Mutational dynamics of SARS-CoV-2: Impact on future COVID-19 vaccine strategies.

The rapid emergence of multiple strains of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) has sparked profound concerns regarding the ongoing evolution of the virus and its potential impact on global health. Classified by the World Health Organization (WHO) as variants of concern (VOC), these strains exhibit heightened transmissibility and pathogenicity, posing significant challenges to existing vaccine strategies. Despite widespread vaccination efforts, the continual evolution of SARS-CoV-2 variants presents a formidable obstacle to achieving herd immunity. Of particular concern is the coronavirus spike (S) protein, a pivotal viral surface protein crucial for host cell entry and infectivity. Mutations within the S protein have been shown to enhance transmissibility and confer resistance to antibody-mediated neutralization, undermining the efficacy of traditional vaccine platforms. Moreover, the S protein undergoes rapid molecular evolution under selective immune pressure, leading to the emergence of diverse variants with distinct mutation profiles. This review underscores the urgent need for vigilance and adaptation in vaccine development efforts to combat the evolving landscape of SARS-CoV-2 mutations and ensure the long-term effectiveness of global immunization campaigns.

Assessment of species distribution and virulence factors of oral fungal carriage among hospitalized patients with COVID-19: a case-control study.

Background: The COVID-19 pandemic highlighted the need to study oral fungal carriage and its potential impact. In oral fungal environments, factors like changes in respiratory epithelium, increased pathogen attachment, local inflammation, and virulence factors could influence COVID-19 severity. The authors conducted a study to explore oral fungal carriage in COVID-19 patients and compare it to a healthy control group. Methods: The authors executed a case-control investigation including 144 COVID-19 patients and an equivalent number of 144 healthy controls. The matching criteria encompassed age, sex, body mass index, and the history of antibiotic and antiviral medication intake. This research was performed over a span of 12 months from May 2021 to May 2022. The mouth area was sampled with a cotton-tipped swab. Subsequently, all the samples underwent fungal culture and PCR-sequencing procedures. Results: In COVID-19 patients, oral fungal carriage was three times higher compared to healthy controls. Candida was the exclusive genus found in both groups, with Candida albicans being the most frequently isolated species (90.79%). Among COVID-19 patients, Candida species showed significantly higher esterase, proteinase, and hemolysin activity compared to healthy individuals. Both groups exhibited elevated levels of C. albicans virulence factors compared to non-albicans species. Conclusions: It is crucial to understand the way that virulence factors of oral fungal carriage act in COVID-19 patients in order to come up with novel antifungal medications, identify the contributing factors to drug resistance, and manage clinical outcomes.

Targeting CD47 as a therapeutic strategy: A common bridge in the therapy of COVID-19-related cancers.

Macrophages are essential mediators of innate immunity. Non-self-cells resist phagocytosis through the expression of the checkpoint molecule CD47. CD47, as the integrin-associated protein, is overexpressed on tumor and SARS-CoV-2-infected cells as a potential surface biomarker for immune surveillance evasion. CD47-signal-regulating protein alpha (SIRPα) interaction is a promising innate immunotarget. Previous findings based on monoclonal antibodies (mAbs) or fusion proteins that block CD47 or SIRPα have been developed in cancer research. While CD47 efficacy in infectious diseases, especially severe COVID-19 studies, is lacking, focus on macrophage-mediated immunotherapy that increases "eat me" signals in combination therapy with mAbs is optimistic. This integrin-related protein can be as a potential target to therapy for COVID-19. Here, we concentrate on the role of the CD47 signaling pathway as a novel therapeutic strategy for COVID-19-associated cancer treatment.

Alzheimer's disease and microorganisms: the non-coding RNAs crosstalk.

