RESUMEN

Antigen-specific cytotoxic CD8 T cells are extremely effective in controlling tumor growth and have been the focus of immunotherapy approaches. We leverage in silico tools to investigate whether the occurrence of mutations in proteins previously described as immunogenic and highly expressed by glioblastoma multiforme (GBM), such as Epidermal Growth Factor Receptor (EGFR), Isocitrate Dehydrogenase 1 (IDH1), Phosphatase and Tensin homolog (PTEN) and Tumor Protein 53 (TP53), may be contributing to the differential presentation of immunogenic epitopes. We recovered Class I MHC binding information from wild-type and mutated proteins using the Immune Epitope Database (IEDB). After that, we built peptide-MHC (pMHC-I) models in HLA-arena, followed by hierarchical clustering analysis based on electrostatic surface features from each complex. We identified point mutations that are determinants for the presentation of a set of peptides from TP53 protein. We point to structural features in the pMHC-I complexes of wild-type and mutated peptides, which may play a role in the recognition of CD8 T cells. To further explore these features, we performed 100 ns molecular dynamics simulations for the peptide pairs (wt/mut) selected. In pursuit of novel therapeutic targets for GBM treatment, we selected peptides where our predictive results indicated that mutations would not disrupt epitope presentation, thereby maintaining a specific CD8 T cell immune response. These peptides hold potential for future GBM interventions, including peptide-based or mRNA vaccine development applications.

Asunto(s)

Presentación de Antígeno , Linfocitos T CD8-positivos , Glioblastoma , Isocitrato Deshidrogenasa , Proteína p53 Supresora de Tumor , Glioblastoma/inmunología , Glioblastoma/genética , Glioblastoma/terapia , Humanos , Linfocitos T CD8-positivos/inmunología , Isocitrato Deshidrogenasa/genética , Isocitrato Deshidrogenasa/inmunología , Isocitrato Deshidrogenasa/química , Proteína p53 Supresora de Tumor/genética , Proteína p53 Supresora de Tumor/inmunología , Presentación de Antígeno/inmunología , Mutación , Fosfohidrolasa PTEN/genética , Fosfohidrolasa PTEN/inmunología , Fosfohidrolasa PTEN/química , Receptores ErbB/inmunología , Receptores ErbB/genética , Epítopos de Linfocito T/inmunología , Epítopos de Linfocito T/genética , Antígenos de Histocompatibilidad Clase I/inmunología , Antígenos de Histocompatibilidad Clase I/genética , Neoplasias Encefálicas/inmunología , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/terapia

RESUMEN

BACKGROUND: Bariatric surgery is an effective intervention that causes a series of metabolic changes related to inflammatory processes; however, the variation of biomarkers related to these processes is not entirely understood. Our objective was to investigate the variation of modulation and expression of biomarkers associated with inflammation in patients who underwent bariatric surgery. METHODS: We searched the MEDLINE (via PubMed), EMBASE (via Elsevier), Cochrane Central Register of Controlled Trials, Latin American and Caribbean Literature on Health Sciences (via virtual health library), Cumulative Index to Nursing and Allied Health Literature (via EBSCO), Web of Science core collection, and Scopus (via Elsevier) databases, and the gray literature was examined from inception to January 2022. Three pairs of reviewers performed data screening, extraction, and quality assessment independently. Meta-analysis with random effects models was used for general, subgroup, and sensitivity analyses. The I2 statistic was used to assess heterogeneity between studies. RESULTS: In total, 96 articles were included in this systematic review; of these, 87 studies met the criteria for the meta-analysis, involving 3,533 participants. Five biomarkers were included in the meta-analysis (tumor necrosis factor alpha; interleukin 6; leptin; interleukin 1 beta, and lipopolysaccharides). Only leptin showed a significant decrease in the first month after surgery (mean difference -20.71; [95% confidence interval: -28.10 to -13.32, P < .0001; I2 = 66.7%), with moderate heterogeneity. The 12 months after surgery showed a significant decrease in tumor necrosis factor alpha (mean difference -0.89; [95% confidence interval: -1.37 to -0.42], P = .0002; I2 = 94.7%), interleukin 6 (mean difference -1.62; [95% confidence interval: -1.95 to -1.29], P < .0001; I2 = 94.9%), leptin (mean difference -28.63; [95% confidence interval: -34.02 to -23.25], P < .0001; I2 = 92.7%), and interleukin 1 beta (mean difference -2.46; [95% confidence interval: -4.23 to -0.68], P = .006; I2 = 98.3%), all with high heterogeneity. The type of surgery did not show significant differences for the biomarkers at the first month and 12 months, and the results have not changed with high-quality studies. In the 12-month measurement, variations in tumor necrosis factor alpha and leptin were associated with body mass index. CONCLUSION: The findings of this meta-analysis suggest that Roux-en-Y gastric bypass and sleeve gastrectomy bariatric surgeries are associated with a significant reduction in leptin at 1 month after bariatric surgical intervention and tumor necrosis factor alpha, leptin, and interleukin 1 beta after 12 months.

