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Papers by Hoorieh Soleimanjahi

Research paper thumbnail of Humoral immunogenicity assessment after receiving three types of SARS-CoV-2 vaccine

Scientific Reports

Several vaccines have been developed against SARS-CoV-2 and subsequently approved by national/int... more Several vaccines have been developed against SARS-CoV-2 and subsequently approved by national/international regulators. Detecting specific antibodies after vaccination enables us to evaluate the vaccine’s effectiveness. We conducted a prospective longitudinal study among members of Tarbiat Modares University of Tehran, Iran, from 4 September 2021 until 29 December 2021. We aimed to compare the humoral immunogenicity of 3 vaccine types. Participants consisted of 462 adults. Anti-SARS-CoV-2 receptor-binding domain [RBD] IgG titer was compared in 3 groups, each vaccinated by available vaccines in Iran at the time: Oxford/AstraZeneca, COVIran Barekat, and Sinopharm. The median IgG titer was: 91.2, 105.6, 224.0 BAU/ml for Sinopharm, COVIran Barekat and Oxford/AstraZeneca respectively after the first dose; 195.2, 192.0, 337.6 BAU/ml after the second one. We also analyzed the frequency of antibody presence in each vaccine group, in the same order the results were 59.0%, 62.6% and 89.4% aft...

Research paper thumbnail of Prevalence of herpes simplex viruses types 1 and 2 infections among suspected children of encephalitis in Kermanshah, Iran

Iranian Journal of Microbiology

Background and Objectives: Various infectious and non-infectious factors can cause encephalitis i... more Background and Objectives: Various infectious and non-infectious factors can cause encephalitis in the central nervous system (CNS), the most important of which are viruses. Herpes viruses are one of the most important causes of encephalitis worldwide. PCR detected the virus on the cerebrospinal fluid (CSF) sample. The aim of this study was to set up an in-house PCR to identify herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) and determine the prevalence of these viruses in suspected children of encephalitis. Materials and Methods: This cross-sectional study was conducted on 160 suspected children with encephalitis cases re- ferred to Dr. Kermanshahi Children Hospital, Kermanshah, Iran, from April to March 2021. CSF samples were extracted using a viral extraction kit, and a PCR was performed. The level of glucose and total protein of the samples was measured. Results: The total prevalence of HSV was 16.25%. 17 samples were positive for HSV-1 (10.6%), and 9...

Research paper thumbnail of The Effect of Oncolytic Reovirus Infection on Nitric Oxide Secretion and Induction of Apoptosis in Adipose Tissue-Derived Mesenchymal Stem Cells

Iranian Journal of Medical Microbiology

Background and Aims: Oncolytic viruses (OVs) are a new approach in treatment of cancer. Antitumor... more Background and Aims: Oncolytic viruses (OVs) are a new approach in treatment of cancer. Antitumor efficacy of OVs were limited due to insufficient and non-specific viral delivery to tumor sites. To overcome this issue, mesenchymal stem cells (MSCs) were used for their ability to specifically homing into tumors. The main aim of this study was to use MSCs as carriers and investigate the effect of oncolytic reovirus infection in MSCs, induction of apoptosis, nitric oxide (NO) secretion and their effects for selectively killing tumor cells, to use in future. Materials and Methods: MSCs isolated from mice adipose tissue and confirmed. Then, the ability of the virus to infect MSCs and the effect of reovirus infection in induction of apoptosis and NO secretion in MSCs were evaluated. Results: The results demonstrate that reovirus could replicate on MSCs. The finding indicated that the NO production significantly was higher at 72 h post infection with different MOI in comparison to the control cells. Also, reovirus induced high level of apoptosis in the MSCs at 48 h post infection compared with the control cells. Conclusions: Based on observed results, reovirus increased the secretion of iNOS (inducible nitric oxide) in the infected MSCs at 48 h post infection; therefore, high amounts of NO and reovirus replication were found to trigger apoptosis at 48 h post infection. Therefore, by optimizing the replication time of virus in the MSCs, specific viral delivery to tumor sites are available and causes cancer cells' death.

Research paper thumbnail of The Convenient and Economical Method to Collect Adipose Mesenchymal Stem Cells

Medical laboratory sciences, 2019

Background: Enzymatic digestion is an essential stage for culturing Mesenchymal Stem Cells (MSCs)... more Background: Enzymatic digestion is an essential stage for culturing Mesenchymal Stem Cells (MSCs) and their therapeutic application. Several factors such as being cost-benefit, efficiency, safety, yield and amount of produced cells are determinant for choosing the appropriate enzyme. Collagenase is a conventional enzyme commonly used for enzymatic digestion. However, other enzymes like trypsin and even combination of these enzymes can be used as an alternative strategy in different situations. Materials and Methods: Abdominal subcutaneous adipose tissue was obtained from male BALB/c mice and digested under three different enzymatic processes: collagenase and collagenase/trypsin and trypsin. Cell culture process was performed under standard condition and MSCs at 3rd passage were used for further characterization by flow cytometry. Results: In this study, two different enzymatic methods for digestion of adipose-derived MSCs (ADSCs) of BALB/c mice were investigated. The morphology of cells was pretty different and was more homogenous in collagenase group. Also the yield of cells was varied among groups. Furthermore, the obtained data from flow cytometry revealed that ADSCs were positive for CD90 (70%), CD29 (98%), CD105 (52%) and negative for CD45 (<2%).Conclusion: Application of different enzymes depends on various conditions. Altogether, these data indicate that although use of trypsin in isolation protocol is cost-benefit, it can be used as an alternative method whenever limited number of cells will be needed. However, collagenase as a well-known and conventional method can be used for isolation of larger quantity of cells with several applications.