Alzheimer's disease (AD) is a complex, multifactorial disorder, influenced by a multitude of variables ranging from genetic factors, age, and head injuries to vascular diseases, infections, and various other environmental and demographic determinants. Among the environmental factors, the role of the microbiome in the genesis of neurodegenerative disorders (NDs) is gaining increased recognition. This paradigm shift is substantiated by an extensive body of scientific literature, which underscores the significant contributions of microorganisms, encompassing viruses and gut-derived bacteria, to the pathogenesis of AD. The mechanism by which microbial infection exerts its influence on AD hinges primarily on inflammation. Neuroinflammation, activated in response to microbial infections, acts as a defense mechanism for the brain but can inadvertently lead to unexpected neuropathological perturbations, ultimately contributing to NDs. Given the ongoing uncertainty surrounding the genetic factors underpinning ND, comprehensive investigations into environmental factors, particularly the microbiome and viral agents, are imperative. Recent advances in neuroscientific research have unveiled the pivotal role of non-coding RNAs (ncRNAs) in orchestrating various pathways integral to neurodegenerative pathologies. While the upstream regulators governing the pathological manifestations of microorganisms remain elusive, an in-depth exploration of the nuanced role of ncRNAs holds promise for the development of prospective therapeutic interventions. This review aims to elucidate the pivotal role of ncRNAs as master modulators in the realm of neurodegenerative conditions, with a specific focus on Alzheimer's disease.

Determining host factors contributing to the reactivation of JC virus in kidney transplant recipients.

BACKGROUND AND AIMS: The John Cunningham virus (JCV) is the established etiological agent of the polyomavirus-associated nephropathy among renal transplant recipients. In the present study, we aimed to determine the probable predictive factors leading to JCV replication in renal transplant patients. MATERIAL AND METHODS: Urine and plasma samples were collected from a total of 120 consecutive renal-transplanted patients without preliminary screening from Jan 2018 to Mar 2019. After DNA extraction, the simultaneous detection and quantification of JCV and BK polyomavirus (BKV) were conducted using a Real-time quantitative PCR method. Moreover, statistical analyses were performed using the statistical software packages, SPSS version 21. RESULTS: The prevalence of JCV viruria and viremia among renal transplant recipients were 26 (21.67%) and 20 (16.67%), respectively. A significant association was observed between the JCV and two risk factors, diabetes mellitus (P = 0.002) and renal stones (P = 0.015). The prevalence of JCV viremia among recipients who were grafted near time to sampling was significantly higher (P = 0.02). There was a statistically significant coexistence between BK and JC viruses among our patients (P = 0.029). The frequency of JCV viruria in males was reported almost three times more than in females (P = 0.005). The JCV shedding in urine was significantly associated with the tropical steroids like prednisolone acetate, which have been the standard regimen (P = 0.039). Multivariable analysis revealed duration of post-transplantation (OR, 0.89; P = 0.038), diabetes mellitus (OR, 1.85; P = 0.034), and renal stone (OR 1.10; P = 0.04) as independent risk factors associated with JCV viremia post-renal transplantation. CONCLUSION: It seems that the discovery of potential risk factors, including immunological and non-immunological elements, may offer a possible preventive or therapeutic approach in the JCV disease episodes. The results of this study may also help clarify the probable clinical risk factors involving in progressive multifocal leukoencephalopathy development.

Gut bacteria, bacteriophages, and probiotics: Tripartite mutualism to quench the SARS-CoV2 storm.

Patients with SARS-CoV-2 infection, exhibit various clinical manifestations and severity including respiratory and enteric involvements. One of the main reasons for death among covid-19 patients is excessive immune responses directed toward cytokine storm with a low chance of recovery. Since the balanced gut microbiota could prepare health benefits by protecting against pathogens and regulating immune homeostasis, dysbiosis or disruption of gut microbiota could promote severe complications including autoimmune disorders; we surveyed the association between the imbalanced gut bacteria and the development of cytokine storm among COVID-19 patients, also the impact of probiotics and bacteriophages on the gut bacteria community to alleviate cytokine storm in COVID-19 patients. In present review, we will scrutinize the mechanism of immunological signaling pathways which may trigger a cytokine storm in SARS-CoV2 infections. Moreover, we are explaining in detail the possible immunological signaling pathway-directing by the gut bacterial community. Consequently, the specific manipulation of gut bacteria by using probiotics and bacteriophages for alleviation of the cytokine storm will be investigated. The tripartite mutualistic cooperation of gut bacteria, probiotics, and phages as a candidate prophylactic or therapeutic approach in SARS-CoV-2 cytokine storm episodes will be discussed at last.