RESUMEN

BACKGROUND: Gut microbiota-derived short-chain fatty-acid (SFCA) acetate protects mice against RSV A2 strain infection by increasing interferon-ß production and expression of interferon-stimulated genes (ISGs). However, the role of SFCA in RSV infection using strains isolated from patients is unknown. METHODS: We first used RSV clinical strains isolated from infants hospitalized with RSV bronchiolitis to investigate the effects of in vitro SCFA-acetate treatment of human pulmonary epithelial cells. We next examined whether SCFA-acetate treatment is beneficial in a mouse model of RSV infection using clinical isolates. We sought to investigate the relationship of gut microbiota and fecal acetate with disease severity among infants hospitalized with RSV bronchiolitis, and whether treating their respiratory epithelial cells with SCFA-acetate ex-vivo impacts viral load and ISG expression. We further treated epithelial cells from SARS-CoV-2 infected patients with SCFA-acetate. FINDINGS: In vitro pre-treatment of A549 cells with SCFA-acetate reduced RSV infection with clinical isolates and increased the expression of RIG-I and ISG15. Animals treated with SCFA-acetate intranasally recovered significantly faster, with reduction in the RSV clinical isolates viral load, and increased lung expression of IFNB1 and the RIG-I. Experiments in RIG-I knockout A549 cells demonstrated that the protection relies on RIG-I presence. Gut microbial profile was associated with bronchiolitis severity and with acetate in stool. Increased SCFA-acetate levels were associated with increasing oxygen saturation at admission, and shorter duration of fever. Ex-vivo treatment of patients' respiratory cells with SCFA-acetate reduced RSV load and increased expression of ISGs OAS1 and ISG15, and virus recognition receptors MAVS and RIG-I, but not IFNB1. These SCFA-acetate effects were not found on cells from SARS-CoV-2 infected patients. INTERPRETATION: SCFA-acetate reduces the severity of RSV infection and RSV viral load through modulation of RIG-I expression. FUNDING: FAPERGS (FAPERGS/MS/CNPq/SESRS no. 03/2017 - PPSUS 17/2551-0001380-8 and COVID-19 20/2551-0000258-6); CNPq 312504/2017-9; CAPES) - Finance Code 001.

Asunto(s)

Bronquiolitis , COVID-19 , Infecciones por Virus Sincitial Respiratorio , Virus Sincitial Respiratorio Humano , Acetatos/metabolismo , Acetatos/farmacología , Animales , Antivirales/metabolismo , Antivirales/farmacología , Antivirales/uso terapéutico , Bronquiolitis/tratamiento farmacológico , Bronquiolitis/metabolismo , Ácidos Grasos Volátiles/metabolismo , Humanos , Lactante , Pulmón/metabolismo , Ratones , Infecciones por Virus Sincitial Respiratorio/tratamiento farmacológico , Infecciones por Virus Sincitial Respiratorio/genética , Virus Sincitial Respiratorio Humano/fisiología , SARS-CoV-2

RESUMEN

Acute bronchiolitis caused by the respiratory syncytial virus triggers an inflammatory response with the production and release of several pro-inflammatory cytokines. Evidence suggests that their levels are associated with the severity of the infection. This systematic review and meta-analysis aim to assess whether the levels of TNF-α and IFN-γ are associated with the severity of acute viral bronchiolitis. We searched MEDLINE libraries (via PUBMED), EMBASE, Cochrane Central Register of Controlled Trials (CENTRAL), Scientific Electronic Library Online (SciELO), Latin American Caribbean Health Sciences Literature (LILACS), Cumulative Index to Nursing and Allied Health Literature (CINAHL), Web of Science, and the gray literature through April 2020. Random effect models were used for general and subgroup analysis. In total, six studies were included with a total of 744 participants. The mean TNF-α levels between the severe group did not differ from the control group 0.14 (95% CI: -0.53 to 0.82, I2 = 91%, p < 0.01); the heterogeneity was high. The results remained insignificant when the analyses were performed including only studies with high quality 0.25 (95% CI: -0.46 to 0.96, I2 = 92%, p < 0.01) I2 = 95%, p = 0.815), when TNF-α was nasal 0.60 (95% CI: -0.49 to 1.69), I2 = 94%, p < 0.01), or serum -0.08 (95% CI: -0.48 to 0.31), I2 = 29%, p = 0.24). In the analysis of studies measuring IFN-γ, there was also no significance of -0.67 (95% CI: -1.56 to 0.22, I2 = 76%, p = 0.04). In conclusion, this meta-analysis suggests that the most severe patients do not have different mean TNF-α and IFN-γ values ​than patients with mild disease, but the heterogeneity of the studies was high. Supplemental data for this article is available online at https://doi.org/10.1080/08830185.2021.1889534.

