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Background: There are numerous human genes associated with viral infections, and their identification in specific populations can provide suitable therapeutic targets for modulating the host immune system response and better understanding the viral pathogenic mechanisms. Many antiviral signaling pathways, including Type I interferon and NF-κB, are regulated by TRIM proteins. Therefore, the identification of TRIM proteins involved in COVID-19 infection can play a significant role in understanding the innate immune response to this virus. Methods: In this study, the expression of TRIM25 gene was evaluated in a blood sample of 330 patients admitted to the hospital (142 patients with severe disease and 188 patients with mild disease) as well as in 160 healthy individuals. The relationship between its expression and the severity of COVID-19 disease was assessed and compared among the study groups by quantitative Real-time PCR technique. The statistical analysis of the results demonstrated a significant reduction in the expression of TRIM25 in the group of patients with severe infection compared to those with mild infection. Furthermore, the impact of increased expression of TRIM25 gene in HEK-293 T cell culture was investigated on the replication of attenuated SARS-CoV-2 virus. Results: The results of Real-time PCR, Western blot for the viral nucleocapsid gene of virus, and CCID50 test indicated a decrease in virus replication in these cells. The findings of this research indicated that the reduced expression of the TRIM25 gene was associated with increased disease severity of COVID-19 in individuals. Additionally, the results suggested the overexpression of TRIM25 gene can impress the replication of attenuated SARS-CoV-2 and the induction of beta-interferon. Conclusion: TRIM25 plays a critical role in controlling viral replication through its direct interaction with the virus and its involvement in inducing interferon during the early stages of infection. This makes TRIM25 a promising target for potential therapeutic interventions.

ObjectiveSpirulina (arthrospira platensis) is a cyanobacterium proven to have anti-inflammatory, antiviral, and antioxidant effects. However, the effect of high-dose Spirulina supplementation on hospitalized adults with COVID-19 is currently unclear. This study aimed to evaluate the efficacy and safety of high-dose Spirulina platensis for SARS-CoV-2 infection.Study DesignWe conducted a randomized, controlled, open-label trial involving 189 patients with COVID-19 who were randomly assigned in a 1:1 ratio to an experimental group that received 15.2g of Spirulina supplement plus standard treatment (44 non-intensive care unit (non-ICU) and 47 ICU), or to a control group that received standard treatment alone (46 non-ICU and 52 ICU). The study was conducted over six days. Immune mediators were monitored on days 1, 3, 5, and 7. The primary outcome of this study was mortality or hospital discharge within seven days, while the overall discharge or mortality was considered the secondary outcome.ResultsWithin seven days, there were no deaths in the Spirulina group, while 15 deaths (15.3%) occurred in the control group. Moreover, within seven days, there was a greater number of patients discharged in the Spirulina group (97.7%) in non-ICU compared to the control group (39.1%) (HR, 6.52; 95% CI, 3.50 to 12.17). Overall mortality was higher in the control group (8.7% non-ICU, 28.8% ICU) compared to the Spirulina group (non-ICU HR, 0.13; 95% CI, 0.02 to 0.97; ICU, HR, 0.16; 95% CI, 0.05 to 0.48). In non-ICU, patients who received Spirulina showed a significant reduction in the levels of IL-6, TNF-α, IL-10, and IP-10 as intervention time increased. Furthermore, in ICU, patients who received Spirulina showed a significant decrease in the levels of MIP-1α and IL-6. IFN-γ levels were significantly higher in the intervention group in both ICU and non-ICU subgroups as intervention time increased. No side effects related to Spirulina supplements were observed during the trial.ConclusionHigh-dose Spirulina supplements coupled with the standard treatment of COVID-19 may improve recovery and remarkably reduce mortality in hospitalized patients with COVID-19.Clinical Trial Registrationhttps://irct.ir/trial/54375, Iranian Registry of Clinical Trials number (IRCT20210216050373N1)

