Azam Bolhassani

Pasteur Institute of Iran (IPI), Teheran, Tehrān, Iran

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Publications (74)110.89 Total impact

  • Kimia Kardani · Azam Bolhassani · Sepideh Shahbazi
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    ABSTRACT: The essential goal of vaccination is to generate potent and long-term protection against diseases. Among different vaccine modalities, prime-boost vaccine strategies could enhance cellular and also humoral immunity in several animal models. These strategies have been applied for the development of vaccines against important infectious diseases such as HIV, SIV, HCV, HSV, and HBV indicating promising results even in clinical trials. Several factors including selection of antigen, type of vector, delivery route, dose, adjuvant, boosting regimen, the order of vector injection, and the intervals between different vaccinations influence the outcome of prime-boost immunization approaches. The reported data suggest that the prime-boost strategy as a combination of vaccines (i.e., heterologous prime-boost) may be better than a single vaccine for protection against infectious diseases. Indeed, in many cases, heterologous prime-boost can be more immunogenic than homologous prime-boost strategy. This review discusses the recent advances in prime-boost immunization strategies as well as their benefits and mechanisms of action.
    No preview · Article · Dec 2015 · Vaccine
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    ABSTRACT: One of the significant problems in vaccination projects is the lack of an effective vaccine against hepatitis C virus (HCV). The goal of the current study is to evaluate and compare two DNA constructs encoding HCV core and coreE1E2 genes alone or complexed with MPG peptide as a delivery system for stimulation of antibody responses and IFN-γ secretion in Balb/c mice model. Indeed, MPG cell penetrating peptide was used to improve DNA immunization in mice. Our results demonstrated that MPG forms stable non-covalent nanoparticles with pcDNA-core and pcDNA-coreE1E2 at an N/P ratio of 10:1. The in vitro transfection efficiency of core or coreE1E2 DNA using MPG and TurboFect delivery systems was confirmed by western blot analysis. The results indicated the expression of the full-length core (∼21 kDa), and coreE1E2 (∼83 kDa) proteins using an anti-His monoclonal antibody. In addition, the expression of HCV core and coreE1E2 proteins was performed in bacteria and the purified recombinant proteins were injected to mice with Montanide 720 adjuvant. Our data showed that the immunized mice with HCV core and coreE1E2 proteins generated the mixture of sera IgG1 and IgG2a isotypes considerably higher than other groups. Furthermore, DNA constructs encoding core and coreE1E2 complexed with MPG could significantly induce IFN-γ secretion in lower concentrations than the naked core and coreE1E2 DNAs. Taken together, the DNA formulations as well as protein regimens used in this study triggered high-level IFN-γ production in mice, an important feature for the development of Th1 immune responses.
    No preview · Article · Nov 2015 · Drug Delivery
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    ABSTRACT: Development of an effective vaccine against HIV-1 infection is a main concern in worldwide. A potent vaccine for HIV-1 requires the induction and maintenance of both humoral and cellular immunity. In this study, the levels of humoral and cellular immune responses were compared using MPER-V3 injection in three immunization strategies such as DNA/DNA, peptide/peptide, and DNA/ peptide (prime-boost). MPG peptide and Montanide 720 were used as a DNA delivery system, and as a peptide adjuvant, respectively. Our results demonstrated that MPG forms stable non-covalent nanoparticles with plasmid DNA at N/P ratio of 10:1 (∼ 110-130nm). The in vitro transfection efficiency of MPER-V3 DNA using MPG was comparable with lipofectamine and turbofect reagents as a common delivery system. In vivo prime-boost immunization using HIV-1 MPER-V3 could significantly enhance humoral and cellular immune responses as compared to control groups. The mixture of IgG1 and IgG2a was observed for each strategy, but IFN-γ production was significantly higher in prime-boost and peptide immunizations than that in DNA immunizations, inducing Th1 response. Moreover, our data showed that prime immunization with low dose of the nanoparticles (MPER-V3 DNA: MPG at ratio of 1:10) followed by MPER-V3 peptide drives T cell responses towards a Th1-type similar to high dose of the naked DNA prime/ peptide boost immunization. Generally, the prime-boost strategy could improve both immune responses against MPER and especially V3 peptides suggesting its application as a promising HIV vaccine candidate in future.