The Effectiveness of the Anteroom (Vestibule) Area on Hospital Infection Control and Health Staff Safety: A Systematic Review.

The emergence of SARS-CoV2 in 2019 showed again that the world's healthcare system is not fully equipped and well-designed for preventing the transmission of nosocomial respiratory infections. One of the great tools for preventing the spread of infectious organisms in hospitals is the anteroom. Several articles have investigated the role of the anteroom in disease control but the lack of a comprehensive study in this field prompted us to provide more in-depth information to fill this gap. Also, this study aimed to assess the necessity to construct an anteroom area for hospital staff members at the entrance of each ward of the hospital, and specify the equipment and facilities which make the anteroom more efficient. Articles were identified through searches of Scopus, Web of Sciences, PubMed, and Embase for studies published in English until May 2020 reporting data on the effect of the anteroom (vestibule) area in controlling hospital infections. Data from eligible articles were extracted and presented according to PRISMA's evidence-based data evaluation search strategy. Also, details around the review aims and methods were registered with the PROSPERO. From the database, 209 articles were identified, of which 25 studies met the study criteria. Most studies demonstrated that an anteroom significantly enhances practical system efficiency. The results showed that the equipment such as ventilation system, high-efficiency particulate absorption filter, hand dispensers, alcohol-based disinfection, sink, mirror, transparent panel, UVC disinfection, and zone for PPE change, and parameters like temperature, door type, pressure, and size of the anteroom are factors that are effective on the safety of the hospital environment. Studies demonstrated that providing an anteroom for changing clothing and storing equipment may be useful in reducing the transmission of airborne infections in hospitals. Since the transmission route of SARS-CoV2 is common with other respiratory infectious agents, it can be concluded that a well-designed anteroom could potentially decrease the risk of SARS-CoV2 transmission during hospitalization as well.

Whole-transcriptome sequencing-based concomitant detection of viral and human genetic determinants of cutaneous lesions.

Severe viral infections of the skin can occur in patients with inborn errors of immunity (IEI). We report an all-in-one whole-transcriptome sequencing-based method by RNA-Seq on a single skin biopsy for concomitantly identifying the cutaneous virome and the underlying IEI. Skin biopsies were obtained from healthy and lesional skin from patients with cutaneous infections suspected to be of viral origin. RNA-Seq was utilized as the first-tier strategy for unbiased human genome-wide rare variant detection. Reads unaligned to the human genome were utilized for the exploration of 926 viruses in a viral genome catalog. In 9 families studied, the patients carried pathogenic variants in 6 human IEI genes, including IL2RG, WAS, CIB1, STK4, GATA2, and DOCK8. Gene expression profiling also confirmed pathogenicity of the human variants and permitted genome-wide homozygosity mapping, which assisted in identification of candidate genes in consanguineous families. This automated, online, all-in-one computational pipeline, called VirPy, enables simultaneous detection of the viral triggers and the human genetic variants underlying skin lesions in patients with suspected IEI and viral dermatosis.

High Seroprevalence of SARS-CoV-2 (COVID-19)-Specific Antibodies among Healthcare Workers: A Cross-Sectional Study in Guilan, Iran.

Background: This study was conducted to evaluate the anti-SARS-CoV-2 IgM and IgG antibodies among healthcare workers in Guilan. Methods: This cross-sectional study was conducted on 503 healthcare workers. Between April and May 2020, blood samples were collected from the healthcare workers of Razi Hospital in Rasht, Guilan, Iran. Enzyme-linked immunosorbent assay (ELISA) was used for the detection and quantitation of anti-SARS-CoV-2 IgM/IgG antibodies by using kits made by Pishtaz Teb Company, Tehran, Iran. Results: From a total of 503 participants, the result of the anti-SARS-CoV-2 IgM antibody test was positive in 28 subjects (5.6%) and the anti-SARS-CoV-2 IgG antibody test was positive in171 subjects (34%). Participants in the age group of 35-54 years were significantly more likely to have a positive anti-SARS-CoV-2 antibody test than the age group of 20-34 years (odds ratio = 1.53, 95% CI: 1.04-2.25, P=0.029). Also, physicians were significantly more likely to have a positive antibody test than office workers (odds ratio = 1.92, 95% CI: 1.04-3.54, P=0.037). The wide range of symptoms was significantly associated with the positive anti-SARS-CoV-2 antibody test. The most significant association was observed between fever and a positive anti-SARS-CoV-2 antibody test (odds ratio = 3.03, 95% CI: 2.06-4.44, P < 0.001). Conclusion: The results of the current study indicated that the seroprevalence of COVID-19 was high among healthcare workers of Guilan Province. It seems that this finding was due to the earlier exposure to COVID-19 and the lack of awareness and preparedness to deal with the pandemic in Iran, compared to other countries.

