and Shamsipour, Mansour and Vahabpour, Rouhollah}, title = {Efficacy of Purified Vero Cell Rabies Vaccine (PVRV) under the Zagreb Regimen in Iran}, abstract ={  Background: Despite the effective pre- and post-exposure treatments, at least 60,000 deaths from rabies occur worldwide every year. Post-exposure treatment is considered as one of the most significant measures for preventing human deaths in exposed individuals. The 2-1-1 rabies post-exposure treatment schedule known as Zagreb regimen is an abbreviated immunization plan in which a tissue culture rabies vaccine is administered intramuscularly at two sites on day 0 and at one site on days 7 and 21. Objectives: In this study the efficacy of rabies vaccine administration under Zagreb regimen in an Iranian group of patients attending Pasteur Institute of Iran was evaluated. Methods: Rabies neutralizing antibody titer was measured in 75 serum samples collected from 25 volunteers receiving post exposure treatment by rapid fluorescent focus inhibition test (RFFIT), and ELISA. Results: All patients were negative for rabies antibody in both ELISA and FRRIT tests on day 0 . A satisfactory rabies virus antibody response with the titer of ≥0.5 IU/ml was detected in all patients on day 21 and two weeks after the completion of vaccination (day 35). Conclusions: Rabies immunization with rabies Vero-cell vaccine (PVRV) under the 2-1-1 schedule (Zagreb regimen) could result in an adequate immune response without any adverse effect. However a more comprehensive comparative study is underway to confirm these findings . Vac Res , 2014, 1 (1): 26-28}, Keywords = {Zagreb regimen, PVRV, Rabies vaccines, Post-exposure prophylaxis}, volume = {1}, Number = {1}, pages = {25-27}, publisher = {Pasteur Institute of Iran}, title_fa = {}, abstract_fa ={}, keywords_fa = {}, doi = {10.18869/acadpub.vacres.1.1.25}, url = {http://vacres.pasteur.ac.ir/article-1-32-en.html}, eprint = {http://vacres.pasteur.ac.ir/article-1-32-en.pdf}, journal = {Vaccine Research}, issn = {2383-2819}, eissn = {2423-4923}, year = {2014} } @article{ author = {Arsang, Amin and Yari, Shamsi and Masoumi, Morteza and NourNeamatollahi, Ali and Vaziri, Farzam and Nejati, Mehdi and Bahremand, Ahmad Reza and Siadat, Seyed Davar}, title = {Extraction and Purification of Haemophilus influenzae Type b Lipooligosac‌charide by Modified Phenol Method}, abstract ={  Introduction : Haemophilus influenzae type b (Hib) is a Gram negative bacterium and one of the causative agents of acute bacterial meningitis, especially in infants and children less than 5 years old. Lipooligosaccharide (LOS), one of the virulence factors which plays an important role in pathogenesis of Hib, has multiple applications in diagnosis and conjugate vaccines. In this study, LOS extracted from two Hib standard strains (ATCC 39930 and ATCC 10214) were compared. Material and methods: LOS was extracted by a modified hot phenol method from the aqueous and phenol phase and its concentration and purity assayed . Protein c ontaminations of the samples were determined spectrophotometrically and their endotoxin contents were assayed by the Limulus amebocyte lysate (LAL). Result and Discussion: The yield of the extracted LOS from strain ATCC10214 was about 475 μg/ml and the protein contaminations of the samples were approximately 0.07 mg/ml, whereas strain ATCC39930 yielded 520 μg/ml of LOS with protein contamination of 0.08 mg/ml. The results showed that the production of LOS by both strains was similar and the variation observed was not statistically significant (p < 0.001). Vac Res , 2014, 1 (1): 29-31}, Keywords = {Haemophilus influenzae type b, Lipooligosaccharide, Meningitis}, volume = {1}, Number = {1}, pages = {28-30}, publisher = {Pasteur Institute of Iran}, title_fa = {}, abstract_fa ={}, keywords_fa = {}, doi = {10.18869/acadpub.vacres.1.1.28}, url = {http://vacres.pasteur.ac.ir/article-1-28-en.html}, eprint = {http://vacres.pasteur.ac.ir/article-1-28-en.pdf}, journal = {Vaccine Research}, issn = {2383-2819}, eissn = {2423-4923}, year = {2014} } @article{ author = {Asgary, Vahid and KordMafi, Omid and Khosravy, Mohammad Sadeq and Janani, Alireza and NamvarAsl, Nabiollah and Bashar, Rouzbeh and Poortaghi, Hadi and AhangariCohan, Hossein and Shoari, Alireza and AhangariCohan, Rez}, title = {Evaluation of the Effect of Silver Nanoparticles on Induction of Neutraliz-ing Antibodies against Inactivated Rabies Virus}, abstract ={  Background: Nanoparticles have been considered as promising tools because of their high applicability. Recently, nanoparticles have been evaluated for their ability to increase the