Abstract Background Tripartite motif-containing 28 (TRIM28) is an impressive regulator of the epigenetic control of the antiviral immune response. This study evaluated if the differential expression of TRIM28 correlates with the severity of coronavirus disease 2019 (COVID-19) infection. Methods A total of 330 COVID-19 patients, including 188 mild and 142 severe infections, and 160 healthy controls were enrolled in this study. Quantitative real-time polymerase chain reaction (qPCR) was used to determine the expression levels of TRIM28 in the studied patients. Results TRIM28 mRNA levels were significantly lower in both groups of patients versus the control group and in the severe group indicated further reduction in comparison to mild infection. The multivariate logistic regression analysis showed the mean age, lower levels of low-density lipoprotein (LDL), high-density lipoprotein (HDL), cholesterol, lower 25-hydroxyvitamin D, and PCR cycle threshold (Ct) value and higher levels of erythrocyte sedimentation rate (ESR) and differential expression of TRIM28 were linked to the severity of COVID-19 infection. Conclusion The results of this study proved that the downregulation of TRIM28 might be associated with the severity of COVID-19 infection. Further studies are required to determine the association between the COVID-19 infection severity and TRIM family proteins.

Abstract Background The recent pandemic caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has elevated several clinical and scientific questions. These include how host genetic factors influence the pathogenesis and disease susceptibility. Therefore, the aim of this study was to evaluate the impact of interferon lambda 3 and 4 (IFNL3/4) gene polymorphisms and clinical parameters on the resistance and susceptibility to coronavirus disease 2019 (COVID-19) infection. Methods A total of 750 SARS-CoV-2 positive patients (375 survivors and 375 nonsurvivors) were included in this study. All single-nucleotide polymorphisms (SNPs) on IFNL3 (rs12979860, rs8099917, and rs12980275) and IFNL4 rs368234815 were genotyped by the polymerase chain reaction-restriction fragment length polymorphism (PCR–RFLP) method. Results In this study, a higher viral load (low PCR Ct value) was shown in nonsurvivor patients. In survivor patients, the frequency of the favorable genotypes of IFNL3/4 SNPs (rs12979860 CC, rs12980275 AA, rs8099917 TT, and rs368234815 TT/TT) was significantly higher than in nonsurvivor patients. Multivariate logistic regression analysis has shown that a higher low-density lipoprotein (LDL), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and PCR Ct value, and lower 25-hydroxyvitamin D, and also IFNL3 rs12979860 TT, IFNL3 rs8099917 GG, IFNL3 rs12980275 GG, and IFNL4 rs368234815 ∆G/∆G genotypes were associated with the severity of COVID-19 infection. Conclusions The results of this study proved that the severity of COVID-19 infection was associated with clinical parameters and unfavorable genotypes of IFNL3/IFNL4 SNPs. Further studies in different parts of the world are needed to show the relationship between severity of COVID-19 infection and host genetic factors.

Abstract Infection with human immunodeficiency virus (HIV)‐1 causes immunological disorders and death worldwide which needs to be further assisted by novel anti‐retroviral drug delivery systems. Consequently, finding newer anti‐retroviral pharmaceuticals by using biocompatible, biodegradable nanomaterials comprising a nanoparticle as core and a therapeutic agent is of high global interest. In this experiment, a second generation of a negatively charged nano‐biopolymer linear globular G2 dendrimer was carefully conjugated and loaded with well‐known anti‐HIV drugs lamivudine and efavirenz, respectively. They were characterised by a variety of analytical methods such as Zetasizer, Fourier‐transform infrared spectroscopy, elemental analysis and liquid chromatography‐mass spectroscopy. Additionally, conjugated lamivudine and loaded efazirenz with globular PEGylated G2 dendrimer were tested on an HEK293 T cell infected by single‐cycle replicable HIV‐1 virion and evaluated using XTT test and HIV‐1 P24 protein load. The results showed that lamivudine‐conjugated G2 significantly decreased retroviral activity without any cell toxicity. This effect was more or less observed by efavirenz‐loaded G2. These nano‐constructs are strongly suggested for further in vivo anti‐HIV assays.