Research paper thumbnail of The Effect of Autophagy Induction in Oncolytic Reovirus Replication in Mesenchymal Stem Cells

International Journal of Medical Laboratory, 2021

Background and Aims: Oncolytic reoviruses can infect and kill malignant cells while sparing their... more Background and Aims: Oncolytic reoviruses can infect and kill malignant cells while sparing their normal counterparts. Reoviral infection can induce or activate autophagy, even though metformin can induce autophagy. Identifying and regulating the cellular pathways important for reovirus replication and oncolysis can improve targeted-biological therapies for cancer. Here, the autophagic process was triggered via metformin, and we investigated the effect of autophagy activation on oncolytic reovirus replication in mesenchymal stem cells as primary cells and L929 cell lines. Materials and Methods: Adipose derived mesenchymal stem cells (AD-MSCs) and L929 cells were treated with metformin and reovirus type-3 strain Dearing (T3D). Twenty-four hours after infection, the viability of AD-MSCs and L929 cells were examined by MTT assay. Also, the effect of metformin-induced autophagy in the reovirus replication in these cells was determined by real-time polymerase-chain-reaction. Results: Our...

Research paper thumbnail of Development of a Western Blot Assay for Detection of Antibodies against HSV Using Purified HSV Virions Prepared by Sucrose Density Gradient

Objective(s) Herpes simplex viruses (HSVs) have widespread and ubiquitous prevalence in the human... more Objective(s) Herpes simplex viruses (HSVs) have widespread and ubiquitous prevalence in the human population and they have received a great deal of attention due to the range of diseases, they caused as a result of an infection. It seems that the fast and reliable diagnostic methods are needed for detecting the herpes simplex virus type 1 (HSV1) antibodies especially in patients with HSV encephalitis, immunocompromised people, and neonatal infections. The aim of this study was designing a Western blotting method for HSV1 antibody detection, using the purified virus by sucrose density gradient centrifugation procedure. Materials and Methods The most reliable method for HSV detection is virus neutralization test but it needs cell culture preparation, high expertise, as well as the high amounts of serum samples. Considering the difficulties of this method, we tried to run a new one for HSV antibody detection by propagating the viruses and then purify them by sucrose density gradient centrifugation method. The purified viruses used as antigens in Western blotting assay. Results Diluted sera (1:100, and 1:200 dilutions) used in Western blotting and twofold dilutions of the sera applied in virus neutralization test. Five of twenty seven samples were negative in Western blotting and the same results obtained in virus neutralization test. Comparing with our gold standard, the sensitivity and specificity of the developed assay were both 100%. Conclusion Our results show that the designed method is a reliable method for replacing the virus neutralization test in diagnostic laboratories. It can also, be used for confirming the ELISA results.

Research paper thumbnail of Kinetics of Oncolytic Reovirus T3D Replication and Growth Pattern in Mesenchymal Stem Cells

Cell Journal (Yakhteh), 2020

Objective Currently, application of oncolytic-virus in cancer treatment of clinical trials are gr... more Objective Currently, application of oncolytic-virus in cancer treatment of clinical trials are growing. Oncolytic-reovirus is an attractive anti-cancer therapeutic agent for clinical testing. Many studies used mesenchymal stem cells (MSCs) as a carrier cell to enhance the delivery and quality of treatment with oncolytic-virotherapy. But, biosynthetic capacity and behavior of cells in response to viral infections are different. The infecting process of reoviruses takes from two-hours to one-week, depends on host cell and the duration of different stages of virus replication cycle. The latter includes the binding of virus particle, entry, uncoating, assembly and release of progeny-viruses. We evaluated the timing and infection cycle of reovirus type-3 strain Dearing (T3D), using one-step replication experiment by molecular and conventional methods in MSCs and L929 cell as control. Materials and Methods In this experimental study, L929 and adipose-derived MSCs were infected with differ...

Research paper thumbnail of Development of Poly ( A )-Tailed Universal Reverse Transcription Rearranged Rotavirus

rotaviruses are necessary for understanding the molecular and antigenic features of the virus. Ba... more rotaviruses are necessary for understanding the molecular and antigenic features of the virus. Based on the patterns of both in vivo and in vitro biological properties, rotavirus comprises seven distinct groups (A to G) and mature particles contains 11 segments of double-stranded RNA (dsRNA) genes which provide a suitable option for recombination such as effective intergroup reassortment or rearrangement of rotaviruses. Analysis of genomic electropherotypes is a relatively easy, rapid and popular technique for direct visualization of rotavirus genome and to monitor virus outbreaks and transmission in molecular epidemiology studies. In most cases, the electrophoretic pattern of group A rotaviruses genome is composed of four high-molecular-weight dsRNA segments (1 to 4), two middle-sized segments (5 and 6), a distinctive set of three segments (7 to 9), and two smaller segments (10 and 11) which are readily observed in polyacrylamide gel electrophoresis (PAGE) analysis on 10% gels to 1...

Research paper thumbnail of PCR Detection of Thymidine Kinase Gen of Latent Herpes Simplex Virus Type 1 in Mice Trigeminal Ganglia

archives of razi institute, 2006

Herpes simplex virus type 1 establishes a latent infection in the peripheral nervous system follo... more Herpes simplex virus type 1 establishes a latent infection in the peripheral nervous system following primary infection. During latent infection, virus genome exhibit limited transcription, with the HSV LATs consistently detected in latency infected ganaglia. Following ocular infection viral latency develops in the trigeminal ganglia. In this study PCR has been used for detection of HSV-1 nucliec acid . BALB/c mice were inoculated with HSV-1 and infected mice ganglia were applied to detect HSV-1 TK gene. Amplification of TK fragment in mice ganglia was considered as an indicator of viral latency. The results were suggested that TK PCR can be used as a reliable diagnostic tool and it is a specific and highly sensitive method for the detection of HSV genome in trigeminal ganglia.

