BACKGROUND Herpes simplex virus type 1 is one of the most common viruses among human population. ... more BACKGROUND Herpes simplex virus type 1 is one of the most common viruses among human population. Studies demonstrate the essential role of cell mediated immunity, especially CD8+ T cells, in prevention and clearance of HSV1. OBJECTIVE It is of great importance to improve our knowledge about the kinetics of CTL responses to primary and secondary HSV-1 infection. METHODS Using a sensitive technique for detection and analysis of CD8+ T cells, granzyme B ELISA, the CTL activity in the spleens of Balb/c mice at various time points after intraperitoneal administration of HSV1 (strain KOS) in primary and secondary infections were determined. RESULTS During acute HSV-1 infection, virus specific cytotoxic T cells were detected at day 5 post virus inoculation and peaked at day 7. Six hours after secondary infection the activity of memory CD8+ T cells was detected and peaked at 12 hours post infection. CONCLUSION The peak of CTL activity was found to be day 7 post infection in primary HSV-1 in...
SARS-CoV-2 seroprevalence in asymptomatic or mild symptomatic people and symptomatic patients with negative PCR results: The hidden perspective in epidemiological reports
Background: SARS-CoV-2 has led to the current pandemic of respiratory disease. The reports of con... more Background: SARS-CoV-2 has led to the current pandemic of respiratory disease. The reports of confirmed COVID-19 cases based on molecular tests do not completely cover the total number of infected people. These reports do not include the asymptomatic or mildly symptomatic patients and also the patients with false-negative RT-PCR results, while the infection is contagious in all of these conditions. Objective: In this study, we tried to improve our conception of the hidden perspective of SARS-CoV-2 in epidemiological reports. Methods: From May 30 to June 17, 2020, blood samples were collected from two groups of people: asymptomatic or mild symptomatic volunteer participants and severe symptomatic hospitalized patients with negative PCR results. Detection of SARS-CoV-2 antibody was done with ELISA kit targeting N or S proteins. Results: Totally 716 samples from volunteer participants and 81 samples from symptomatic hospitalized patients with negative PCR were evaluated. The test perfo...
INTRODUCTION Rabies is an acute viral disease that causes encephalomyelitis in mammals and human.... more INTRODUCTION Rabies is an acute viral disease that causes encephalomyelitis in mammals and human. The only way to prevent this disease is through vaccination before or after exposure. The aim of this study is to evaluate the efficiency of the Pasteur virus (PV) minigenome, using PV strain. MATERIALS AND METHODS Enhanced Green Fluorescent Protein (EGFP) sequence was placed between the designed necessary elements (Hammerhead, HDV ribozyme, 3' Leader, and 5' Trailer sequences), which resemble the rabies virus PV strain (PV2061) genome and anti-genome. These constructs were placed between T7 polymerase promoter and T7 polymerase terminator sequences. The accuracy of the minigenome was confirmed by the expression of EGFP using the helper virus in T7-BHK cell line. RESULTS The viral necessary elements of positive and negative sense strands were evaluated for the ability of EGFP expression in the presence of the helper virus. While the positive strand showed background results, no ...
Inactivation of herpes simplex virus type 1 & adenovirus type 5 by direct electric current at a biocompatible level in vitro
Clinical laboratory, 2011
BACKGROUND In vitro studies were conducted to quantify the effectiveness of low-level direct elec... more BACKGROUND In vitro studies were conducted to quantify the effectiveness of low-level direct electric current (DC) on infectivity of Herpes Simplex Virus type 1 (HSV-1), Adenovirus type 5 (AdV-5), and on viability of Vero cells. METHODS Both viruses and Vero cells were exposed to DC by using platinum electrodes connected to a DC source, then the viral infectivity and cell viability were assessed by plaque and MTT assay, respectively. RESULTS The results showed that both viruses were inactivated completely by 200 microA DC in 10 minutes (current density = 20 microA/mm2) while this amount of DC had no significant changes on the viability of Vero cells (viability > 90 %). Inactivation degree of HSV-1 and AdV-5 was 5 and 4 log per mL, respectively. Further study is required to investigate the mechanism of inactivation by this method. CONCLUSIONS DC at a biocompatible level showed the competency to inactivate the viruses in the solution completely. So it is a useful tool for designing...
Much attention is presently focused on the vaccination with certain epitopes of an antigen. To fu... more Much attention is presently focused on the vaccination with certain epitopes of an antigen. To further study the ability of neutralizing epitopes mapped in the first 1515 nucleotides of glycoprotein B of herpes simplex virus type-1 (gB-1) to induce neutralizing antibodies, a DNA immunization approach was employed. Vaccination of mice with a plasmid expressing the neutralizing epitopes induced humoral immune responses, although the antibody titre was significantly lower than that of antibodies induced by the full-length gB-1 gene. Furthermore, the plasmid DNA could not protect the mice against HSV-1 lethal challenge, but could significantly prolong the survival time compared to mockvaccinated group.
