Higher Production Of Proinflammatory Cytokines Research Articles

Limonin ameliorates dextran sulfate sodium-induced chronic colitis in mice by inhibiting PERK-ATF4-CHOP pathway of ER stress and NF-κB signaling

Inflammatory bowel disease (IBD) is a chronic gastrointestinal inflammation regulated by intricate mechanisms. Limonin, a natural tetracyclic triterpenoid compound, possesses multiple bioactivities including anti-inflammation, anti-cancer and so on. However, the therapeutic potential and the underlying mechanism of limonin on IBD remain unclear. Here, we probe into the effect of limonin on chronic colitis induced by dextran sulfate sodium (DSS) and illustrated the potential mechanisms. We found that limonin relieved the risk and severity of DSS-induced chronic colitis in mice through various aspects including increasing body weight and colon length, decreasing the mortality rate, inhibiting MPO activity and improving colon pathology. Limonin also decreased the production of proinflammatory cytokines TNF-α, IL-1β, IL-6 and the expression of inflammatory proteins COX-2, iNOS in colon tissues from DSS-induced colitis mice. Moreover, limonin attenuated DSS-induced chronic colitis by inhibiting PERK-ATF4-CHOP pathway of endoplasmic reticulum (ER) stress and NF-κB signaling. In vitro, limonin not only decreased LPS-induced higher production of pro-inflammatory cytokines and inflammatory proteins mentioned above by inhibiting NF-κB signaling in macrophage cells RAW264.7, but also suppressed PERK-ATF4-CHOP pathway of ER stress. In summary, our study demonstrated that limonin mitigated DSS-induced chronic colitis via inhibiting PERK-ATF4-CHOP pathway of ER stress and NF-κB signaling. All of this study provides the possibility for limonin as an effective drug for chronic colitis of IBD in the future.

DIABETES MELLITUS, OBESIDADE E MAUS HÁBITOS DE VIDA: SUA RELAÇÃO COM A COVID-19

No ano de 2019, foi descoberto o vírus que viria a se chamar de SARS-CoV-2 e que provocou uma crise sanitária global. A pandemia de 2020, causada pela doença nomeada de Coronavírus disease 2019 ou COVID-19 alastrou-se rapidamente ao redor do mundo, impactando a população global e, especialmente, alguns grupos de risco, como pacientes com doenças cardiovasculares, obesos e diabéticos, por exemplo. Este estudo possui o objetivo de analisar e relatar as relações existentes entre a COVID-19 e os pacientes que possuem distúrbios metabólicos como a diabetes mellitus ou sofrem de obesidade, e ainda expor como hábitos de vida podem interferir na infecção do SARS-CoV-2. Trata-se de um levantamento bibliográfico sobre os principais consensos e estudos sobre a COVID-19. Sabe-se que o vírus inicia o processo de infecção no trato respiratório superior para dar início ao ciclo de replicação viral. A diabetes mellitus é considerada um fator de risco de alta gravidade e mortalidade, uma vez que a hiperglicemia altera a resposta imunológica do indivíduo, resultando em maiores riscos de infecções. Igualmente, pessoas obesas possuem um risco significativo devido à alta produção de citocinas pró-inflamatórias e alterações funcionais das células T que fazem ter uma resposta imunológica debilitada para conter a infecção. É essencial investir em medidas de prevenção de doenças, como a prática de atividades físicas e mudanças para bons hábitos alimentares com objetivo de melhorar a qualidade de vida e ter uma resposta imune eficaz especialmente no contexto pandêmico.

Mechanism of stress heart damage in the animals of different sensitivity to hypoxia