Research paper thumbnail of Introduction of RNA virus evolution

Medical laboratory sciences, 2015

Lots of viruses, in particular RNA viruses, have high mutation rates and relatively short generat... more Lots of viruses, in particular RNA viruses, have high mutation rates and relatively short generation times. Particle stability during infection in nature or in laboratory triggers the evolutionary event toward different mechanisms such as genome segmentation, point mutation and recombination. The frequency of mutant genomes increase and modify the previous distribution, which, consequently, lead to emergence of a new infectious particle. Mutation and selection are the most fundamental processes in evolution. High mutation rate of RNA viruses has an important role in viral fitness. Therefore, it increase our understanding about molecular biology of viral infections and their evolution by selection, mutation could reliably determine our ability to challenge destructive viruses. This review focuses on existing impressions of genetic organization and mechanisms of RNA viruses evolution.

Research paper thumbnail of Survey of BK Virus in Renal Transplant Recipients in Iran: A Systematic Review and Meta-Analysis

Intervirology, 2020

Introduction: BK virus (BKV) infection in renal transplant (RT) recipients can cause hemorrhagic ... more Introduction: BK virus (BKV) infection in renal transplant (RT) recipients can cause hemorrhagic cystitis, transient renal dysfunction, and BKV nephropathy (BKVN). The prevalence and significance of BKV in RT recipients remain to be clarified in the Iranian population. The purpose of this review is to summarize the overall prevalence of BKV infection in RT recipients from previously published studies in Iran. Methods: We systematically reviewed articles through a comprehensive search of the main electronic and Persian national databases up to November 2019. Results: The overall pooled prevalence of BKV infection among the Iranian population was 23% (95% CI; 15–33%). Comparing these studies revealed that the prevalence of BKV in plasma samples ranges from 3 to 40%, in renal biopsies 1–13%, and in urine samples 10–49%. Due to substantial heterogeneity among reported studies (I2 = 93%, p < 0.01), random-effect meta-analysis was performed. BKV infection rate was slightly higher in wo...

Research paper thumbnail of Frequency of Cytomegalovirus Viral Load in Iranian Human Immunodeficiency Virus-1-Infected Patients with CD4+ Counts <100 Cells/mm3

Intervirology, 2021

Objectives: The aim of present work was to assess cytomegalovirus (CMV) viremia in Iranian human ... more Objectives: The aim of present work was to assess cytomegalovirus (CMV) viremia in Iranian human immunodeficiency virus (HIV)-1-infected patients with a CD4+ count <100 cells/mm3 and to explore whether CMV DNA loads correlate with CD4+ cell counts or associated retinitis. Methods: This study was conducted at the AIDS research center in Iran on HIV-1-infected patients with CD4+ count <100 cells/mm3, antiretroviral therapy-naive, aged ≥18 years with no previous history of CMV end-organ disease (CMV-EOD). Results: Thirty-nine of 82 patients (47.56%) had detectable CMV viral load ranging from 66 to 485,500 IU/mL. CMV viral load in patients with retinitis ranges from 352 to 2,720 IU/mL, and it was undetectable in 2 patients. No significant associations between CMV viremia and CD4+ cell count was found (p value = 0.31), whereas significant association of CMV viremia in HIV-infected patients with retinitis was found (p < 0.02). Conclusions: We estimated the frequency of CMV viral ...

Research paper thumbnail of Coxsackievirus B3 protease 3C induces cell death in eukaryotic cells

Iranian Journal of Virology, Jun 1, 2009

Coxsackievirus B3 (CVB3) is the most common agent known to cause viral myocarditis. The viral gen... more Coxsackievirus B3 (CVB3) is the most common agent known to cause viral myocarditis. The viral genome encodes a single polyprotein that is cleaved to produce several proteins by virally encoded proteases. Most of this proteolytic processing is catalyzed by a cysteine protease called 3C. The 3C protease plays major role in viral replication and cellular damage. To understand the mechanism of 3C function in virus infected cells and also development of antiviral agants against the virus, a 3C expressing plasmid was constructed. The cDNA of 3C protease was synthesized using CVB3 infected cells through reverse transcription process and was cloned in pcDNA3.1-. The constructed plasmid was confirmed by sequencing and restriction enzyme analysis. By transfection of the constructed plasmid into HeLa and MCF-7 cells, we showed that 3C protease induced cell death through multiple converging pathways, such as down regulation of cellular factors and decreasing of mRNA transcripts. This affect on HeLa cells as stronger than MCF-7 cells.

Research paper thumbnail of Autophagy induction regulates influenza virus replication in a time-dependent manner

Journal of medical microbiology, 2017

Autophagy plays a key role in host defence responses against microbial infections by promoting de... more Autophagy plays a key role in host defence responses against microbial infections by promoting degradation of pathogens and participating in acquired immunity. The interaction between autophagy and viruses is complex, and this pathway is hijacked by several viruses. Influenza virus (IV) interferes with autophagy through its replication and increases the accumulation of autophagosomes by blocking lysosome fusion. Thus, autophagy could be an effective area for antiviral research. In this study, we evaluated the effect of autophagy on IV replication. Two cell lines were transfected with Beclin-1 expression plasmid before (prophylactic approach) and after (therapeutic approach) IV inoculation.Results/Key findings. Beclin-1 overexpression in the cells infected by virus induced autophagy to 26 %. The log10haemagglutinin titre and TCID50 (tissue culture infective dose giving 50 % infection) of replicating virus were measured at 24 and 48 h post-infection. In the prophylactic approach, the ...