BACKGROUND Studies on efficacy of various vaccines that prevent or reduce the primary and recurre... more BACKGROUND Studies on efficacy of various vaccines that prevent or reduce the primary and recurrent HSV-1 infection have demonstrated the importance of cellular immunity for protection against the infection. We previously used DNA vaccination to induce cellular immunity against HSV-1 infection in mice. OBJECTIVE The aim of our study was to evaluate the effect of LIGHT; a member of TNF super family, on the kinetic of CTL response induced by HSV-1 glycoprotein B based DNA vaccine. METHODS Using a granzyme B ELISA for detection and analysis of CD8+ T cells, CTL activity was determined in the spleen of BALB/c mice at various time points after primary and booster dose of vaccination. The kinetics of CTL response to primary and secondary HSV-1 infection and DNA vaccination were compared to those induced by DNA vaccination in combination with LIGHT adjuvant in the present study. RESULTS In primary and secondary immunization, the CTL activity in the HSV injected group peaked 7 days and 12 h...
BACKGROUND Improving vaccine potency in the induction of a strong cell-mediated cytotoxicity can ... more BACKGROUND Improving vaccine potency in the induction of a strong cell-mediated cytotoxicity can enhance the efficacy of vaccines. Necrotic cells and the supernatant of necrotic tumor cells are attractive adjuvants, on account of their ability to recruit antigen-presenting cells to the site of antigen synthesis as well as its ability to stimulate the maturation of dendritic cells. OBJECTIVE To evaluate the utility of supernatant of necrotic tumor cells as a DNA vaccine adjuvant in a murine model. METHOD The supernatant of EL4 necrotic cells was co-administered with a DNA vaccine expressing the glycoprotein B of Herpes simplex virus-1 as an antigen model under the control of Cytomegalovirus promoter. C57BL/6 mice were vaccinated three times at two weeks intervals with glycoprotein B DNA vaccine and supernatant of necrotic EL4 cells. Five days after the last immunization, cell cytotoxicity, IFN-γ and IL-4 were evaluated. RESULTS The obtained data showed that the production of IFN-γ fr...
Background: The major problem of DNA vaccine is less immunogenicity of them verses other killed o... more Background: The major problem of DNA vaccine is less immunogenicity of them verses other killed or live whole organism vaccines therefore adjuvants for use in this kind vaccines is very necessary. Genetic adjuvants with bacterial sources are an appropriate approach to modulate immune responses to DNA vaccines. Listeria Monocytogenes proteins such as Listeriolysin O (LLO) with CD4 and CD8 epitopes can be as an adjuvant to initiate both innate and adaptive immune responses if the protein cytotoxicity can be eliminated. Herein we constructed a truncated LLO plasmid as genetic adjuvant and tested it in combination with a DNA construct as a model vaccine. Materials and Methods: About 1340bp of the 5' end of whole LLO gene was amplified by PCR on DNA purified from Listeria Monocytogenes. Sequential sub cloning of truncated LLO into the Xho I/EcoRV sites of pcDNA3.1 plasmid, downstream of CMV promoter was done. pLLO plasmid was transfected to HEK293T cell line by lipofection method. LL...
BACKGROUND Lenalidomide, a synthetic immunomodulatory drug, has a wide range of features includin... more BACKGROUND Lenalidomide, a synthetic immunomodulatory drug, has a wide range of features including anti-angiogenic and anti-proliferative properties. To date, researchers have shown that lenalidomide is capable of ameliorating the immune system factors and antitumor responses. Most researchers have reported that lenalidomide enhances the immune response in certain cancer patients through several pathways including the stimulation of Natural Killer cells; notwithstanding, it is still crucial to investigate the effect of lenalidomide on the activity of NK cell cytotoxicity both in vitro and in vivo. OBJECTIVE To evaluate the in vitro impact of lenalidomide, of different doses, on NK cytotoxicity activity and an in vivo investigation to find the adjuvant behavior of lenalidomide. METHODS NK cytotoxocity was measured with the lactate dehydrogenase (LDH) release assay via K562 cells. Lenalidomide was prepared at 1 mM, 2 mM, 4 mM and 8 mM for in vitro study. In addition, the adjuvant prop...