    No preview · Article · Oct 2015 · Immunology letters
  • Zeinab Shirbaghaee · Azam Bolhassani
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    ABSTRACT: Virus-like particles (VLPs) mimic the whole construct of virus particles devoid of viral genome as used in subunit vaccine design. VLPs can elicit efficient protective immunity as direct immunogens compared to soluble antigens co-administered with adjuvants in several booster injections. Up to now, several prokaryotic and eukaryotic systems such as insect, yeast, plant, and E. coli were used to express recombinant proteins, especially for VLP production. Recent studies are also generating VLPs in plants using different transient expression vectors for edible vaccines. VLPs and viral particles have been applied for different functions such as gene therapy, vaccination, nanotechnology, and diagnostics. Herein, we describe VLP production in different systems as well as its applications in biology and medicine. This article is protected by copyright. All rights reserved.
    No preview · Article · Oct 2015 · Biopolymers
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    ABSTRACT: Background: The association between human papillomavirus (HPV) infections and cervical cancer has suggested the design of prophylactic and therapeutic vaccines against genital warts. The HPV capsid has made of two L1 and L2 coat proteins that have produced late in viral infections. Regarding to the recent studies, two commercial prophylactic vaccines have based on L1 viral like particles (VLPs) could strongly induce antibody responses, and protect human body from HPV infections. However, the use of these HPV vaccines has hindered due to their high cost and some limitations. Currently, among various vaccination strategies, live vector-based vaccines have attracted a great attention. Objectives: Herein, a non-pathogenic strain of the protozoan organism known as Leishmania tarentolae has utilized to induce potent humoral immunity in mice model. Materials and methods: At first, cloning of HPV16 L1 gene into Leishmania expression vector has performed and confirmed by PCR and digestion with restriction enzymes. The promastigotes of Leishmania tarentolae (L.tar) have transfected with linearized DNA construct by electroporation. Protein expression has analyzed by SDS-PAGE and western blotting. Then, the immunogenicity of leishmania expressing L1 protein (L.tar-L1) has assessed in mice model. Results: Our data has indicated that subcutaneous immunization of mice with the recombinant L.tar-L1 has led to enhance the levels of IgG1 and lgG2a in comparison with control groups. Furthermore, there was no significant increase in antibody levels between two and three times of immunizations. Conclusions: The recombinant live vector was able to induce humoral immunity in mice without need of any adjuvant. However, further studies have required to increase its efficiency.
    Preview · Article · Oct 2015
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    Nasiri V · Dalimi A.H · Ghaffarifar F · Bolhassani A
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    ABSTRACT: Vaccination efficacy of prime-boost strategy based on live recombinant Leishmania tarentolae expressing kmp-11 – ntgp96-gfp fusion against visceral leishmaniasis Vahid Nasiri 1, Abdolhossein Dalimi 2, Fatemeh Ghaffarifar 2, Azam Bolhassani 3 1Department of Parasitology, Razi Vaccine and Serum Research Institute, Alborz, Iran 2Department of Parasitology, Faculty of Medicine, Tarbiat Modares University, Tehran, Iran 3Department of Hepatitis & AIDS, Pasteur Institute of Iran, Tehran, Iran Introduction: Leishmaniases are neglected tropical diseases that cause human infections varying from self-healing cutaneous lesions to mucosal diffuse cutaneous and visceral forms. Kinetoplastid membrane protein-11 is a fully protective Leishmania antigen and NT-Gp96 acts as a strong biologic immunologic adjuvant. The use of the Leishmania tarentolae as a live vaccine vector to deliver specific Leishmania antigens is a recent approach and we used this species as a live vaccine vector to deliver these specific genes fusion using Live/Live prime-boost strategy in protecting BALB/c mice against visceral Leishmaniasis. Methods: L. infantum KMP-11 and NT-Gp96 cloned into the pJET1.2/blunt vector and then the KMP-11, NT-Gp96 and GFP fused in pEGFP-N1 and subcloned into Leishmanian pLEXSY-neo vector. Finally, this construct transferred to Leishmania tarentolae by electroporation. Tranfection confirmed by SDS-page, WESTERN blot, flowcytometry and RT-PCR. Protective Efficacy of recombinant Leishmania tarentolae was tested with single and prime-boost vaccination strategy in susceptible BALB/c mice. 3 weeks after the last immunization, the mice were challenged with stationary phase promastigote of L. infantum by intraperitoneal injection. Both humoral and cellular immune responses were assessed before and at 4 and 10 weeks after Leishmania challenge. Results: KMP- NT-Gp96-GFP Fusion cloned successfully into pLEXSY-neo vector and this construct successfully transferred to Leishmania tarentolae. The strongest protective effect was observed with L. tarentolae- KMP- NT-Gp96-GFP by prime-boost strategy. Discussion: The present study is the first to use a combination of leishmania antigen with a human immunologic antigen in live recombinant L. tarentolae with this vaccination strategy that could provide a potent rout for future vaccine development. Keywords: KMP-11, NT-GP96, Leishmania infantum, vaccine.