Research paper thumbnail of Protective Effects of DNA Vaccine Encoding Glycoprotein D-1 of Herpes Simplex Virus Type 1 on Latency Rate in Trigeminal Ganglia

The Neuroscience Journal of Shefaye Khatam, 2013

HSV-1 D-1 DNA TCID 5 × 10 5 (HSV-1) DNA D-1 D DNA DNA feature for vaccine development. Materials ... more HSV-1 D-1 DNA TCID 5 × 10 5 (HSV-1) DNA D-1 D DNA DNA feature for vaccine development. Materials & Methods protein D-1 (gD-1) gene of HSV-1 against lethal ocularly challenge of TCID 5 × 10 5 Results DNA vaccine encoding Herpes simplex virus Glycoprotein D gene on latency rate in TG and mortality rate. Conclusion

Research paper thumbnail of Determining influenza virus shedding at different time points in madin-darby canine kidney cell line

Cell journal, 2013

Monitoring of influenza virus shedding and optimization of multiplicities of infection (MOI) is i... more Monitoring of influenza virus shedding and optimization of multiplicities of infection (MOI) is important in the investigation of a virus one step growth cycle and for obtaining a high yield of virus in vaccine development and conventional basic diagnostic methods. However, eluted infectious viruses may still be present immediately after virus inoculation and when cells are washed following virus cultivation which may lead to a false positive virus infectivity assay. In this experimental study, we investigated influenza virus progeny production in Madin-Darby canine kidney (MDCK) cells with five different MOI at determined time points. The results were analyzed by end point titration tests and immunofluorescence assay. Higher titers of eluted virus were observed following a high MOI inoculation of virus in cell culture. Most probably, this was the result of sialic acid residues from viral hemagglutin in proteins that were cleaved by neuraminidase glycoproteins on the surface of the ...

Research paper thumbnail of Sequence analysis of the foot and mouth disease virus type O/IRN/2007 VP1 gene from Iranian isolate

Folia biologica, 2013

The foot and mouth disease virus (FMDV) causes a vesicular and contagious disease of cloven-hoofe... more The foot and mouth disease virus (FMDV) causes a vesicular and contagious disease of cloven-hoofed animals. In this study, the virus was isolated from vesicles of the infected cattle using cell culture and serotyped by ELISA test. The extracted RNA from the infected cells was reverse transcribed and amplified using VP1 gene-specific primer pairs by means of one-step RT-PCR. The purified VP1 gene was sub-cloned into the uniqe KpnI and BamHI cloning sites of the pcDNA3.1+ vector. The DH5α strain of E. coli was transformed by the vector. The sequences of sub-cloned FMDV type O/IRN/2007 VP1 were aligned with FMDV type O/UKG/2001 VP1 using MegAlign software. Nucleotide sequence comparisons were made using the BLAST software available from the NCBI website. The amino acid sequences of three sub-cloned FMDV type O/IRN/2007 VP1 were also aligned with three other similar sequences using MegAlign software. Nineteen of the most similar VP1 nucleotide sequences (by BLASTN program), FMDV O/IRN/2...

Research paper thumbnail of Development of ELISA test for determining the antibody titer against Herpes Simplex Virus-1 and comparison of the results by VNT

Research paper thumbnail of Rearranged Bovine Rotavirus Production through Cultivation of Virus by High Multiplicity of Infection (MOI) in Cell Culture and Amplification of Non-structural Genes using RT-PCR

Objective: Group A rotaviruses (GARV) are responsible for the vast majority of severe diarrhea wo... more Objective: Group A rotaviruses (GARV) are responsible for the vast majority of severe diarrhea worldwide that kills an estimated 600,000-870,000 children annually. Since infantile gastroenteritis is a main health problem, therefore diagnosis and treatment of this disease is crucial. Gene rearrangements have been detected in vitro during serial passages of the virus at a high multiplicity of infection (MOI) in cell culture, as well as in chronically infected immunodeficient individuals. In this study, we developed an RT-PCR method to detect and diagnose the standard and gene rearranged bovine rotavirus. Methods: Rotavirus RNA was extracted from confluent monolayers of infected MA-104 cells, stained with silver nitrate, and then electrophoresed in a 10% polyacrylamide gel. The full-length gene products that encoded the NSP1, 2, and 3 genes of the standard and rearranged rotavirus were amplified by RT-PCR using specific primers. Results: We observed rearranged NSP1 and NSP3 genes that had different migration patterns seen with polyacrylamide gel electrophoresis. NSP1, 2, and 3 gene segments from standard and rearranged rotaviruses were amplified by RT-PCR, then the complete nucleotide sequence of each gene was subjected to sequencing. The results showed the generation of gene rearrangement through serial passages of the bovine rotavirus RF strain. Conclusion: Serial passage of rotavirus in cell culture at a high MOI and chronic infection in immunodeficient target groups might alter rotavirus evolution. The methods utilized for detection and characterization of rotaviruses are continually evolving and being refined. Data collection is necessary to understand the molecular and antigenic features of the rotavirus in order to have a successful implementation of rotavirus studies and the development of a rotavirus vaccine. This study shows the importance of genetic variation and can provide valuable information about the amplification, diversity, biology, and evolution of rotaviruses.