Hepatitis B and C viruses are major public health problems. These viruses can chronically lead to liver disease such as fibrosis, cirrhosis, and hepatocellular carcinoma, which often increase mortality in these patients. According to previous studies, the liver is highly affected by changes in the microbiota of gastrointestinal tract and immune system damage caused by inflammation due to viral hepatitis. Significant advances have been made in identifying gastrointestinal microbiota in cirrhotic patients associated with viral hepatitis and its use in their prognosis and treatment in recent years. Unique bacterial profiles are observed in cirrhotic patients associated with viral hepatitis, including increased numbers of Streptococcus, Prevotella, Staphylococcus, and Enterococcus, as well as decreased numbers of Clostridium and Ruminococcus. The purpose of this review was to summarize and discuss the gastrointestinal microbiota profile in patients with chronic hepatitis B and C and its role in the progression of cirrhosis.

Non-responsiveness or poor responsiveness to vaccines are challenging issues in vaccine development, and efforts have been made to find out the potential reasons for these conditions. Intestinal microbiome plays a key role in regulating and development of immune system and the composition and diversity of microbiota in different individuals on the one hand, and the imbalance of intestinal microbial population (Dysbiosis) on the other hand, could be considered as the most effective factors on the human immune system’s response to vaccines. Herein, we reviewed studies on the relationships between gut microbiome and immune response to vaccines. It is known that higher relative abundance of Actinobacteria and Firmicutes could induce more effective immune response to vaccines. In contrast, higher relative frequencies of Proteobacteria and Bacteroidetes are reported in weak immune response to vaccines. There is a strong interaction between the development of immune system and the composition of microbiota throughout life, therefore, it is important to determine the best times for examinations and using the most accurate methods, including sequencing the entire genome and Next Generation Sequencing (NGS). However, interpretation of the results of these researches depends on study designs, inclusion/exclusion criteria, timing of sampling according to the time of vaccination, and the methods used in each study. These considerations could lead to more reliable results and provide better understanding on the crosstalk between microbiota and immune response to vaccines, which will consequently improve vaccine efficacy.

Introduction: Hepatitis E virus (HEV) causes emerging diseases in poor regions of the world. The ORF2 is the only protein encoded by the virus to make the viral capsid. The aggregation of proteins into inclusion bodies (IBs) while expressing ORF2 is a major challenge in bioengineering. Methods: The ORF2 conserved sequence was expressed in Escherichia coli BL21 and assessed by a modified SDS-PAGE containing a weak denaturing environment to solubilize the recombinant ORF2 protein and Western blotting. The protein of interest was evaluated by secondary structure prediction using SOPMA, homology modeling by I-TASSER and Circular Dichroism analyses. The function of the recombinant protein was investigated by an in-house ELISA using serum specimens of HEV infected patients. Results: The solubilized form of ORF2 protein was successfully expressed in E. coli BL21 and was confirmed by SDS-PAGE and Western blotting. Secondary structure prediction, homology modeling and CD analysis of the protein of interest demonstrated that the native structure of ORF2 was almost intact. The specific anti-HEV antibody was detected using this recombinant protein and an in-house ELISA test. Conclusion: We achieved new combinations of chemical agents, consisted of low concentrations of urea and detergents to overcome the aggregation of ORF2 protein in IBs inside E. coli BL21.

Introduction: To date, while many studies have investigated antiviral agents or vaccines against HIV, success has been limited. In this field, mutagenesis of the viral genome often contributes to viral escape from the antiretroviral therapies and the emergence of HIV-1 strains resistant to the drugs. Therefore, developing alternative methods, including more effective vaccines, antiviral therapies (such as RNAi therapy) and delivery systems, seems to be necessary to compensate for these issues. The aim of this study was to establish an efficient system for siRNA delivery as a safe anti-HIV therapeutic approach. Methods: In this research, the use of chitosan-coated SPION was investigated as a method for RNA delivery. So, after generating a stable cell line of HEK293 (expressing HIV-1 tat), we designed a potent siRNA against HIV-1 tat, and then the effectiveness of the modified SPION in siRNA delivery to HEK293 cells was evaluated. Results: The results of this study showed that the optimal concentration (50 µg/mL) of the modified SPION containing anti-tat siRNA (with a range size of 50-70 nm and average zeta potential of +25 mV) were significantly internalized into the cells and decreased the expression of HIV-1 tat over than 80%. Moreover, the nanoparticles showed no considerable toxicity on the cells. Conclusion: It would be concluded that SPION could be optimized as a probable RNA/vaccine delivery system into target cells. Therefore, this study offers a therapeutic strategy against HIV or other infectious diseases.