    Full-text · Conference Paper · Aug 2015
  • Azam Bolhassani · Zeinab Shirbaghaee · Elnaz Agi · Noushin Davoudi
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    ABSTRACT: Viral like particles (VLPs) have been used as immunogen for improvement of preventive vaccines against several viral infections in preclinical and clinical trials. These constructs can stimulate both cellular and humoral immunity. Two prophylactic HPV L1 VLP vaccines known as Gardasil and Cervarix were commercialized worldwide. However, there are main problems for expression and purification of VLPs in eukaryotic expression systems such as baculovirus and yeast leading to high cost of these vaccines. A novel Leishmania protozoan system has been applied to produce different recombinant proteins due to unique properties including generation of similar proteins with mammalian, easy handling, and large-scale culture. In the current study, we developed a novel strategy to produce HPV L1 VLP using stably transfected Leishmania cells. The positive transfectants were analyzed by SDS-PAGE and Western blot analysis. The assembly of purified L1 protein was detected by TEM microscopy. Finally, C57BL/6 mice were immunized by crude VLPs and antibody responses were assessed. The results of electronic microscopy revealed average 55-60 nm for L1 VLP. Furthermore, high IgG1 and IgG2a antibody responses were generated by L1 VLPs in mice similar to L1 VLPs produced in baculovirus-infected insect cells. Regarding the results, the amount of recombinant protein generated by leishmania was 2-3 mg/ 500 ml media, suggesting further optimization of this system for using in large animals and human. Copyright © 2015. Published by Elsevier Inc.
    No preview · Article · Aug 2015 · Protein Expression and Purification
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    Kimia Kardani · Golnaz Mardani · Azam Bolhassani

    Preview · Article · Aug 2015
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    ABSTRACT: DNA vaccines against human papillomavirus (HPV) type 16 have not been successful in clinical trials, due to the lack of an appropriate delivery system. In this study, a peptide-based gene delivery system, MPG, which forms stable non-covalent nanoparticles with nucleic acids, was used for in vitro and in vivo delivery of HPV16 E7 DNA as a model antigen. The results demonstrated that at Nitrogen/Phosphate (N/P) ratio over 10:1, this peptide can effectively condense plasmid DNA into stable nanoparticles with an average size of 180-210nm and a positive surface charge. The transfection efficiency of MPG-based nanoparticles was shown to be comparable with Polyethyleneimine (PEI). The efficient protein expression detected by western blotting and flow cytometry supports the potential of MPG-based nanoparticles as a potent delivery system in DNA vaccine formulations. Immunization with MPG/E7DNA nanoparticles at an N/P ratio of 10:1 induced a stronger Th1 cellular immune response with a predominant interferon-γ (IFN-γ) profile than those induced by E7DNA alone in a murine tumor model. These findings suggest that MPG peptide as a novel gene delivery system could have promising applications in improving HPV therapeutic vaccines. Copyright © 2015. Published by Elsevier Ltd.
    Full-text · Article · May 2015 · Vaccine

  • No preview · Article · May 2015 · Human Vaccines and Immunotherapeutics
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    ABSTRACT: Delivery of the macromolecules including DNA, miRNA, and antisense oligonucleotides is typically mediated by carriers due to the large size and negative charge. Different physical (e.g., gene gun or electroporation), and chemical (e.g., cationic polymer or lipid) vectors have been already used to improve the efficiency of gene transfer. Polymer-based DNA delivery systems have attracted special interest, in particular via intravenous injection with many intra- and extracellular barriers. The recent progress has shown that stimuli-responsive polymers entitled as multi-functional nucleic acid vehicles can act to target specific cells. These non-viral carriers are classified by the type of stimulus including reduction potential, pH, and temperature. Generally, the physicochemical characterization of DNA-polymer complexes is critical to enhance the transfection potency via protection of DNA from nuclease digestion, endosomal escape, and nuclear localization. The successful clinical applications will depend on an exact insight of barriers in gene delivery and development of carriers overcoming these barriers. Consequently, improvement of novel cationic polymers with low toxicity and effective for biomedical use has attracted a great attention in gene therapy. This article summarizes the main physicochemical and biological properties of polyplexes describing their gene transfection behavior, in vitro and in vivo. In this line, the relative efficiencies of various cationic polymers are compared. This article is protected by copyright. All rights reserved. © 2015 Wiley Periodicals, Inc.