Research paper thumbnail of Characterization and Transferring of Human Rotavirus Double-Layered Particles in MA104 Cells

Jundishapur Journal of Microbiology, 2014

Research paper thumbnail of Humoral immunogenicity assessment after receiving three types of SARS-CoV-2 vaccine

Scientific Reports

Several vaccines have been developed against SARS-CoV-2 and subsequently approved by national/int... more Several vaccines have been developed against SARS-CoV-2 and subsequently approved by national/international regulators. Detecting specific antibodies after vaccination enables us to evaluate the vaccine’s effectiveness. We conducted a prospective longitudinal study among members of Tarbiat Modares University of Tehran, Iran, from 4 September 2021 until 29 December 2021. We aimed to compare the humoral immunogenicity of 3 vaccine types. Participants consisted of 462 adults. Anti-SARS-CoV-2 receptor-binding domain [RBD] IgG titer was compared in 3 groups, each vaccinated by available vaccines in Iran at the time: Oxford/AstraZeneca, COVIran Barekat, and Sinopharm. The median IgG titer was: 91.2, 105.6, 224.0 BAU/ml for Sinopharm, COVIran Barekat and Oxford/AstraZeneca respectively after the first dose; 195.2, 192.0, 337.6 BAU/ml after the second one. We also analyzed the frequency of antibody presence in each vaccine group, in the same order the results were 59.0%, 62.6% and 89.4% aft...

Research paper thumbnail of Prevalence of herpes simplex viruses types 1 and 2 infections among suspected children of encephalitis in Kermanshah, Iran

Iranian Journal of Microbiology

Background and Objectives: Various infectious and non-infectious factors can cause encephalitis i... more Background and Objectives: Various infectious and non-infectious factors can cause encephalitis in the central nervous system (CNS), the most important of which are viruses. Herpes viruses are one of the most important causes of encephalitis worldwide. PCR detected the virus on the cerebrospinal fluid (CSF) sample. The aim of this study was to set up an in-house PCR to identify herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) and determine the prevalence of these viruses in suspected children of encephalitis. Materials and Methods: This cross-sectional study was conducted on 160 suspected children with encephalitis cases re- ferred to Dr. Kermanshahi Children Hospital, Kermanshah, Iran, from April to March 2021. CSF samples were extracted using a viral extraction kit, and a PCR was performed. The level of glucose and total protein of the samples was measured. Results: The total prevalence of HSV was 16.25%. 17 samples were positive for HSV-1 (10.6%), and 9...

Research paper thumbnail of The Effect of Oncolytic Reovirus Infection on Nitric Oxide Secretion and Induction of Apoptosis in Adipose Tissue-Derived Mesenchymal Stem Cells

Iranian Journal of Medical Microbiology

Background and Aims: Oncolytic viruses (OVs) are a new approach in treatment of cancer. Antitumor... more Background and Aims: Oncolytic viruses (OVs) are a new approach in treatment of cancer. Antitumor efficacy of OVs were limited due to insufficient and non-specific viral delivery to tumor sites. To overcome this issue, mesenchymal stem cells (MSCs) were used for their ability to specifically homing into tumors. The main aim of this study was to use MSCs as carriers and investigate the effect of oncolytic reovirus infection in MSCs, induction of apoptosis, nitric oxide (NO) secretion and their effects for selectively killing tumor cells, to use in future. Materials and Methods: MSCs isolated from mice adipose tissue and confirmed. Then, the ability of the virus to infect MSCs and the effect of reovirus infection in induction of apoptosis and NO secretion in MSCs were evaluated. Results: The results demonstrate that reovirus could replicate on MSCs. The finding indicated that the NO production significantly was higher at 72 h post infection with different MOI in comparison to the control cells. Also, reovirus induced high level of apoptosis in the MSCs at 48 h post infection compared with the control cells. Conclusions: Based on observed results, reovirus increased the secretion of iNOS (inducible nitric oxide) in the infected MSCs at 48 h post infection; therefore, high amounts of NO and reovirus replication were found to trigger apoptosis at 48 h post infection. Therefore, by optimizing the replication time of virus in the MSCs, specific viral delivery to tumor sites are available and causes cancer cells' death.

Research paper thumbnail of The Convenient and Economical Method to Collect Adipose Mesenchymal Stem Cells

Medical laboratory sciences, 2019

Background: Enzymatic digestion is an essential stage for culturing Mesenchymal Stem Cells (MSCs)... more Background: Enzymatic digestion is an essential stage for culturing Mesenchymal Stem Cells (MSCs) and their therapeutic application. Several factors such as being cost-benefit, efficiency, safety, yield and amount of produced cells are determinant for choosing the appropriate enzyme. Collagenase is a conventional enzyme commonly used for enzymatic digestion. However, other enzymes like trypsin and even combination of these enzymes can be used as an alternative strategy in different situations. Materials and Methods: Abdominal subcutaneous adipose tissue was obtained from male BALB/c mice and digested under three different enzymatic processes: collagenase and collagenase/trypsin and trypsin. Cell culture process was performed under standard condition and MSCs at 3rd passage were used for further characterization by flow cytometry. Results: In this study, two different enzymatic methods for digestion of adipose-derived MSCs (ADSCs) of BALB/c mice were investigated. The morphology of cells was pretty different and was more homogenous in collagenase group. Also the yield of cells was varied among groups. Furthermore, the obtained data from flow cytometry revealed that ADSCs were positive for CD90 (70%), CD29 (98%), CD105 (52%) and negative for CD45 (<2%).Conclusion: Application of different enzymes depends on various conditions. Altogether, these data indicate that although use of trypsin in isolation protocol is cost-benefit, it can be used as an alternative method whenever limited number of cells will be needed. However, collagenase as a well-known and conventional method can be used for isolation of larger quantity of cells with several applications.