BackgroundHepatitis C virus (HCV), non-structural 5A (NS5A), and non-structural 5B (NS5B) resistance-associated substitutions (RASs) are the main causes of failure to direct-acting antiviral agents (DAAs). NS5A and NS5B RASs can occur in patients with HCV infection naturally and before exposure to DAAs.ObjectivesThis study aimed to evaluate naturally-occurring NS5A and NS5B RASs in Iranian patients with HCV genotype 1a (HCV-1a) and -3a infections.MethodsIn this cross-sectional study, viral RNA was extracted from serum specimens. NS5A and NS5B regions were amplified using RT-PCR followed by DNA sequencing. The results of nucleotide sequences were aligned against reference sequences of HCV-1a and -3a and the amino acid substitutions were analyzed using geno2pheno [hcv] web application.ResultsAmong 135 patients with hepatitis C, NS5A amino acid substitutions/RASs were identified in 26.4% and 15.9% of patients with HCV-1a and -3a infections, respectively. The identified amino acid substitutions/RASs in the NS5A region of patients with HCV-1a infection were M28T/V/I 11.1%, Q30R/H 4.2%, L31M 1.4%, and H58Y/P/C/D/Q/S/T 16.7%. Y93H substitution was not found in HCV-1a sequences. In patients with HCV-3a infection, NS5A amino acid substitutions/RASs were A30T/K 9.5%, L31F 1.6%, P58S/T/C 3.2%, Y93H 3.2%, and Y93N 3.2%. No resistance substitutions were identified in NS5B sequences from patients with HCV-1a and -3a infections.ConclusionIn this study, baseline amino acid substitutions/RASs were only identified in the NS5A region in Iranian patients with HCV-1a and -3a infections, and the prevalence of these amino acid substitutions/RASs were in accordance with similar studies. There were no RASs in the HCV-1a and -3a NS5B region.

Introduction : Viral meningitis is an inflammation of the leptomeninges as a manifestation of central nervous system (CNS) infection, and more than 85% of viral meningitis cases are caused by non-polio enteroviruses. Molecular methods such as polymerase chain reaction (PCR) are increasingly used to detect and type non-polio enteroviruses. Here, we describe the impact of these viruses on aseptic meningitis in young children in Iran. Methods: Cerebrospinal fluids were collected from 366 children under 8 years old with primary clinical diagnosis of aseptic meningitis. Real-Time RT-PCR and PCR were performed using specific primers for each human enterovirus (HEV), human parechovirus (HPeV), and enterovirus 71 (EV71). Results: Positive results were obtained for HEV, HPeV, and EV71 in 118 (32.2%), 154 (42%), and 94 (25.7%) patients, respectively. HPeV detection rate was higher in both male and female patients with no statistically significant difference. Conclusion: Using molecular methods, such as PCR as routine laboratory diagnostic tests for HEVs will result in better understanding of disease burden of these viruses, especially in central nervous system involvements, and will consequently reduce hospitalization and use of antibiotic, which often lead to other complications. J Med Microbiol Infec Dis, 2014, 2 (2): 5 pages.

Background: Despite the effective pre- and post-exposure treatments, at least 60,000 deaths from rabies occur worldwide every year. Post-exposure treatment is considered as one of the most significant measures for preventing human deaths in exposed individuals. The 2-1-1 rabies post-exposure treatment schedule known as Zagreb regimen is an abbreviated immunization plan in which a tissue culture rabies vaccine is administered intramuscularly at two sites on day 0 and at one site on days 7 and 21. Objectives: In this study the efficacy of rabies vaccine administration under Zagreb regimen in an Iranian group of patients attending Pasteur Institute of Iran was evaluated. Methods: Rabies neutralizing antibody titer was measured in 75 serum samples collected from 25 volunteers receiving post exposure treatment by rapid fluorescent focus inhibition test (RFFIT), and ELISA. Results: All patients were negative for rabies antibody in both ELISA and FRRIT tests on day 0 . A satisfactory rabies virus antibody response with the titer of ≥0.5 IU/ml was detected in all patients on day 21 and two weeks after the completion of vaccination (day 35). Conclusions: Rabies immunization with rabies Vero-cell vaccine (PVRV) under the 2-1-1 schedule (Zagreb regimen) could result in an adequate immune response without any adverse effect. However a more comprehensive comparative study is underway to confirm these findings . Vac Res , 2014, 1 (1): 26-28