    No preview · Article · Mar 2015 · Biopolymers
  • Sahar Hosseinzadeh · Azam Bolhassani
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    ABSTRACT: One approach to improve the vaccine quality is the incorporation of immunomodulators and/ or adjuvants with modified delivery systems. The use of delivery systems especially chemical carriers is a promising strategy in the prevention and treatment of infections, cancers, allergies and autoimmune diseases. These systems are able to elicit an effective immune response as well as stability and safety in vaccine development. Synthetic microparticles, liposomes, chitosan, virus like particle, polymeric nanogel, phytosome, noisome, and micro/ nanospheres have been applied as carriers, providing a broad variety of immunomodulatory effects in vaccines. The potency and nature of immune responses rely on the physicochemical properties of the vaccine constructs (e.g., size and charge), the route of injection, the biochemical characteristics and the amount of antigen. Three main steps are necessary for vaccine efficiency such as targeting, activation and transfection/ antigen presentation. These systems can generally influence the type and direction of immune responses. This review describes different vaccine delivery systems developed to generate immunomodulatory effects.
    No preview · Article · Mar 2015 · Current Drug Delivery
  • Azam Bolhassani
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    ABSTRACT: The use of specific compounds to suppress the growth of tumors or reverse carcinogenesis is defined as chemoprevention. Natural products have been known as one of the most important resources of anticancer agents. Among them, carotenoids are lipophilic molecules accumulating in lipophilic compartments including lipoproteins and/or membranes. Various carotenoids were used as major phytonutrients to inhibit the development of tumors in vitro and in vivo. They have shown different functions such as scavenging free radicals, inhibition of angiogenesis, prevention of cell propagation, and apoptosis induction in lung, colon, breast and prostate. Regarding to these roles, most carotenoids possess anti-oxi¬dant properties. However, their therapeutic use is problematic due to the lack of solubility of carotenoids in water. Hence, the recent studies have been focused on uncommon carotenoids soluble in water because of their glycosylated form, such as crocin(s) extracted from saffron. These structures with their cytotoxicity effects on human cancer cells are suggested as the most suitable compounds for cancer treatment. Herein, we summarize different functions of carotenoids for suppressing tumor growth.
    No preview · Article · Mar 2015 · Anti-Cancer Agents in Medicinal Chemistry (Formerly Current Medicinal Chemistry - Anti-Cancer Agents)
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    ABSTRACT: Backgrounds: It has been known that Kinetoplastid membrane protein-11 is a fully protective antigen and also NT-Gp96 acts as a strong biologic immunologic adjuvant. We used non-pathogenic Leishmania tarentolae as a live vaccine vector to deliver these specific genes against cutaneous leishmaniasis. Materials and Methods: L. major KMP-11 and NT-Gp96 cloned into the pJET1.2/blunt and pEGFP-N1 vectors and then the KMP-11, NT-Gp96 and GFP fused in pEGFP-N1 and subcloned into Leishmanian pLEXSY-neo vector. Finally, this construct transferred to Leishmania tarentolae by electroporation. Tranfection confirmed by SDS-page, WESTERN blot, flowcytometry and RT-PCR. Protective Efficacy of recombinant Leishmania tarentolae Combined with Naloxone were tested h in susceptible BALB/c mice. Both humoral and cellular immune responses were assessed before challenge and at 4 weeks after Leishmania infection. The results: KMP- NT-Gp96-GFP Fusion cloned successfully into pLEXSY -neo vector and this construct successfully transferred to Leishmania tarentolae. The strongest protective effect was observed with live recombinant L. tarentolae with Naloxone. Conclusions: The present study is the first to use a combination of live recombinant L. tarentolae with Naloxone and this vaccination strategy could provide a potent rout for future vaccine development.
    No preview · Conference Paper · Feb 2015
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    Full-text · Article · Feb 2015 · Iranian Journal of Biotechnology
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    ABSTRACT: Backgrounds: : Adjuvants are immunological agents that added to vaccine to modify the immune response .It has been known that KMP-11 is a fully protective antigen and NT-Gp96 acts as a strong biologic immunologic adjuvant. We used non-pathogenic Leishmania tarentolae to deliver these specific genes and combination of it with Alum was evaluated against cutaneous Leishmaniasis. Materials and Methods: Efficacy of recombinant Leishmania tarentolae and combination of it with Alum against cutaneous Leishmaniasis was tested in susceptible BALB/c mice. Both humoral and cellular immune responses were assessed before and at 4 weeks after challenge with Leishmania major infection. The results: The strongest protective immunity were observed with live recombinant L. tarentolae alone but it seems that Alum combination could not protect against infection. Conclusions: This study is the first to use a combination of live non pathogenic recombinant L. tarentolae parasite with Alum adjuvants and seems that this vaccination strategy could not provide a potent rout for future vaccine development.