Toward a universal influenza virus vaccine: Some cytokines may fulfill the request.

The influenza virus annually causes widespread damages to the health and economy of the global community. Vaccination is currently the most crucial strategy in reducing the number of patients. Genetic variations, the high diversity of pandemic viruses, and zoonoses make it challenging to select suitable strains for annual vaccine production. If new pandemic viruses emerge, it will take a long time to produce a vaccine according to the new strains. In the present study, intending to develop a universal influenza vaccine, new bicistronic DNA vaccines were developed that expressed NP or NPm antigen with one of modified IL-18/ IL-17A/ IL-22 cytokine adjuvants. NPm is a mutant form of the antigen that has the ability for cytoplasmic accumulation. In order to investigate and differentiate the role of each of the components of Th1, Th2, Th17, and Treg cellular immune systems in the performance of vaccines, Treg competent and Treg suppressed mouse groups were used. Mice were vaccinated with Foxp3-FC immunogen to produce Treg suppressed mouse groups. The potential of the vaccines to stimulate the immune system was assessed by IFN-γ/IL-17A Dual FluoroSpot. The vaccine's ability to induce humoral immune response was determined by measuring IgG1, IgG2a, and IgA-specific antibodies against the antigen. Kinetics of Th1, Th2, and Th17 cellular immune responses after vaccination, were assessed by evaluating the expression changes of IL-17A, IFN-γ, IL-18, IL-22, IL-4, and IL-2 cytokines by semi-quantitative real-time RT-PCR. To assess the vaccines' ability to induce heterosubtypic immunity, challenge tests with homologous and heterologous viruses were performed and then the virus titer was measured in the lungs of animals. Evaluation of the data obtained from this study showed that the DNA-vaccines coding NPm have more ability to induces a potent cross-cellular immune response and protective immunity than DNA-vaccines coding NP. Although the use of IL-18/ IL-17A/ IL-22 genetic adjuvants enhanced immune responses and protective immunity, Administration of NPm in combination with modified IL-18 (Igk-mIL18-IgFC) induced the most effective immunity in Treg competent mice group.

Persistence of SARS-CoV-2 RNA in the nasopharyngeal, blood, urine, and stool samples of patients with COVID-19: a hospital-based longitudinal study.

BACKGROUND: The persistence of severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) RNA in the body fluids of patients with the novel coronavirus disease 2019 (COVID-19) may increase the potential risk of viral transmission. There is still uncertainty on whether the recommended quarantine duration is sufficient to reduce the risk of transmission. This study aimed to investigate the persistence of SARS-CoV-2 RNA in the nasopharyngeal, blood, urine, and stool samples of patients with COVID-19. METHODS: In this hospital-based longitudinal study, 100 confirmed cases of COVID-19 were recruited between March 2020 and August 2020 in Guilan Province, north of Iran. Nasopharyngeal, blood, urine, and stool samples were obtained from each participant at the time of hospital admission, upon discharge, 1 week after discharge, and every 2 weeks until all samples were negative for SARS-CoV-2 RNA by reverse transcription-polymerase chain reaction (RT-PCR) assay. A survival analysis was also performed to identify the duration of viral persistence. RESULTS: The median duration of viral RNA persistence in the nasopharyngeal samples was 8 days from the first positive RT-PCR result upon admission (95% CI 6.91-9.09); the maximum duration of viral shedding was 25 days from admission. Positive blood, urine, and stool RT-PCR results were detected in 24%, 7%, and 6% of the patients, respectively. The median duration of viral persistence in the blood, urine, and stool samples was 7 days (95% CI 6.07-7.93), 6 days (95% CI 4.16-8.41), and 13 days (95% CI 6.96-19.4), respectively. Also, the maximum duration of viral persistence in the blood, urine, and stool samples was 17, 11, and 42 days from admission, respectively. CONCLUSION: According to the present results, immediately after the hospitalized patients were discharged, no evidence of viral genetic materials was found. Therefore, appropriate treatments were selected for the patients at this hospital. However, we recommend further investigations on a larger sample size in multi-center and prospective randomized controlled trials (RCTs) to evaluate the effects of different drugs on the shedding of the virus through body secretions.