Background and Objectives: Hepatitis C virus (HCV) chronically infects around 200 million people worldwide and fre- quently causes liver cirrhosis and hepatocellular carcinoma. Rapid detection of this virus results in decreasing the distance between infection and initiation the anti-viral treatment, and may prevent most of the undesirable consequences. The new detected HCV protein "Core+1" made from the ribosomal frame shift in Core region is an important candidate for diagnostic tools. This study was conducted to design a recombinant Bacmid plasmid expressing the HCV 1a Core+1 sequence in the Baculovirus expression system for further diagnostic applications. Materials and Methods: The HCV Core +1 gene was amplified by PCR using the pcDNA-HAF recombinant vector that contained the Core+1 sequence from HCV genotype 1a as a template, and the specific primers with 2 restriction sites for Nco I and Xba I restriction enzymes. The PCR product was cloned in XbaI/NcoI restriction sites of the linearized pFastBac-HTB vector and evaluated by using those restriction enzymes and sequencing. Then the recombinant pFastBac-HTB vector was transformed in DH10Bac and the result was screened and confirmed by X-Gal discrimination and PCR. Results: The HCV 1a Core+1 was successfully amplified and the PCR product was confirmed by using the related restriction enzymes and sequencing. Cloning of pFastBac vector with the purified PCR product of HCV Core+1 was confirmed. Finally, the recombinant Bacmid was successfully transformed in DH10Bac. Conclusion: The recombinant Bac-Core+1 expression vector is considered as an important tool to transfect the sf9 cell line and expression the Core+1 protein.

Post exposure prophylaxis using one of the WHO-approved vaccines is the method of choice for preventing rabies. Abnormal immune function in patients with some specific medical conditions, such as pregnancy, chronic hepatitis B virus infection, different types of cancers like lymphoma, diabetes I and II, corticosteroid consumption by patients with rheumatoid arthritis and lupus erythematosus, could impair the immunologic response to various vaccines. The immune response to rabies vaccination has never been examined in patients with any of these described medical conditions. This study purposed to evaluate the neutralyzing antibody response after vaccination with purified Vero cell rabies vaccine (PVRV) according to the WHO-recommended Post-Exposure Prophylaxis (PEP) "ESSEN" regimen.Thirty healthy volunteers and 50 volunteers with different medical conditions who were exposed to a suspected rabid animal in the 2nd or 3rd category of exposure received 5 doses of PVRV under the ESSEN protocol. Three blood samples were collected on days 0 (before the first dose), 14, and 35. The anti-rabies antibody titer was measured using the Rapid Fluorescent Foci Inhibition Test (RFFIT) and an ELISA Bio-Rad, Platelia, Rabies II kit.All subjects reached NAb titers above 0.5 IU/ml by day 14 after vaccination. On day 35 (1 week after receiving the last rabies vaccine), anti-rabies antibodies were in the protective level (>0.5 IU/ml) in both groups. There was no statistically significant difference in anti-rabies antibody response due to the type of exposure (category 2 or 3), and successful seroconversion was confirmed in both groups.In conclusion, the ESSEN protocol using the PVRV vaccine is sufficient for rabies prophylaxis in patients with specific medical conditions.

In 2008, African swine fever was introduced into Georgia, after which it spread to neighboring Armenia, Azerbaijan, and the Russian Federation. That same year, PCR and sequence analysis identified African swine fever virus in samples from 3 dead female wild boars in northwestern Iran. Wild boars may serve as a reservoir.