Background and Aims: Interferon alpha is an effective cytokine in viral infections, where it has ... more Background and Aims: Interferon alpha is an effective cytokine in viral infections, where it has various roles in immune function. The use of this antiviral agent in the treatment of viral infections and even cancers is common, although, the beneficial effects of this antiviral agent in high doses can be associated with side effects that limit its use. In this project, we tried to investigate the effects of different doses and timing of interferon alpha treatment on the expression of downstream interferon signaling genes and evaluation of the antiviral effects in patients with chronic hepatitis C. Materials and Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from treatment-naive HCV-infected patients. The cells were treated with different doses of Interferon alpha 2a (IFN-α 2a) and the mRNA expression of target genes (ISG15, MXA, PKR and OAS) at different time points was evaluated by Real Time PCR. The levels of ISG15 and OAS were measured in culture supernatant us...
Background: Autophagy suppression recently has been known to have a remarkable effect for cellula... more Background: Autophagy suppression recently has been known to have a remarkable effect for cellular adjustment and viability in the final stages of cancer. On the other hand, autophagy has the potential effect in preventing many viruses from replication. Beclin1 is the most substantial constituent in autophagy apparatus regulation. This study was intended to investigate the beclin1 siRNA knockdown effect on the extent of activity of the oncolytic vesicular stomatitis virus (VSV) as a model in cell culture. Materials and Methods: In the current study, the cancer cell line , HeLa (cervical squamous cancer cell line ) was infected by VSV, followed by beclin1 siRNA vector transfection. The potential change in the expressions of gene beclin1 in transfected cells, as well as untransfected ones were examined by real time PCR, and also the titer of viruses was compared in cells with and without transfection. Results: The results revealed that the amount of putative gene beclin1 expression in...
CD81 belongs to thetetraspanin family of cell-surface proteins, which contain four transmembrane ... more CD81 belongs to thetetraspanin family of cell-surface proteins, which contain four transmembrane domains and two outer-membrane loops. CD81 has been shown to be up-regulated in synoviocytes in rheumatoid arthritis (RA) and to promote synoviolin expression in the progression of RA. Recently, we showed that a small interfering RNA (CD81siRNA) targeting the gene encoding CD81 ameliorates arthritis in collagen-induced arthritic (CIA) rats. CD81 siRNA also decreased the expression of TNF-α in SW982 cells. Additionally, we established hybridoma cell lines producing monoclonal antibodies (MAbs) against CD81 and demonstrated that some of these MAbs impaired the proliferation of C6 glioma cells. Here, we report cloning of the cDNA encoding one such MAb (H chain and L chain) from hybridoma cells and construction of mouse-rat chimeric genes for expression of the chimeric antibody. This chimeric antibody, as expressed by CHO cells, bound to CD81 at the target's outer-membrane loop (LEL).
International Journal of Molecular and Cellular Medicine (IJMCM), 2020
A growing area of research is focused on cancer therapy, and new therapeutic approaches are welco... more A growing area of research is focused on cancer therapy, and new therapeutic approaches are welcomed. Mesenchymal stem cell (MSC)-based gene therapy is a promising strategy in oncology. Intrinsic tropism and migration to tumor microenvironment with off lights are attractive features of this type of cell carrier. In this way, suicide genes have also found a good platform for better performance and have shown a stronger anti-tumor mechanism by riding on mesenchymal cells. In this study, we investigated the anti-tumor activity of intratumoral injected MSCs transduced with a lentivector expressing the HSV/TK in a mouse cervical cancer model. Following the injection of MSCs transduced with lentivector carrying TK, MSCs alone or PBS into the mice tumor, ganciclovir was administered intraperitoneally during 14 days, and tumor size, survival time, natural killer (NK) cells and cytotoxic T lymphocyte (CTL) activities were assessed. We demonstrated that combination of suicide therapy and cell...
Nucleic acid immunization has recently exhibited a great promise for immunotherapy of various dis... more Nucleic acid immunization has recently exhibited a great promise for immunotherapy of various diseases. However, it is now clear that powerful strategies are imminently needed to improve their efficiency. In this regard, whole bacteriophage particles have been described as efficient DNA vaccine delivery vehicles, capable of circumventing the limitations of naked DNA immunization. Moreover, phage particles could be engineered to display specific peptides on their surfaces. Given these inherent characteristics of phages, we have designed a novel hybrid phage-DNA immunization vector using both M13 and pAAV plasmid elements. Following the construction and in vitro confirmation of the designed vectors, they were used for comparative mice immunization, carrying the same DNA sequence. The results indicated the efficacy of the designed hybrid phage particles, to elicit higher humoral immunity, in comparison to conventional DNA-immunization vectors (pCI). In light of these findings, it could be concluded that using adeno-associated virus (AAV) expression cassette along with displaying TAT peptide on the surface of the phage particle could be deemed as an appealing strategy to enhance the DNAimmunization and vaccination efficacy.