Functional and Structural Characterization of SARS-Cov-2 Spike Protein: An In Silico Study.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of the global outbreak of coronavirus disease 2019 (Covid-19), which has been considered as a pandemic by WHO. SARS-CoV-2 encodes four major structural proteins, among which spike protein has always been a main target for new vaccine studies. This in silico study aimed to investigate some physicochemical, functional, immunological, and structural features of spike protein using several bioinformatics tools. METHOD: We retrieved all SARS-CoV-2 spike protein sequences from different countries registered in NCBI GenBank. CLC Sequence Viewer was employed to translate and align the sequences, and several programs were utilized to predict B-cell epitopes. Modification sites such as phosphorylation, glycosylation, and disulfide bonds were defined. Secondary and tertiary structures of all sequences were further computed. RESULTS: Some mutations were determined, where only one (D614G) had a high prevalence. The mutations did not impact the B-cell and physicochemical properties of the spike protein. Seven disulfide bonds were specified and also predicted in several N-link glycosylation and phosphorylation sites. The results also indicated that spike protein is a non-allergen. CONCLUSION: In summary, our findings provided a deep understanding of spike protein, which can be valuable for future studies on SARS-CoV-2 infections and design of new vaccines.

The Prevalence, Etiological Agents, Clinical Features, Treatment, and Diagnosis of HIV-Associated Oral Candidiasis in Pediatrics Across the World: A Systematic Review and Meta-Analysis.

In HIV-infected pediatrics, oral candidiasis (OC) is a global issue of concern due to its association with dysphagia, malnutrition, and mortality. The present systematic review and meta-analysis are the first to determine the prevalence of OC in HIV-infected pediatrics worldwide. We searched international (PubMed, Web of Science, Scopus, and Embase) databases for studies published between January 2000 to May 2020 reporting the epidemiologic features of OC in HIV-infected pediatrics. Inclusion and exclusion criteria were defined to select eligible studies. Data were extracted and presented according to PRISMA guidelines. The results of the meta-analysis were visualized as a forest plot. Heterogeneity was also analyzed using the I 2, and τ2 statistics. The publication bias was evaluated using Egger test. The literature search revealed 1926 studies, of which 34 studies met the eligibility criteria, consisting of 4,474 HIV-infected pediatrics from 12 different countries. The overall prevalence of OC among HIV-infected pediatrics was 23.9% (95% CI 17.3-32.0%), and Candida albicans was the most prevalent etiologic agent. Pseudomembranous candidiasis was the predominant clinical manifestation in HIV-infected pediatrics suffering from OC. Thirty articles involving 4,051 individuals provided data on HIV treatment status. Among the 4,051 individuals, 468 (11.53%) did not receive HIV treatment. The data from 11 articles demonstrated that HIV treatment was significantly associated with a reduction in oral Candida colonization or infection. In contrast, others showed the opposite relationship or did not report any statistical data. A high level of I 2 (I 2 = 96%, P < 0.01) and τ2 (τ2 = 1.36, P < 0.01) was obtained among studies, which provides evidence of notable heterogeneity between studies. OC is approximately frequent in HIV-positive children. Therefore, efforts should be made to teach dental and non-dental clinicians who care for HIV-infected pediatrics to diagnose and treat this infection.

Analysis of risk factors influencing the BK polyomavirus replication in patients with ESRD waiting for kidney transplantation.