Asunto(s)

Bronquiolitis Viral , Bronquiolitis , Citocinas , Humanos , Factor de Necrosis Tumoral alfa

RESUMEN

Although not being the first viral pandemic to affect humankind, we are now for the first time faced with a pandemic caused by a coronavirus. The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has been responsible for the COVID-19 pandemic, which caused more than 4.5 million deaths worldwide. Despite unprecedented efforts, with vaccines being developed in a record time, SARS-CoV-2 continues to spread worldwide with new variants arising in different countries. Such persistent spread is in part enabled by public resistance to vaccination in some countries, and limited access to vaccines in other countries. The limited vaccination coverage, the continued risk for resistant variants, and the existence of natural reservoirs for coronaviruses, highlight the importance of developing additional therapeutic strategies against SARS-CoV-2 and other coronaviruses. At the beginning of the pandemic it was suggested that countries with Bacillus Calmette-Guérin (BCG) vaccination programs could be associated with a reduced number and/or severity of COVID-19 cases. Preliminary studies have provided evidence for this relationship and further investigation is being conducted in ongoing clinical trials. The protection against SARS-CoV-2 induced by BCG vaccination may be mediated by cross-reactive T cell lymphocytes, which recognize peptides displayed by class I Human Leukocyte Antigens (HLA-I) on the surface of infected cells. In order to identify potential targets of T cell cross-reactivity, we implemented an in silico strategy combining sequence-based and structure-based methods to screen over 13,5 million possible cross-reactive peptide pairs from BCG and SARS-CoV-2. Our study produced (i) a list of immunogenic BCG-derived peptides that may prime T cell cross-reactivity against SARS-CoV-2, (ii) a large dataset of modeled peptide-HLA structures for the screened targets, and (iii) new computational methods for structure-based screenings that can be used by others in future studies. Our study expands the list of BCG peptides potentially involved in T cell cross-reactivity with SARS-CoV-2-derived peptides, and identifies multiple high-density "neighborhoods" of cross-reactive peptides which could be driving heterologous immunity induced by BCG vaccination, therefore providing insights for future vaccine development efforts.

Asunto(s)

Vacuna BCG/inmunología , COVID-19/inmunología , Reacciones Cruzadas/inmunología , Péptidos/inmunología , SARS-CoV-2/inmunología , Linfocitos T/inmunología , Vacunas Virales/inmunología , Humanos , Pandemias/prevención & control , Vacunación/métodos

RESUMEN

HLA-G is considered to be an immune checkpoint molecule, a function that is closely linked to the structure and dynamics of the different HLA-G isoforms. Unfortunately, little is known about the structure and dynamics of these isoforms. For instance, there are only seven crystal structures of HLA-G molecules, being all related to a single isoform, and in some cases lacking important residues associated to the interaction with leukocyte receptors. In addition, they lack information on the dynamics of both membrane-bound HLA-G forms, and soluble forms. We took advantage of in silico strategies to disclose the dynamic behavior of selected HLA-G forms, including the membrane-bound HLA-G1 molecule, soluble HLA-G1 dimer, and HLA-G5 isoform. Both the membrane-bound HLA-G1 molecule and the soluble HLA-G1 dimer were quite stable. Residues involved in the interaction with ILT2 and ILT4 receptors (α3 domain) were very close to the lipid bilayer in the complete HLA-G1 molecule, which might limit accessibility. On the other hand, these residues can be completely exposed in the soluble HLA-G1 dimer, due to the free rotation of the disulfide bridge (Cys42/Cys42). In fact, we speculate that this free rotation of each protomer (i.e., the chains composing the dimer) could enable alternative binding modes for ILT2/ILT4 receptors, which in turn could be associated with greater affinity of the soluble HLA-G1 dimer. Structural analysis of the HLA-G5 isoform demonstrated higher stability for the complex containing the peptide and coupled ß2-microglobulin, while structures lacking such domains were significantly unstable. This study reports for the first time structural conformations for the HLA-G5 isoform and the dynamic behavior of HLA-G1 molecules under simulated biological conditions. All modeled structures were made available through GitHub (https://github.com/KavrakiLab/), enabling their use as templates for modeling other alleles and isoforms, as well as for other computational analyses to investigate key molecular interactions.