Although several years have passed since the determination of the human papilloma virus (HPV) as ... more Although several years have passed since the determination of the human papilloma virus (HPV) as the causative agent for cervical cancer, a definitive treatment has not yet been found. Interferon alpha (IFN α) immunotherapy is one of the promising methods for tumor treatment, although numerous side effects were observed in clinical trials. Recently, a new type of interferon, lambda interferon (IFN λ), has been discovered with fewer side effects than IFN α since its receptor repertoire is limited. IFN λ has a series of activities including antiviral, anti proliferative and anti tumor actions. In the present study, the effects of IFN α and IFN λ on the TC1 papilloma tumor model in C57BL/6 mice were evaluated. TC1 cells were injected into the mice subcutaneously. Upon tumor formation, murine IFN, mIFN α and mIFN λ, expres sion plasmids were injected intratumorally in combination or alone. The survival time and tumor size as well as apoptosis in tumors and NK cytoxicity were measured after three injections. As compared with the control group, the remarkable results especially in the group which received mIFN α and mIFN λ together were obtained for all of the measured parameters. Although IFN λ is a new member of the interferon family and its properties should be studied in detail, the data obtained suggests that the use of IFN λ especially in combi nation with IFN α could be considered as an effective strategy for papilloma cervical cancer immunotherapy.
Background and Aims: The aim of this study was cloning and expression of rabies virus glycoprotei... more Background and Aims: The aim of this study was cloning and expression of rabies virus glycoprotein by a eukaryotic expression plasmid pcDNA3.1(+) in BSR cell line. This construct might be used for a potential DNA vaccine. Materials and Methods: Glycoprotein gene was synthesized and cloned into pBluescript vector and then sub cloned into eukaryotic expression vector (pcDNA3.1(+)). After verification of the cloning, the recombinant plasmid was transfected into BSR cell line (a clone of BHK-21 cell), and its expression was detected by RT-PCR. Results: The authenticity of the recombinant plasmid pcDNA3.1(+)-Gp has been confirmed by a quick check method and restriction endonuclease digestion analysis, and after transfection into eukaryotic cells, the presence of mRNA transcripts was verified by reverse transcriptase-polymerase chain reaction (RT-PCR). Conclusion: This study demonstrated that the construction of eukaryotic expression plasmid for rabies virus glycoprotein is possible. Nevertheless, more work is necessary to develop this kind of vaccine for final use.
Asian Pacific Journal of Allergy and Immunology, 2015
Background and Objectives: Approximately onethird of hepatitis C virus (HCV) infected patients wh... more Background and Objectives: Approximately onethird of hepatitis C virus (HCV) infected patients who complete antiviral therapy with undetectable serum HCV RNA at the end of therapy (ETR), will experience relapse. The reasons for the failure of treatment have not been elucidated. It was showed that HCV RNA can persist and replicate in extra hepatic sites, e.g. in peripheral blood mononuclear cells (PBMCs), but the relevance of its presence with relapse over time is still unknown. Moreover, interferon-gamma (IFN-γ) and IFN-lambdas [IFN-λ1, interleukin-29 (IL-29)], possess potent antiviral activity. We studied if the presence of plus-/minus strand RNA in PBMCs of patients and the serum level of IFN-γ and IL-29, which is the most abundant IFN-lambdas in serum, can be considered as predictive factors in relapse outcomes. Methods: Patients were screened for plus-/minus strand RNA at ETR and after 6 months. Also, we measured the serum level of IFN-γ and IL-29 and compared the result with those who developed a sustained virological response (SVR). Results: Levels of IL-29 and IFN-γ serum were significantly higher in SVR at ETR and 6 months later compared to those of the relapsed patients, but there was no difference between the two groups regarding the presence or absence of plus-/minus HCV strand in PBMCs. Conclusions: Our novel findings showed that the serum level of IL-29 and IFN-γ are predictive of relapse outcomes to HCV treatment, but there was no association between the presence of plus-/minus HCV RNA in PBMCs of patients with an outcome of therapy at ETR and later.
Background: Taqman one-step real-time PCR (RT-PCR) has special importance due to its high sensiti... more Background: Taqman one-step real-time PCR (RT-PCR) has special importance due to its high sensitivity and specificity in the diagnosis of infectious diseases such as viral infections. In the recent pandemic of SARS-CoV-2, diagnostic kits based on this method are commonly used for molecular detection. One of the main systematic errors that misinterpret the results is using inaccurate internal control in RT-PCR diagnostic kits. Designing primers and probes that span exon-exon junction will avoid genomic DNA amplification and lead to obtaining high specific results. Objectives: This study aimed to evaluate the endogenous internal control of primers and probe for RNase P RNA to reduce false-negative results in respiratory samples. Methods: In this study, 30 samples of patients who were negative for SARS-CoV-2, influenza A, and influenza B were re-evaluated for SARS-CoV-2 using newly designed primers and probes for RNase P RNA (ultra-specific primers and probe). We also performed bioinfo...
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Papers by Taravat Bamdad