Research paper thumbnail of The Effect of Autophagy Induction in Oncolytic Reovirus Replication in Mesenchymal Stem Cells

International Journal of Medical Laboratory, 2021

Background and Aims: Oncolytic reoviruses can infect and kill malignant cells while sparing their... more Background and Aims: Oncolytic reoviruses can infect and kill malignant cells while sparing their normal counterparts. Reoviral infection can induce or activate autophagy, even though metformin can induce autophagy. Identifying and regulating the cellular pathways important for reovirus replication and oncolysis can improve targeted-biological therapies for cancer. Here, the autophagic process was triggered via metformin, and we investigated the effect of autophagy activation on oncolytic reovirus replication in mesenchymal stem cells as primary cells and L929 cell lines. Materials and Methods: Adipose derived mesenchymal stem cells (AD-MSCs) and L929 cells were treated with metformin and reovirus type-3 strain Dearing (T3D). Twenty-four hours after infection, the viability of AD-MSCs and L929 cells were examined by MTT assay. Also, the effect of metformin-induced autophagy in the reovirus replication in these cells was determined by real-time polymerase-chain-reaction. Results: Our...

Research paper thumbnail of Development of a Western Blot Assay for Detection of Antibodies against HSV Using Purified HSV Virions Prepared by Sucrose Density Gradient

Objective(s) Herpes simplex viruses (HSVs) have widespread and ubiquitous prevalence in the human... more Objective(s) Herpes simplex viruses (HSVs) have widespread and ubiquitous prevalence in the human population and they have received a great deal of attention due to the range of diseases, they caused as a result of an infection. It seems that the fast and reliable diagnostic methods are needed for detecting the herpes simplex virus type 1 (HSV1) antibodies especially in patients with HSV encephalitis, immunocompromised people, and neonatal infections. The aim of this study was designing a Western blotting method for HSV1 antibody detection, using the purified virus by sucrose density gradient centrifugation procedure. Materials and Methods The most reliable method for HSV detection is virus neutralization test but it needs cell culture preparation, high expertise, as well as the high amounts of serum samples. Considering the difficulties of this method, we tried to run a new one for HSV antibody detection by propagating the viruses and then purify them by sucrose density gradient centrifugation method. The purified viruses used as antigens in Western blotting assay. Results Diluted sera (1:100, and 1:200 dilutions) used in Western blotting and twofold dilutions of the sera applied in virus neutralization test. Five of twenty seven samples were negative in Western blotting and the same results obtained in virus neutralization test. Comparing with our gold standard, the sensitivity and specificity of the developed assay were both 100%. Conclusion Our results show that the designed method is a reliable method for replacing the virus neutralization test in diagnostic laboratories. It can also, be used for confirming the ELISA results.

Research paper thumbnail of Kinetics of Oncolytic Reovirus T3D Replication and Growth Pattern in Mesenchymal Stem Cells

Cell Journal (Yakhteh), 2020

Objective Currently, application of oncolytic-virus in cancer treatment of clinical trials are gr... more Objective Currently, application of oncolytic-virus in cancer treatment of clinical trials are growing. Oncolytic-reovirus is an attractive anti-cancer therapeutic agent for clinical testing. Many studies used mesenchymal stem cells (MSCs) as a carrier cell to enhance the delivery and quality of treatment with oncolytic-virotherapy. But, biosynthetic capacity and behavior of cells in response to viral infections are different. The infecting process of reoviruses takes from two-hours to one-week, depends on host cell and the duration of different stages of virus replication cycle. The latter includes the binding of virus particle, entry, uncoating, assembly and release of progeny-viruses. We evaluated the timing and infection cycle of reovirus type-3 strain Dearing (T3D), using one-step replication experiment by molecular and conventional methods in MSCs and L929 cell as control. Materials and Methods In this experimental study, L929 and adipose-derived MSCs were infected with differ...

Research paper thumbnail of Development of Poly ( A )-Tailed Universal Reverse Transcription Rearranged Rotavirus

rotaviruses are necessary for understanding the molecular and antigenic features of the virus. Ba... more rotaviruses are necessary for understanding the molecular and antigenic features of the virus. Based on the patterns of both in vivo and in vitro biological properties, rotavirus comprises seven distinct groups (A to G) and mature particles contains 11 segments of double-stranded RNA (dsRNA) genes which provide a suitable option for recombination such as effective intergroup reassortment or rearrangement of rotaviruses. Analysis of genomic electropherotypes is a relatively easy, rapid and popular technique for direct visualization of rotavirus genome and to monitor virus outbreaks and transmission in molecular epidemiology studies. In most cases, the electrophoretic pattern of group A rotaviruses genome is composed of four high-molecular-weight dsRNA segments (1 to 4), two middle-sized segments (5 and 6), a distinctive set of three segments (7 to 9), and two smaller segments (10 and 11) which are readily observed in polyacrylamide gel electrophoresis (PAGE) analysis on 10% gels to 1...

Research paper thumbnail of PCR Detection of Thymidine Kinase Gen of Latent Herpes Simplex Virus Type 1 in Mice Trigeminal Ganglia

archives of razi institute, 2006

Herpes simplex virus type 1 establishes a latent infection in the peripheral nervous system follo... more Herpes simplex virus type 1 establishes a latent infection in the peripheral nervous system following primary infection. During latent infection, virus genome exhibit limited transcription, with the HSV LATs consistently detected in latency infected ganaglia. Following ocular infection viral latency develops in the trigeminal ganglia. In this study PCR has been used for detection of HSV-1 nucliec acid . BALB/c mice were inoculated with HSV-1 and infected mice ganglia were applied to detect HSV-1 TK gene. Amplification of TK fragment in mice ganglia was considered as an indicator of viral latency. The results were suggested that TK PCR can be used as a reliable diagnostic tool and it is a specific and highly sensitive method for the detection of HSV genome in trigeminal ganglia.