immune responses as adjuvants. Silver-nanoparticles (AgNPs) have shown promising results in enhancing Th-2 immune responses and to produce potent neutralizing antibodies. Neutralizing antibodies are considered as the main defense mechanism in pre- and post-exposure treatments of rabies disease. Therefore in this study, the effects of AgNPs in enhancing the immunogenicity of inactivated rabies virus were assessed. Materials and Methods: Different concentrations of AgNPs (0.2, 0.4, 0.6 and 0.8 mg/ml) were added to inactivated rabies virus. Mice were immunized by two intra-peritoneal injections of each concentration on days 1 and 7. The inactivated virus and Alum were used as negative and positive controls, respectively. Blood was collected from healthy and immunized mice, one week after the last immunization. Serum was isolated from each sample and the amounts of neutralizing antibodies were determined by Rapid Florence Focus Inhibition Test (RFFIT). The cytotoxicity of AgNPs was also assessed by in vitro MTT assay on J774A.1 cell line. Results: The results showed that 0.4, 0.6 and 0.8 mg/ml of AgNPs had significantly increased the immune responses compared to the control however, 0.2 mg/ml of AgNPs did not show a significant effect. No cytotoxicity was observed for 0.001 and 0.01 mg/ml concentrations of AgNPs but cell viability was decreased significantly at 0.1 mg/ml concentration. Conclusion: It was shown that the virus-loaded AgNPsat 0.4 mg/ml concentration could raise the neutralizing antibodies against rabies virus in mice, but their adverse effect on cell viability excludes their use as an adjuvant. Vac Res , 2014, 1 (1): 32-35 }, Keywords = {Adjuvant, Silver Nanoparticle, Rabies Vaccine, Neutralizing Antibodies}, volume = {1}, Number = {1}, pages = {31-34}, publisher = {Pasteur Institute of Iran}, title_fa = {}, abstract_fa ={}, keywords_fa = {}, doi = {10.18869/acadpub.vacres.1.1.31}, url = {http://vacres.pasteur.ac.ir/article-1-29-en.html}, eprint = {http://vacres.pasteur.ac.ir/article-1-29-en.pdf}, journal = {Vaccine Research}, issn = {2383-2819}, eissn = {2423-4923}, year = {2014} } @article{ author = {Abdoli, Asghar and Soleimanjahi, Hoorieh and Jamali, Abbas and Gholami, Shima and AminiRissehei, Abdolhossein and NamvarAsl, Nabiollah and Biglari, Peyvand and TavassotiKheiri, Masoumeh}, title = {Optimization of Microcarrier-based MDCK-SIAT1 Culture System for Influenza Virus Propagation}, abstract ={  Introduction: The preparation of seasonal influenza virus vaccines and especially its large-scale production requirement after the emergence or reemergence of a pandemic will need an alternative host cell system due to current suboptimal methods and the insufficiency of embryonated chicken eggs needed for producing them. In response to the vital and increasing demand for alternative means for influenza vaccine production, a cell line culture on microcarriers could be a potential alternative to the egg-based production. Methods: Influenza A/PR/8/1934 H1N1 was purified and quantified by plaque assay. The purified virus with 0.01 multiplicity of infection (MOI) was inoculated on Madin-Darby canine kidney-Siat1 (MDCK-SIAT1) cell line. Cytodex-1 microcarrier beads (2 g/l and 2.0×105cells/ml) were used in a spinner flask to culture MDCK-SIAT1cells .The culture medium was harvested and clarified and the virus yield was quantified by 50% cell culture infective dose ( CCID50) and hemagglutination assays. Next, the virus was concentrated and purified by ultra-filtration and ultra-centrifugation, respectively. Results: MDCK-SIAT1cells attached to the microcarriers and the cell numbers were increased efficiently. The cellular yield from the microcarrier culture was 2×106 cells/ml after 4-5 days. The yield of the virus titer measured by CCID50 and hemagglutination assays after the clarification was 108 CCID50/ml and 40960 HA unit/ml, respectively. Conclusion: MDCK-SIAT1cells may be considered as a new substrate for the production of influenza vaccines. Using Cytodex-1 microcarrier beads can be an appropriate strategy to improve the viral yield and to lower the cost of influenza vaccine production. Vac Res , 2014, 1 (1): 36-40}, Keywords = {Influenza virus, Cytodex 1, Vaccine, MDCK-SIAT1 cells}, volume = {1}, Number = {1}, pages = {35-38}, publisher = {Pasteur Institute of Iran}, title_fa = {}, abstract_fa ={}, keywords_fa = {}, doi = {10.18869/acadpub.vacres.1.1.35}, url = {http://vacres.pasteur.ac.ir/article-1-26-en.html}, eprint = {http://vacres.pasteur.ac.ir/article-1-26-en.pdf}, journal = {Vaccine Research}, issn = {2383-2819}, eissn = {2423-4923}, year = {2014} }