Asunto(s)

Membrana Celular/metabolismo , Antígenos HLA-G/metabolismo , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Antígenos HLA-G/química , Antígenos HLA-G/genética , Humanos , Membrana Dobles de Lípidos , Dominios y Motivos de Interacción de Proteínas , Isoformas de Proteínas , Multimerización de Proteína , Estabilidad Proteica , Relación Estructura-Actividad

RESUMEN

Cystic fibrosis (CF) is a genetic disease caused by variants in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. There are over 2,000 different pathogenic and non-pathogenic variants described in association with a broad clinical heterogeneity. In this work, we identified a novel variant S511Lfs*2 in CFTR gene that has not been reported in patients with CF. The patient was a female genotyped with c.1000C>T (legacy name: R334W) variant (pathogenic, CF-causing) and the novel variant (S511Lfs*2). We verified the amino acid sequence, the protein structure, and predicted the pathogenicity employing computational analysis. Our findings showed that S511Lfs*2 is a frameshift variant and suggest that it is associated with severe CF phenotype, as it leads to a lack of CFTR protein synthesis, and consequently the loss of its functional activity.

Asunto(s)

Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Fibrosis Quística/genética , Mutación del Sistema de Lectura , Adulto , Femenino , Humanos , Fenotipo , Adulto Joven

RESUMEN

The use of model organisms for recombinant protein production results in the addition of model-specific post-translational modifications (PTMs) that can affect the structure, charge, and function of the protein. The 70-kDa heat shock proteins (Hsp70) were originally described as intracellular chaperones, with ATPase and foldase activity. More recently, new extracellular activities of Hsp70 proteins (e.g. as immunomodulators) have been identified. While some studies indicate an inflammatory potential for extracellular Hsp70 proteins, others suggest an immunosuppressive activity. We hypothesized that the production of recombinant Hsp70 in different expression systems would result in the addition of different PTMs, perhaps explaining at least some of these opposing immunological outcomes. We produced and purified Mycobacterium tuberculosis DnaK from two different systems, Escherichia coli and Pichia pastoris, and analyzed by mass spectrometry the protein preparations, investigating the impact of PTMs in an in silico and in vitro perspective. The comparisons of DnaK structures in silico highlighted that electrostatic and topographical differences exist that are dependent upon the expression system. Production of DnaK in the eukaryotic system dramatically affected its ATPase activity, and significantly altered its ability to downregulate MHC II and CD86 expression on murine dendritic cells (DCs). Phosphatase treatment of DnaK indicated that some of these differences related specifically to phosphorylation. Altogether, our data indicate that PTMs are an important characteristic of the expression system, with differences that impact interactions of Hsps with their ligands and subsequent functional activities.

RESUMEN

Respiratory syncytial virus (RSV) is a common cause of childhood lower respiratory tract infections. The recent failure of a vaccine candidate based on recombinant F protein underlines the urgent need to better understand the protective human memory immune response against RSV. Signal transducer and activator of transcription 3 (STAT3) protein is a transcription factor that promotes the maturation of the memory CD8 T cell response in cooperation with IL-10 and IL-21. However, the role of STAT3 in the memory CD8 T cell response during RSV infection remains to be elucidated. We found that in infants with bronchiolitis infected with RSV, the expression of STAT3 detected in nasal washes is reduced when compared to that in infants infected by other viruses. In vitro, RSV impairs STAT3 phosphorylation induced by IL-21 in purified human memory CD8 T cells. In addition, RSV decreases granzyme B production by memory CD8 T cells, reducing its cytotoxic activity against RSV-infected epithelial pulmonary cell lines. Together, these data indicate that RSV modulates the IL-21/STAT3 pathway in human memory CD8 T cells, and this could be a mechanism to be further explored to improve the memory response against the infection.