Hypoxia and resistance to it is the main in today data sitiuayion in the world.The aim of the study was to determine level of myocardium damage and corresponding of it with cytokines level in high- and low-resistance to hypoxic hypoxia rats (HR, LR) in different models of immobilization stress.Material and methods of investigation. The experiments were performed on 144 outbred HR and LR aged 5.5-6 months, dividing into 3 groups – control and 2 with different model of immobilizing stress. It was determined morphological changes of myocardium and concentration of interleukins 1beta, 2, 4, 6, 10 (IL-1beta, IL-2, IL-4, IL-6, IL-10), tumor necrosis factor alpha (TNF-alpha) in the blood serum.Results. The morphological investigation showed the bigger damage effect in LR animal, than in HR rats. Damage effects were higher in males, than in females. The stress repeating every 72 hours (stress 2) had higher damage in myocardium, than stress repeating every 24 hours (stress 1). We corresponds the results with cytokines level. High congenital resistance to hypoxia is associated with an increased content of anti-inflammatory cytokines (in males IL-4, in females IL-10). Intact males, compared with females, have a higher production of pro-inflammatory cytokines (IL-6, TNF-alpha).At stress1 inHR males, compared with HR females, were lower contents of IL-10, higher level of IL-1beta, IL-4, IL-6, TNF-alpha. At stress1 inLR males, compared with LR females, were lower contends of IL-10, IL-2. The morphological changes were more in males, that’s why higher level of anti-inflammatory cytokines IL-10 really has protection in HR and LR females’ animals. At stress2 inHR male, compared with HR female, were lower contents of IL-1beta and IL-10, higher IL-2 and IL-6. At stress2 inLR males, compared with LR females, were higher level of IL-10, IL-6, lower contents of IL-2, IL-4. The morphological changes were more in males, that’s why pro-inflammatory cytokines IL-6 really take place in damage of myocardium in males with different resistance animals.Conclusion. High congenital resistance to hypoxia is associated with smaller destroying of the myocardium layer in immobilizing stress, which correspond with increased content of anti-inflammatory cytokines IL-4 and, IL-10. Higher damage of myocardium in immobilizing stress correspond with higher production of pro-inflammatory cytokines (IL-6, TNF-alpha).

Chronic Tobacco Exposure by Smoking Develops Insulin Resistance.

The present review critically discusses the high occurrence rate, insulin resistance and type-2 diabetes in tobacco exposed individuals. Tobacco extracts and smoke contain a large number of toxic materials and a significant number of those are metabolic disintegrators. Glucose and lipid homeostasis is severely impaired by this compound. Tobacco exposure contributes to adverse effects by impairing the physical, biochemical and molecular mechanisms in the tissues. The immunological components are damaged by tobacco with high production of proinflammatory cytokines (IL-6, TNF-∞) and augmentation of inflammatory responses. These events result in damages to cytoskeletal structures of different tissues. Degradation of matrix structure (by activation of different types of MMPs) results in the permanent damages to the tissues and their metabolic functions. Cellular antioxidant defense system mostly cannot or hardly nullify CS-induced ROS production that activates polymorphonuclear neutrophils (PMNs), which are a major source of cytokines and chemokines (TNFα, IL6, IL8, INFγ). Additive effects of these immediately promote the low energy-metabolism as well as inflammation. Oxidative stress, mitochondrial dysfunction, and inflammation contribute to the direct nicotine toxicity via nAChRs in diabetes. The investigator identified that skeletal muscle insulin-resistance occurs in smokers due to phosphorylation of insulin receptor substrate1 (IRS1) at Ser-636 position. Tobacco exposure initiates free radical related immunological impairment, DNA damage, and inflammation. So, the present analysis is of importance to figure out the mechanistic layout of tobacco-induced tissue damage and its possible therapeutic interventions.

Obesity and Higher Risk for Severe Complications of Covid-19: What to do when the two pandemics meet.

The coronavirus disease 2019 (COVID-19) pandemic has spread around the globe, infecting more than ten million individuals, with more than 500,000 dead; about one half of the infected people have recovered. Despite this fact, a subgroup of individuals affected by COVID-19 is at greater risk of developing worse outcomes and experience a high rate of mortality. Data on the association between obesity and COVID-19 are growing; the available studies, have reported a high prevalence of overweight and obesity in patients experiencing a severe COVID-19 course, with serious complications requiring hospitalization and admission to intensive care units. This paper attempts to highlight potential mechanisms behind the greater vulnerability to COVID-19 of individuals with obesity. The presence of uncontrolled chronic obesity-related comorbidities, particularly pulmonary diseases, can present a primary fertile soil for respiratory tract infection. Combined with immune system impairments, such as alteration in the T-cell proliferation and macrophage differentiation, and the high pro-inflammatory cytokine production by the adipose organ, this may worsen the general condition toward a systemic diffusion of infection. Prevention remains the first line of intervention in these patients that can be achieved by adhering to social distancing and adopting hygiene precautions, combined with a healthy lifestyle. Patients with obesity require preferential access dedicated to primary care services to ensure they are regularly taking their medications for the treatment of any concurrent chronic diseases. Finally, their physicians must promptly manage any medical signs or symptoms in the case of suspected severe acute respiratory syndrome coronavirus-2 (SARS-CoV2) infection to prevent the risk of severe outcomes.