BACKGROUND: Since the pre-transplant status affects the renal transplantation success and ultimately the survival rate, identifying the probable risk factors that increase the chance of BK virus replication in end-stage renal disease patients can be included in proposing proper surveillance guidelines during pre and post-transplantation. METHODS: A descriptive cross-sectional study was performed by collecting plasma samples from 192 ESRD patients undergoing hemodialysis for at least 3 months. Quantitative Real-time PCR assay was used to detect and measure the BK viral load. Demographic and clinical characteristics of the patients who had BK viremia were documented. RESULTS: 14 (7.3%) out of our 192 participants had BK virus viremia (95%CI 4.2%-11.6%). Demographic characteristics including etiology of ESRD and underlying diseases, mean duration and frequency of dialysis, co-infection with HBV and HCV did not affect the virus replication, since the difference between patients with BK virus viremia and BK virus negative individuals was not statistically significant. However, the statistical significance of the mean age of men with BKV and without BK virus viremia was found (OR: 3.42, P = 0.02 95%CI 0.86-13.61). Also, multiple regression analyses of some other parameters revealed that old age, high body mass index and male gender can be predictive factors of BK virus viremia in ESRD patients. CONCLUSION: Based on our findings, elderly male had higher chance of being exposed to BK virus viremia. Some other demographic characteristics such as a high BMI, old age and gender (male) can increase the risk of BK viremia in patients with ESRD prior to kidney transplantation.

The Phylogenetic Relatedness of bla NDM-1 Harboring Extended-Spectrum ß-Lactamase Producing Uropathogenic Escherichia coli and Klebsiella pneumoniae in the North of Iran.

BACKGROUND: Escherichia coli and Klebsiella pneumoniae as an important part of Enterobacterales family are important causes of both community- and hospital-acquired infections. The present study aimed to investigate the prevalence of antibiotics resistance and molecular characteristics of uropathogenic isolates of E. coli and K. pneumoniae in Iranian patients. METHODS: This cross-sectional study performed on 223 Escherichia coli and 68 Klebsiella pneumoniae isolates obtained from hospitalized patients in the north of Iran. The isolates were identified by standard microbiologic tests and confirmed by API 20E strip. Disk diffusion method was applied to determine antibiotic susceptibility pattern. The presence of ß-lactamases encoding genes was evaluated by PCR method. Analysis of the mutations and homology among sequences was done by the CLC sequence viewer (Qiagen, Denmark), and phylogenetic trees were constructed by the neighbor-joining method (Bootstrap: 1000 times). RESULTS: The overall rates of extended-spectrum ß-lactamases (ESBLs)-producing E. coli and K. pneumoniae isolates were 37.7% and 32.4%, respectively. The overall presence of bla SHV, bla NDM-1, and bla OXA-1 genes was detected in 16 (5.5%), 12 (4.1%), and 48 (16.4%) of isolates, respectively. The neighbor-joining analysis for E. coli KU985246.1 strain showed that the most related bla NDM-1 sequences were from China, Singapore, UK, Thailand, and Bangladesh. While K. pneumoniae KU985245.1 strains were mostly related to bla NDM-1 sequences form Myanmar, and China. CONCLUSION: In summary, the remarkable rate of ESBL-producing uropathogenic Enterobacterales along with the first prevalence of NDM-1 ß-lactamases can be a serious concern in our region.

In vitro activity of colistin against multidrug-resistant Acinetobacter baumannii isolates harboring blaOXA-23-like and blaOXA-24-like genes: A multicenter based study.

This study was aimed to evaluate occurrence of antibiotic resistance and the presence of resistance determinants among clinical isolates of Acinetobacter baumannii. This cross-sectional study from January to September 2018 was performed on 59 A. baumannii strains isolated from clinical samples in the north of Iran. Isolates were identified by standard microbiologic tests and molecular method. Antimicrobial susceptibility testing was carried out by disk diffusion and broth microdilution methods. The presence of carbapenem resistance genes was detected by PCR method. All isolates were resistant to cefepime, meropenem, imipenem and ceftazidime. The lowest resistance rate was observed against doxycycline with 33.9%. Minimum inhibitory concentration (MIC) results showed that all carbapenem-resistant A. baumannii (CRAB) isolates were susceptible to colistin with MIC50 and MIC90 values of 1/2 µg/mL. Among 59 CRAB, blaOXA-23-like was the most prevalent gene (86.4%) followed by blaOXA-24-like (69.5%). Meanwhile, none of the clinical isolates harbored blaOXA-58-like gene. We found a high prevalence of CRAB strains harboring OXA-type carbapenemases in the north of Iran. Our results suggests that the presence of OXA-type genes was not directly correlated with the increase of imipenem MIC level, but can be clinically important as they contribute to the selection of CRAB strains.