Asunto(s)

Linfocitos T CD8-positivos/inmunología , Interleucinas/inmunología , Infecciones por Virus Sincitial Respiratorio/inmunología , Virus Sincitial Respiratorio Humano/inmunología , Factor de Transcripción STAT3/inmunología , Linfocitos T CD8-positivos/patología , Linfocitos T CD8-positivos/virología , Células Cultivadas , Femenino , Interacciones Huésped-Patógeno , Humanos , Memoria Inmunológica , Lactante , Masculino , Modelos Moleculares , Fosforilación , Infecciones por Virus Sincitial Respiratorio/patología , Infecciones por Virus Sincitial Respiratorio/virología , Virus Sincitial Respiratorio Humano/fisiología

RESUMEN

Immunotherapy has become one of the most promising avenues for cancer treatment, making use of the patient's own immune system to eliminate cancer cells. Clinical trials with T-cell-based immunotherapies have shown dramatic tumor regressions, being effective in multiple cancer types and for many different patients. Unfortunately, this progress was tempered by reports of serious (even fatal) side effects. Such therapies rely on the use of cytotoxic T-cell lymphocytes, an essential part of the adaptive immune system. Cytotoxic T-cells are regularly involved in surveillance and are capable of both eliminating diseased cells and generating protective immunological memory. The specificity of a given T-cell is determined through the structural interaction between the T-cell receptor (TCR) and a peptide-loaded major histocompatibility complex (MHC); i.e., an intracellular peptide-ligand displayed at the cell surface by an MHC molecule. However, a given TCR can recognize different peptide-MHC (pMHC) complexes, which can sometimes trigger an unwanted response that is referred to as T-cell cross-reactivity. This has become a major safety issue in TCR-based immunotherapies, following reports of melanoma-specific T-cells causing cytotoxic damage to healthy tissues (e.g., heart and nervous system). T-cell cross-reactivity has been extensively studied in the context of viral immunology and tissue transplantation. Growing evidence suggests that it is largely driven by structural similarities of seemingly unrelated pMHC complexes. Here, we review recent reports about the existence of pMHC "hot-spots" for cross-reactivity and propose the existence of a TCR interaction profile (i.e., a refinement of a more general TCR footprint in which some amino acid residues are more important than others in triggering T-cell cross-reactivity). We also make use of available structural data and pMHC models to interpret previously reported cross-reactivity patterns among virus-derived peptides. Our study provides further evidence that structural analyses of pMHC complexes can be used to assess the intrinsic likelihood of cross-reactivity among peptide-targets. Furthermore, we hypothesize that some apparent inconsistencies in reported cross-reactivities, such as a preferential directionality, might also be driven by particular structural features of the targeted pMHC complex. Finally, we explain why TCR-based immunotherapy provides a special context in which meaningful T-cell cross-reactivity predictions can be made.

RESUMEN

Drug-induced liver injury (DILI) is a major cause of acute liver failure (ALF), where hepatocyte necrotic products trigger liver inflammation, release of CXC chemokine receptor 2 (CXCR2) ligands (IL-8) and other neutrophil chemotactic molecules. Liver infiltration by neutrophils is a major cause of the life-threatening tissue damage that ensues. A GRPR (gastrin-releasing peptide receptor) antagonist impairs IL-8-induced neutrophil chemotaxis in vitro. We investigated its potential to reduce acetaminophen-induced ALF, neutrophil migration, and mechanisms underlying this phenomenon. We found that acetaminophen-overdosed mice treated with GRPR antagonist had reduced DILI and neutrophil infiltration in the liver. Intravital imaging and cell tracking analysis revealed reduced neutrophil mobility within the liver. Surprisingly, GRPR antagonist inhibited CXCL2-induced migration in vivo, decreasing neutrophil activation through CD11b and CD62L modulation. Additionally, this compound decreased CXCL8-driven neutrophil chemotaxis in vitro independently of CXCR2 internalization, induced activation of MAPKs (p38 and ERK1/2) and downregulation of neutrophil adhesion molecules CD11b and CD66b. In silico analysis revealed direct binding of GRPR antagonist and CXCL8 to the same binding spot in CXCR2. These findings indicate a new potential use for GRPR antagonist for treatment of DILI through a mechanism involving adhesion molecule modulation and possible direct binding to CXCR2.