Mycobacterium lepromatosis genome exhibits unusually high CpG dinucleotide content and selection is key force in shaping codon usage

Mycobacterium lepromatosis was identified as a causative agent for leprosy in the year 2008 in the United States and later more cases were identified in Canada, Singapore, Brazil, and Myanmar. It is known to cause diffuse lepromatosis leprosy among humans. Since it is invasive, the mortality rates are higher in comparison to the M. leprae. At genomic level, there exists 90.9% similarity between M. lepromatosis and M. leprae. Codon usage analysis based on analyses of 228 coding sequences (CDSs) of M. lepromatosis, revealed that the genome is GC rich. Among the total 16 dinucleotides, CpG dinucleotide possesses the highest dinucleotide frequency in M. lepromatosis, that is strikingly an unobvious observation since higher CpG is associated with higher proinflammatory cytokine production and NF-κB activation that eventually leads to high pathogenicity. To evade immune response, CpG content is generally less in pathogens. The unusually high CpG content can be explained by the fact that the nucleotide composition of M. lepromatosis is CG rich. Various forces interplay to shape codon usage pattern of any organism including selection; mutation, nucleotide composition as well as GC biased gene conversion. To understand the interplay between various forces; neutrality, parity, Nc-GC3 (Effective number of codons-GC content at 3rd position of the codon), aromaticity (AROMO) and the general average hydropathicity score (GRAVY) analyses have been carried out. The analyses revealed that selection force is the major contributory force. Along with the selection; mutation, nucleotide composition as well as GC biased gene conversion also play role in shaping codon usage bias in M. lepromatosis. This is the first report on the codon usage in M. lepromatosis.

Changes in the content of interleukins in the serum of male and female rates with different resistance to acute hypoxic hypoxia under immobilization stress

Attention of scientists of the world are study the features of increased resistance to hypoxia and stress, different development inflammation in them.The aim of the study was to trace the features of interleukins level in high- and low-resistance to hypoxic hypoxia rats (HR, LR) in different models of immobilization stress.Material and methods of investigation. The experiments were performed on 144 outbred HR and LR aged 5.5-6 months, dividing into 3 groups – control and 2 with different model of immobilizing stress. Determine concentration of interleukins 1beta, 2, 4, 6, 10 (IL-1beta, IL-2, IL-4, IL-6, IL-10), tumor necrosis factor alpha (TNF-alpha) in the blood serum.Results. High congenital resistance to hypoxia is associated with an increased content of anti-inflammatory cytokines (in males - IL-4 on 24.73% (p<0.001), in females - IL-10 on 37.36%; p<0.001). Intact males, compared with females, have a higher production of pro-inflammatory cytokines (IL-6, TNF-alpha). Intact males, compared with females, are dominated by the content of pro-inflammatory cytokines in the blood. In HR males, compared with females, a higher content of IL-6 (49.93%; p<0.01), TNF-alpha (10.85%; p<0.01), IL-4 (33.92%; p<0.001), lower - IL-1beta (37.91%; p<0.02) and IL-10 (93.06%; p<0.001). In LR males, compared with females, higher blood levels of IL-1beta (42.98%; p<0.001), IL-6 (54.32%; p<0.001), TNF-alpha (16,40%; p<0,002), the highest level of pro-inflammatory cytokines under stress with a frequency of every 24 hours. In HR males under these conditions, the level of IL-1beta increases (in 2.35 times; p<0.001), TNF-alpha (in 8.20 times; p<0.001). In exposed to stress every 72 hours in HR males, the level of TNF-alpha increases in 2.88 times (p<0.002), and in LR males in 5.18 times (p<0.001). At the same time, in HR females its level increases in 3.25 times (p<0.01), in LR females - in 5.73 times (p<0.01).Conclusion. The experiments revealed the changes of pro- and anti-inflammatory interleukins, which depends of sex, resistance to hypoxy and kinds of stress.

NK cell receptor NKG2D enforces proinflammatory features and pathogenicity of Th1 and Th17 cells.

NKG2D is a danger sensor expressed on different subsets of innate and adaptive lymphocytes. Despite its established role as a potent activator of the immune system, NKG2D-driven regulation of CD4+ T helper (Th) cell-mediated immunity remains unclear. In this study, we demonstrate that NKG2D modulates Th1 and proinflammatory T-bet+ Th17 cell effector functions in vitro and in vivo. In particular, NKG2D promotes higher production of proinflammatory cytokines by Th1 and T-bet+ Th17 cells and reinforces their transcription of type 1 signature genes, including Tbx21. Conditional deletion of NKG2D in T cells impairs the ability of antigen-specific CD4+ T cells to promote inflammation in vivo during antigen-induced arthritis and experimental autoimmune encephalomyelitis, indicating that NKG2D is an important target for the amelioration of Th1- and Th17-mediated chronic inflammatory diseases.