Co-expression of Interleukin-17A molecular adjuvant and prophylactic Helicobacter pylori genetic vaccine could cause sterile immunity in Treg suppressed mice.

The increasing clinical significance of Helicobacter pylori (H. pylori) in human stomach cancer has led to global efforts to eradicate this pathogen. Recent studies have confirmed the importance of some cytokines such as Interleukin-18 (IL-18), Interleukin-8 (IL-8), Interleukin-17A (IL-17A) and Interleukin-22 (IL-22) in the pathogenesis of the so-called bacterium. This study was designed to compare the effects of Type 1T helper (Th1), Type 2T helper (Th2) cells, Regulatory T cells (Treg) and T helper 17 (Th17) modulatory effects on the efficacy of designed H. pylori vaccine by incorporating some molecular adjuvants in Treg competent and Treg suppressed groups. A bicistronic vector was used for simultaneous expression of codon-optimized Outer inflammatory protein a (OipA) gene and modified mice IL-18, IL-17A, IL-22 and Foxp3 (forkhead box P3) cytokines from four cassettes. Immunization of mice groups was performed using produced plasmids intradermally. Specific IgG1 and IgG2 and IgA antibody titers produced in mice were confirmed by enzyme-linked immunosorbent assay (ELISA) in sera and intestine obtained four weeks after the last immunization. After being stimulated with a mixture of both anti-CD28 mAb and H. pylori lysate, frequencies of single Interferon-Gamma (IFN-γ), single IL-17 and dual IFN-γ/IL-17-secreting T-cells were documented using dual-color FluoroSpot. The kinetics of Th1, Th2 and Th17 in the immunized animals was determined by relative quantification of IL-17A, IL-22, IFN-γ, IL-8, IL-2 and IL-4 specific mRNAs. Four weeks after bacterial challenge, quantitative colony count in the isolated and homogenized stomachs was utilized to assess the level of protective immunity among all groups. The results of immunologic assays showed that the highest cell-mediated immunity cytokines were produced in IL-17 receiving group in which the Treg responses were suppressed previously by the administration of the Foxp3 as an immunogen. In addition, potent clearance of Helicobacter pylori infection was seen in this group as well.

Analysis of babA, cagE and cagA Genes in Helicobacter pylori from Upper Gastric Patients in the North of Iran.

OBJECTIVE: Helicobacter pylori is a Gram-negative bacterium which has a serious effect on up to half of the world's population and has been related to different gastric diseases. The goal of this study was to assess the frequency of babA, cagE and cagA genotypes among H. pylori strains isolated from gastric biopsies of endoscopic patients in the north of Iran. METHODS: The present study was performed on 90 strains of H. pylori isolated from patients with gastric diseases (Gastric ulcer (GU), Duodenal ulcer (DU), Gastritis (G), Non-ulcer dyspepsia (NUD) and Gastric adenocarcinoma (GC)). DNA was extracted from all isolated strains and PCR method was performed to detect the prevalence of babA2, cagE and cagA genes using specific primers. RESULTS: Among 90 samples of H. pylori, babA2, cagE, and cagA genes were detected in 42.2%, 30% and 82.2% of strains respectively. The statistical analysis showed that the prevalence of cagA gene in GU, G, DU, and NUD was significantly higher than other genes. Moreover, cagA, and babA2 genes were significantly more prevalent in GC patients compared to cagE gene. Our isolates exhibited 8 distinct arrangements of virulence patterns. The occurrence of cagA (35.6%) was the most prevalent pattern followed by cagA/babA2 (20%) and cagA/babA2/cagE (14.4%). CONCLUSION: In summary, as first report from Guilan province in the north of Iran, we showed significant association between the presence of babA2, cagE, and cagA genes in different types of gastric disorders.