Asunto(s)

Bombesina/análogos & derivados , Enfermedad Hepática Inducida por Sustancias y Drogas/tratamiento farmacológico , Neutrófilos/inmunología , Fragmentos de Péptidos/farmacología , Receptores de Bombesina/antagonistas & inhibidores , Receptores de Interleucina-8B/metabolismo , Animales , Bombesina/farmacología , Movimiento Celular/efectos de los fármacos , Células Cultivadas , Enfermedad Hepática Inducida por Sustancias y Drogas/inmunología , Quimiotaxis/efectos de los fármacos , Humanos , Interleucina-8/metabolismo , Ratones , Ratones Endogámicos , Activación Neutrófila/efectos de los fármacos , Unión Proteica , Transducción de Señal/efectos de los fármacos

RESUMEN

The immune system is constantly challenged, being required to protect the organism against a wide variety of infectious pathogens and, at the same time, to avoid autoimmune disorders. One of the most important molecules involved in these events is the Major Histocompatibility Complex class I (MHC-I), responsible for binding and presenting small peptides from the intracellular environment to CD8(+) T cells. The study of peptide:MHC-I (pMHC-I) molecules at a structural level is crucial to understand the molecular mechanisms underlying immunologic responses. Unfortunately, there are few pMHC-I structures in the Protein Data Bank (PDB) (especially considering the total number of complexes that could be formed combining different peptides), and pMHC-I modelling tools are scarce. Here, we present DockTope, a free and reliable web-based tool for pMHC-I modelling, based on crystal structures from the PDB. DockTope is fully automated and allows any researcher to construct a pMHC-I complex in an efficient way. We have reproduced a dataset of 135 non-redundant pMHC-I structures from the PDB (Cα RMSD below 1 Å). Modelling of pMHC-I complexes is remarkably important, contributing to the knowledge of important events such as cross-reactivity, autoimmunity, cancer therapy, transplantation and rational vaccine design.

Asunto(s)

Linfocitos T CD8-positivos/metabolismo , Biología Computacional/métodos , Antígenos de Histocompatibilidad Clase I/metabolismo , Internet , Péptidos/metabolismo , Algoritmos , Secuencia de Aminoácidos , Bases de Datos de Proteínas , Epítopos/química , Epítopos/genética , Epítopos/metabolismo , Antígenos de Histocompatibilidad Clase I/química , Antígenos de Histocompatibilidad Clase I/genética , Humanos , Modelos Moleculares , Péptidos/química , Unión Proteica , Dominios Proteicos , Reproducibilidad de los Resultados

RESUMEN

The CrossTope is a highly curate repository of three-dimensional structures of peptide:major histocompatibility complex (MHC) class I complexes (pMHC-I). The complexes hosted by this databank were obtained in protein databases and by large-scale in silico construction of pMHC-I structures, using a new approach developed by our group. At this moment, the database contains 182 'non-redundant' pMHC-I complexes from two human and two murine alleles. A web server provides interface for database query. The user can download (i) structure coordinate files and (ii) topological and charges distribution maps images from the T-cell receptor-interacting surface of pMHC-I complexes. The retrieved structures and maps can be used to cluster similar epitopes in cross-reactivity approaches, to analyse viral escape mutations in a structural level or even to improve the immunogenicity of tumour antigens. Database URL: http://www.crosstope.com.br.

Asunto(s)

Bases de Datos de Proteínas , Complejo Mayor de Histocompatibilidad/inmunología , Modelos Moleculares , Péptidos/química , Péptidos/inmunología , Animales , Reacciones Cruzadas/inmunología , Cristalografía por Rayos X , Humanos , Almacenamiento y Recuperación de la Información , Ratones , Análisis Multivariante , Diseño de Software , Interfaz Usuario-Computador

RESUMEN

The Bunyaviridae virus family is composed by five genera, of which the Hantavirus genus is one of the most important representatives. Occasionally, these viruses can be transmitted to humans, giving rise to severe diseases that present high mortality rates. We analyzed the amino acid sequences of the nucleocapsid (N) proteins of 34 different hantaviruses to investigate the potential mechanisms involved in immunogenicity against hantaviruses. Immunogenic epitopes described in the literature through experimental analyses for Sin Nombre (SNV), Puumala (PUUV), and Hantaan (HTNV) viruses' species were retrieved. We identified and characterized the regions believed to be responsible for the induction of immune response in hosts. We found that N protein epitopes described in the literature for PUUV, SNV and HTNV viruses are all located in highly conserved regions of the protein. The high conservation of these regions suggests that a cross-reactive immune response among different hantaviruses can be induced.