    Full-text · Conference Paper · Jan 2015
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    ABSTRACT: Backgrounds: Adjuvants are used to give a higher and longer lasting protection immunity response .It has been known that KMP-11 is a fully protective antigen and NT-Gp96 acts as a strong biologic immunologic adjuvant. We used non-pathogenic Leishmania tarentolae to deliver these specific genes and combination of it with Liquid Paraffin was evaluated against cutaneous Leishmaniasis. Materials and Methods: Efficacy of recombinant Leishmania tarentolae and combination of it with Liquid Paraffin was tested against cutaneous Leishmaniasis in BALB/c mice. Both humoral and cellular immune responses were assessed before and at 4 weeks after challenge with Leishmania major infection. The results: The strongest protective immunity against infection was observed with live recombinant L. tarentolae and combination of it with Liquid Paraffin. Conclusions: This is the first to use a combination of live non pathogenic recombinant L. tarentolae parasite with Liquid Paraffin adjuvants and seems that the use of these adjuvants with live cells can lead to better and longer contact of vaccine agents to the immune system and then result to better stimulation of immunity and provide a potent rout for future vaccine development.
    No preview · Conference Paper · Jan 2015
  • Nasiri V · Dalimi A.H · Ghaffarifar F · Bolhassani A · Karimi Gh

    No preview · Conference Paper · Jan 2015
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    ABSTRACT: The development of an efficient vaccine against high-risk HPV types can reduce the incidence rates of cervical cancer by generating anti-tumor protective responses. Traditionally, the majority of prophylactic viral vaccines are composed of live, attenuated or inactivated viruses. Among them, the design of an effective and low-cost vaccine is critical. Inactivated vaccines especially heat-killed yeast cells have emerged as a promising approach for generating antigen-specific immunotherapy. Recent studies have indicated that yeast cell wall components possess adjuvant activities. Moreover, a non-pathogenic protozoan, Leishmania tarentolae (L.tar) has attracted a great attention as a live candidate vaccine. In current study, immunological and protective efficacy of whole recombinant killed Pichia pastoris and Leishmania tarentolae expressing HPV16 L1 capsid protein was evaluated in tumor mice model. We found that Pichia-L1, L.tar-L1 and Gardasil groups increase the IgG2a/IgG1 ratio, indicating a relative preference for the induction of Th1 immune responses. Furthermore, subcutaneous injection of killed Pichia-L1 generated the significant L1-specific IFN-γ immune response as well as the best protective effects in vaccinated mice as compared to killed L.tar-L1, killed Pichia pastoris, killed L.tar and PBS groups. Indeed, whole recombinant Leishmania tarentolae could not protect mice against C3 tumor mice model. These data suggest that Pichia-L1 may be a candidate for the control of HPV infections.
    No preview · Article · Dec 2014 · Human Vaccines and Immunotherapeutics
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    ABSTRACT: Saffron and its components have been suggested as promising candidates for cancer prevention. Carotenoids and monoterpene aldehydes are two potent ingredients of saffron. The goal of the current study was to investigate the anti-tumor effect of chemo-immunotherapy using saffron and its ingredients followed by E7-NT (gp96) DNA vaccine against tumors expressing the E7 protein of human papillomavirus. The in vitro cytotoxic and apoptotic effects of aqueous saffron extract and its components were evaluated in malignant TC-1 and non-malignant COS-7 cell lines. Then, multimodality treatments using E7-NT (gp96) DNA vaccine combined with saffron extract and its ingredients as well as single-modality treatments were tested for their efficacy in inhibiting large and bulky tumor growth. Saffron and its components exerted a considerable anti-tumor effect through prevention of cell growth and stimulation of programmed cell death. Furthermore, 100 % of mice treated with crocin were tumor-free, in contrast to DNA vaccine alone (~66.7 %) and DNA + crocin (~33.3 %) indicating the high potency of crocin as a chemotherapeutic agent. Interestingly, the multimodality treatment using DNA vaccine along with picrocrocin augmented the anti-tumor effects of picrocrocin. Thus, the combination of DNA vaccine with saffron extract and crocin at certain concentrations did not potentiate protective and therapeutic effects compared to mono-therapies for the control of TC-1 tumors.
    No preview · Article · Nov 2014 · Archives of Virology