Research paper thumbnail of Introduction of RNA virus evolution

Medical laboratory sciences, 2015

Lots of viruses, in particular RNA viruses, have high mutation rates and relatively short generat... more Lots of viruses, in particular RNA viruses, have high mutation rates and relatively short generation times. Particle stability during infection in nature or in laboratory triggers the evolutionary event toward different mechanisms such as genome segmentation, point mutation and recombination. The frequency of mutant genomes increase and modify the previous distribution, which, consequently, lead to emergence of a new infectious particle. Mutation and selection are the most fundamental processes in evolution. High mutation rate of RNA viruses has an important role in viral fitness. Therefore, it increase our understanding about molecular biology of viral infections and their evolution by selection, mutation could reliably determine our ability to challenge destructive viruses. This review focuses on existing impressions of genetic organization and mechanisms of RNA viruses evolution.

Research paper thumbnail of Survey of BK Virus in Renal Transplant Recipients in Iran: A Systematic Review and Meta-Analysis

Intervirology, 2020

Introduction: BK virus (BKV) infection in renal transplant (RT) recipients can cause hemorrhagic ... more Introduction: BK virus (BKV) infection in renal transplant (RT) recipients can cause hemorrhagic cystitis, transient renal dysfunction, and BKV nephropathy (BKVN). The prevalence and significance of BKV in RT recipients remain to be clarified in the Iranian population. The purpose of this review is to summarize the overall prevalence of BKV infection in RT recipients from previously published studies in Iran. Methods: We systematically reviewed articles through a comprehensive search of the main electronic and Persian national databases up to November 2019. Results: The overall pooled prevalence of BKV infection among the Iranian population was 23% (95% CI; 15–33%). Comparing these studies revealed that the prevalence of BKV in plasma samples ranges from 3 to 40%, in renal biopsies 1–13%, and in urine samples 10–49%. Due to substantial heterogeneity among reported studies (I2 = 93%, p < 0.01), random-effect meta-analysis was performed. BKV infection rate was slightly higher in wo...

Research paper thumbnail of Frequency of Cytomegalovirus Viral Load in Iranian Human Immunodeficiency Virus-1-Infected Patients with CD4+ Counts <100 Cells/mm3

Intervirology, 2021

Objectives: The aim of present work was to assess cytomegalovirus (CMV) viremia in Iranian human ... more Objectives: The aim of present work was to assess cytomegalovirus (CMV) viremia in Iranian human immunodeficiency virus (HIV)-1-infected patients with a CD4+ count <100 cells/mm3 and to explore whether CMV DNA loads correlate with CD4+ cell counts or associated retinitis. Methods: This study was conducted at the AIDS research center in Iran on HIV-1-infected patients with CD4+ count <100 cells/mm3, antiretroviral therapy-naive, aged ≥18 years with no previous history of CMV end-organ disease (CMV-EOD). Results: Thirty-nine of 82 patients (47.56%) had detectable CMV viral load ranging from 66 to 485,500 IU/mL. CMV viral load in patients with retinitis ranges from 352 to 2,720 IU/mL, and it was undetectable in 2 patients. No significant associations between CMV viremia and CD4+ cell count was found (p value = 0.31), whereas significant association of CMV viremia in HIV-infected patients with retinitis was found (p < 0.02). Conclusions: We estimated the frequency of CMV viral ...

Research paper thumbnail of Coxsackievirus B3 protease 3C induces cell death in eukaryotic cells

Iranian Journal of Virology, Jun 1, 2009

Coxsackievirus B3 (CVB3) is the most common agent known to cause viral myocarditis. The viral gen... more Coxsackievirus B3 (CVB3) is the most common agent known to cause viral myocarditis. The viral genome encodes a single polyprotein that is cleaved to produce several proteins by virally encoded proteases. Most of this proteolytic processing is catalyzed by a cysteine protease called 3C. The 3C protease plays major role in viral replication and cellular damage. To understand the mechanism of 3C function in virus infected cells and also development of antiviral agants against the virus, a 3C expressing plasmid was constructed. The cDNA of 3C protease was synthesized using CVB3 infected cells through reverse transcription process and was cloned in pcDNA3.1-. The constructed plasmid was confirmed by sequencing and restriction enzyme analysis. By transfection of the constructed plasmid into HeLa and MCF-7 cells, we showed that 3C protease induced cell death through multiple converging pathways, such as down regulation of cellular factors and decreasing of mRNA transcripts. This affect on HeLa cells as stronger than MCF-7 cells.

Research paper thumbnail of Autophagy induction regulates influenza virus replication in a time-dependent manner

Journal of medical microbiology, 2017

Autophagy plays a key role in host defence responses against microbial infections by promoting de... more Autophagy plays a key role in host defence responses against microbial infections by promoting degradation of pathogens and participating in acquired immunity. The interaction between autophagy and viruses is complex, and this pathway is hijacked by several viruses. Influenza virus (IV) interferes with autophagy through its replication and increases the accumulation of autophagosomes by blocking lysosome fusion. Thus, autophagy could be an effective area for antiviral research. In this study, we evaluated the effect of autophagy on IV replication. Two cell lines were transfected with Beclin-1 expression plasmid before (prophylactic approach) and after (therapeutic approach) IV inoculation.Results/Key findings. Beclin-1 overexpression in the cells infected by virus induced autophagy to 26 %. The log10haemagglutinin titre and TCID50 (tissue culture infective dose giving 50 % infection) of replicating virus were measured at 24 and 48 h post-infection. In the prophylactic approach, the ...