Asunto(s)

Proteínas de la Cápside/inmunología , Secuencia Conservada , Epítopos/inmunología , Orthohantavirus/inmunología , Proteínas del Núcleo Viral/inmunología , Proteínas de la Cápside/química , Epítopos/química , Humanos , Homología de Secuencia de Aminoácido , Proteínas del Núcleo Viral/química

RESUMEN

Cellular immune response plays a central role in outcome of Hepatitis C Virus (HCV) infection. While specific T-cell responses are related to viral clearance, impaired responses can lead to chronic infection, turning HCV variability into a major obstacle for vaccine development. In a recent work, Fytili et al. (2008) studied the cross reactive potential of HCV specific CD8+ T-cells and observed a large variation in immunogenicity among 28 naturally occurring NS3(1073) variants. In this work, we intend to evaluate this immunogenic variation at molecular level, through bioinformatics approaches. The D1-EM-D2 strategy was used to build in silico MHC:peptide complexes (pMHC) of these HCV-derived peptides in the context of HLA-A*02:01 allele. The TCR-interacting surface of these complexes were evaluated using the GRASP2 program. Structural analysis indicated a sharing of topological and electrostatic features among complexes that induced strong response in vitro. Besides, complexes that induced low response presented an important positively charged spot in the center of TCR-interacting area. This spot was seen even in complexes with conservative amino acid changes and is consistent with the impairment of recognition by wild-type-specific T-cells, observed in vitro. Furthermore, the most remarkable difference in electrostatic potential was seen precisely in the only complex unable to induce in vitro stimulation. All these observations were confirmed by Principal Component Analysis (PCA) and this approach was also applied to a set of 45 non-related immunogenic viral epitopes, indicating possible new targets for cross-reactivity studies. Our results suggest structural in silico analysis of pMHC complexes as a reliable tool for vaccine development, affording to predict the impact of viral escape mutations and selection of epitopes with potential to induce cross-reactive immune responses.

Asunto(s)

Antígenos HLA-A/genética , Antígenos HLA-A/inmunología , Hepacivirus/genética , Hepacivirus/inmunología , Proteínas no Estructurales Virales/inmunología , Alelos , Secuencia de Aminoácidos , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Biología Computacional , Simulación por Computador , Hepatitis C/genética , Hepatitis C/inmunología , Humanos , Inmunidad Celular , Modelos Moleculares , Análisis de Componente Principal , Receptores de Antígenos de Linfocitos T/inmunología , Electricidad Estática , Proteínas no Estructurales Virales/química , Proteínas no Estructurales Virales/genética

RESUMEN

Este trabalho teve como objetivo verificar a ocorrência de Listeria monocytogenes em salames artesanais, elaborados por pequenas indústrias do setor de carnes do sul do Brasil. Das cinqüenta amostras de salames de diferentes fabricantes avaliadas, em 8% foi verificada a presença de L. monocytogenes. Os valores de pH dessas amostras variaram entre 4,35 e 5,99 e a atividade de água de 0,89 a 0,91. A implantação das Boas Práticas de Fabricação neste setor é fundamental para obter alimentos prontos para o consumo e seguros à saúde do consumidor.(AU)

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The objective of this work was to verify the occurrence of Listeria monocytogenes in artisanal sausages,elaborated in small scale meat industries from south of Brazil. Fifty samples of sausages from differentmanufacturers were evaluated, and the presence of L. monocytogenes was verified in 8% of the samples. Thevalues of pH of these samples varied between 4,35 and 5,99 and the water activity ranged from 0,89 to 0,91. The implantation of Good Manufacture Practices in this sector is basic to obtain foods ready to eat and safe to the consumer's health. (AU)

Asunto(s)

Productos de la Carne/microbiología , Microbiología de Alimentos , Contaminación de Alimentos , Muestras de Alimentos , Listeria monocytogenes/aislamiento & purificación , Listeria monocytogenes/patogenicidad , Brasil

RESUMEN

Este trabalho teve como objetivo verificar a ocorrência de Listeria monocytogenes em salames artesanais, elaborados por pequenas indústrias do setor de carnes do sul do Brasil. Das cinqüenta amostras de salames de diferentes fabricantes avaliadas, em 8% foi verificada a presença de L. monocytogenes. Os valores de pH dessas amostras variaram entre 4,35 e 5,99 e a atividade de água de 0,89 a 0,91. A implantação das Boas Práticas de Fabricação neste setor é fundamental para obter alimentos prontos para o consumo e seguros à saúde do consumidor.