Research paper thumbnail of Protective Effects of DNA Vaccine Encoding Glycoprotein D-1 of Herpes Simplex Virus Type 1 on Latency Rate in Trigeminal Ganglia

The Neuroscience Journal of Shefaye Khatam, 2013

HSV-1 D-1 DNA TCID 5 × 10 5 (HSV-1) DNA D-1 D DNA DNA feature for vaccine development. Materials ... more HSV-1 D-1 DNA TCID 5 × 10 5 (HSV-1) DNA D-1 D DNA DNA feature for vaccine development. Materials & Methods protein D-1 (gD-1) gene of HSV-1 against lethal ocularly challenge of TCID 5 × 10 5 Results DNA vaccine encoding Herpes simplex virus Glycoprotein D gene on latency rate in TG and mortality rate. Conclusion

Research paper thumbnail of Determining influenza virus shedding at different time points in madin-darby canine kidney cell line

Cell journal, 2013

Monitoring of influenza virus shedding and optimization of multiplicities of infection (MOI) is i... more Monitoring of influenza virus shedding and optimization of multiplicities of infection (MOI) is important in the investigation of a virus one step growth cycle and for obtaining a high yield of virus in vaccine development and conventional basic diagnostic methods. However, eluted infectious viruses may still be present immediately after virus inoculation and when cells are washed following virus cultivation which may lead to a false positive virus infectivity assay. In this experimental study, we investigated influenza virus progeny production in Madin-Darby canine kidney (MDCK) cells with five different MOI at determined time points. The results were analyzed by end point titration tests and immunofluorescence assay. Higher titers of eluted virus were observed following a high MOI inoculation of virus in cell culture. Most probably, this was the result of sialic acid residues from viral hemagglutin in proteins that were cleaved by neuraminidase glycoproteins on the surface of the ...

Research paper thumbnail of Sequence analysis of the foot and mouth disease virus type O/IRN/2007 VP1 gene from Iranian isolate

Folia biologica, 2013

The foot and mouth disease virus (FMDV) causes a vesicular and contagious disease of cloven-hoofe... more The foot and mouth disease virus (FMDV) causes a vesicular and contagious disease of cloven-hoofed animals. In this study, the virus was isolated from vesicles of the infected cattle using cell culture and serotyped by ELISA test. The extracted RNA from the infected cells was reverse transcribed and amplified using VP1 gene-specific primer pairs by means of one-step RT-PCR. The purified VP1 gene was sub-cloned into the uniqe KpnI and BamHI cloning sites of the pcDNA3.1+ vector. The DH5α strain of E. coli was transformed by the vector. The sequences of sub-cloned FMDV type O/IRN/2007 VP1 were aligned with FMDV type O/UKG/2001 VP1 using MegAlign software. Nucleotide sequence comparisons were made using the BLAST software available from the NCBI website. The amino acid sequences of three sub-cloned FMDV type O/IRN/2007 VP1 were also aligned with three other similar sequences using MegAlign software. Nineteen of the most similar VP1 nucleotide sequences (by BLASTN program), FMDV O/IRN/2...

Research paper thumbnail of Development of ELISA test for determining the antibody titer against Herpes Simplex Virus-1 and comparison of the results by VNT

Research paper thumbnail of Rearranged Bovine Rotavirus Production through Cultivation of Virus by High Multiplicity of Infection (MOI) in Cell Culture and Amplification of Non-structural Genes using RT-PCR

Objective: Group A rotaviruses (GARV) are responsible for the vast majority of severe diarrhea wo... more Objective: Group A rotaviruses (GARV) are responsible for the vast majority of severe diarrhea worldwide that kills an estimated 600,000-870,000 children annually. Since infantile gastroenteritis is a main health problem, therefore diagnosis and treatment of this disease is crucial. Gene rearrangements have been detected in vitro during serial passages of the virus at a high multiplicity of infection (MOI) in cell culture, as well as in chronically infected immunodeficient individuals. In this study, we developed an RT-PCR method to detect and diagnose the standard and gene rearranged bovine rotavirus. Methods: Rotavirus RNA was extracted from confluent monolayers of infected MA-104 cells, stained with silver nitrate, and then electrophoresed in a 10% polyacrylamide gel. The full-length gene products that encoded the NSP1, 2, and 3 genes of the standard and rearranged rotavirus were amplified by RT-PCR using specific primers. Results: We observed rearranged NSP1 and NSP3 genes that had different migration patterns seen with polyacrylamide gel electrophoresis. NSP1, 2, and 3 gene segments from standard and rearranged rotaviruses were amplified by RT-PCR, then the complete nucleotide sequence of each gene was subjected to sequencing. The results showed the generation of gene rearrangement through serial passages of the bovine rotavirus RF strain. Conclusion: Serial passage of rotavirus in cell culture at a high MOI and chronic infection in immunodeficient target groups might alter rotavirus evolution. The methods utilized for detection and characterization of rotaviruses are continually evolving and being refined. Data collection is necessary to understand the molecular and antigenic features of the rotavirus in order to have a successful implementation of rotavirus studies and the development of a rotavirus vaccine. This study shows the importance of genetic variation and can provide valuable information about the amplification, diversity, biology, and evolution of rotaviruses.

Research paper thumbnail of Characterization and Transferring of Human Rotavirus Double-Layered Particles in MA104 Cells

Jundishapur Journal of Microbiology, 2014