Asunto(s)

Contaminación de Alimentos , Microbiología de Alimentos , Muestras de Alimentos , Productos de la Carne/microbiología , Brasil , Listeria monocytogenes/aislamiento & purificación , Listeria monocytogenes/patogenicidad

RESUMEN

Biological processes for wastewater treatment generally produce biomass or active sludge without reuse. In this context, incorporation of organic matter and nutrients from agro industrial effluents into cell mass for single-cell protein allowed application of sustainable process. Cyanobacteria could be used due to its versatile metabolism. So, the aim of this paper was evaluate the growth of cyanobacteria Aphanothece microscopica Nägeli growth on heterotrophic medium with glucose, lactose and sucrose. Growth curves indicated that cultivation of cyanobacterial on the dark depend the type of carbon source and there are different mechanisms for glucose, fructose and sucrose consumption. Results suggest a useful application of cyanobacteria on organic matter removal from wastewater.

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Os processos biológicos de tratamento de águas residuárias produzem grandes quantidades de biomassa geralmente sem utilização posterior. Neste contexto, a incorporação de matéria orgânica e nutrientes de efluentes agroindustriais em células microbianas visando a produção de proteínas unicelulares corresponderia a um processo sustentável. Nesse sentido, as cianobactérias poderiam ser aplicadas devido ao seu metabolismo versátil. Sendo assim, o trabalho teve como objetivo avaliar o cultivo heterotrófico da cianobactéria Aphanothece microscopica Nägeli em meios contendo glicose, lactose e sacarose. As curvas de crescimento indicaram que o cultivo heterotrófico depende do tipo de fonte de carbono, sugerindo diferentes mecanismos de incorporação e consumo da glicose, lactose sacarose. Os resultados indicam uma possível aplicação desta cianobactéria na remoção destas moléculas orgânicas em águas residuárias.

Asunto(s)

Cianobacterias , Procesos Heterotróficos , Purificación del Agua

RESUMEN

The immune system is engaged in a constant antigenic surveillance through the Major Histocompatibility Complex (MHC) class I antigen presentation pathway. This is an efficient mechanism for detection of intracellular infections, especially viral ones. In this work we describe conformational patterns shared by epitopes presented by a given MHC allele and use these features to develop a docking approach that simulates the peptide loading into the MHC cleft. Our strategy, to construct in silico MHC:peptide complexes, was successfully tested by reproducing four different crystal structures of MHC-I molecules available at the Protein Data Bank (PDB). An in silico study of cross-reactivity potential was also performed between the wild-type complex HLA-A2-NS31073 and nine MHC:peptide complexes presenting alanine exchange peptides. This indicates that structural similarities among the complexes can give us important clues about cross reactivity. The approach used in this work allows the selection of epitopes with potential to induce cross-reactive immune responses, providing useful tools for studies in autoimmunity and to the development of more comprehensive vaccines.

Asunto(s)

Presentación de Antígeno , Reacciones Cruzadas/inmunología , Epítopos/química , Antígenos de Histocompatibilidad Clase I/química , Complejo Mayor de Histocompatibilidad , Alelos , Biología Computacional/métodos , Simulación por Computador , Bases de Datos de Proteínas , Antígenos de Histocompatibilidad Clase I/metabolismo , Humanos , Conformación Molecular , Unión Proteica

RESUMEN

This study evaluated the biodegradation of catechol by a yeast strain of Candida parapsilopsis in standard medium in Erlenmeyer flasks. Results shown that the highest concentration of catechol caused the longer lag period, demonstrating that acclimatized cultures could completely degrade an initial catechol concentration of 910 mg/L within 48 h. Haldane's model validated the experimental data adequately for growth kinetics over the studied catechol concentration ranges of 36 to 910 mg/L. The constants obtained for this model were µmax = 0.246 h-1, Ks = 16.95 mg/L and Ki = 604.85 mg/L.

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Neste trabalho foi estudada a biodegradação de catecol em frascos de Erlenmeyers em água residuária sintética pela levedura Candida parapsilopsis. As respostas dos ensaios cinéticos mostraram que altas concentrações de catecol ocasionaram uma fase lag longa para a levedura. Portanto, a aclimatização da cultura de levedura empregada para biodegradação de catecol é de fundamental importância, sendo possível reduzir toda a concentração inicial de catecol da água residuária sintética de 910 mg/L em 48 horas. Os dados experimentais da cinética de biodegradação do catecol foram ajustados pelo modelo de Haldane adequadamente, sobre a faixa de concentração de catecol investigada de 36 a 910 mg/L. Os parâmetros cinéticos obtidos do modelo de Haldane foram: µmax = 0,246 h-1, Ks = 16,95 mg/L e Ki = 604,85 mg/L.