TLR2 and Dectin-1 Signaling in Mouse Hematopoietic Stem and Progenitor Cells Impacts the Ability of the Antigen Presenting Cells They Produce to Activate CD4 T Cells

Microbial recognition by pattern recognition receptors (PRRs) expressed on hematopoietic stem and progenitor cells (HSPCs) not only activates myelopoiesis but also programs the function of the monocytes and macrophages they produce. For instance, changes in HSPC programming modify the ability of macrophages derived from them to produce inflammatory cytokines. While HSPCs exposed to a TLR2 agonist give rise to tolerized macrophages (lower proinflammatory cytokine production), HSPCs treated with Dectin-1 ligands produce trained macrophages (higher proinflammatory cytokine production). However, nothing is known about the impact of HSPC exposure to microbes on the function of antigen presenting cells (APCs). In this study we evaluated whether treatment of murine bone marrow HSPCs with a TLR2 or Dectin-1 ligand impacts the antigen presenting capacity of APCs derived from them in vitro. Following activation with microbial ligands or Candida albicans yeasts, APCs derived from TLR2/Dectin-1-programed HSPCs exhibit altered expression of MHCII (signal 1), co-stimulatory molecules (CD40, CD80 and CD86; signal 2) and cytokines (TNF-α, IL-6, IL-12 p40 and IL-2; signal 3). Moreover, APCs derived from TLR2/Dectin-1-programed HSPCs prime enhanced Th1 and Th17 responses, which are important for antifungal defense, in CD4 T cell cocultures. Overall, these results demonstrate for the first time that microbial detection by bone marrow HSPCs can modulate the adaptive immune response by inducing the production of APCs with an altered phenotype.

Induction of systemic immunity through nasal-associated lymphoid tissue (NALT) of mice intranasally immunized with Brucella abortus malate dehydrogenase-loaded chitosan nanoparticles.

Infection with Brucella abortus causes contagious zoonosis, brucellosis, and leads to abortion in animals and chronic illness in humans. Chitosan nanoparticles (CNs), biocompatible and nontoxic polymers, acts as a mucosal adjuvant. In our previous study, B. abortus malate dehydrogenase (Mdh) was loaded in CNs, and it induced high production of pro-inflammatory cytokines in THP-1 cells and systemic IgA in BALB/C mice. In this study, the time-series gene expression analysis of nasal-associated lymphoid tissue (NALT) was performed to identify the mechanism by which Mdh affect the target site of nasal immunization. We showed that intranasal immunization of CNs-Mdh reduced cell viability of epithelial cells and muscle cells at first 1 h, then induced cellular movement of immune cells such as granulocytes, neutrophils and lymphocytes at 6h, and activated IL-6 signaling pathway at 12h within NALT. These activation of immune cells also promoted signaling pathway for high-mobility group box 1 protein (HMGB1), followed by the maturation of DCs required for mucosal immunity. The CNs also triggered the response to other organism and inflammatory response, showing it is immune-enhancing adjuvant. The ELISA showed that significant production of specific IgA was detected in the fecal excretions and genital secretions from the CNs-Mdh-immunized group after 2 weeks-post immunization. Collectively, these results suggest that B. abortus Mdh-loaded CNs triggers activation of HMGB1, IL-6 and DCs maturation signaling within NALT and induce production of systemic IgG and IgA.

Induction of Th2 response through TLR2-mediated MyD88-dependent pathway in human microfold cells stimulated with chitosan nanoparticles loaded with Brucella abortus Mdh

Drug delivery by the nasal or oral route is considered the preferred route of administration because it can induce systemic mucosal immunity. However, few studies have examined the immunogenicity and transport of antigen at the level of the microfold (M) cell, the epithelial cell that specializes in antigen sampling at mucosal surfaces. In our previous study, Brucella abortus malate dehydrogenase (Mdh) was loaded in chitosan nanoparticles (CNs), and it induced high production of proinflammatory cytokines in THP-1 cells and systemic IgA in BALB/C mice. In the present study, an in vitro M cell model was used in which Caco-2 cells and Raji B cells were co-cultured to investigate the impact of the uptake and immunogenicity of B. abortus Mdh on nanoparticle transport in human M cells. Our results showed that loaded CNs induced enhanced transport of Mdh in the M cell model. ELISAs showed significantly higher production of IL-1β and IL-6 in the CN-Mdh stimulation group than that seen in the Mdh stimulation group. The observed increase of gene expression of TLR2, MyD88, TRAF6, IRF4 and CD14 implied that MyD88-dependent TLR2 signaling was activated by stimulation with CNs-Mdh. These results suggest that Mdh and CNs may function synergistically to enhance Th2-related responses triggered by the MyD88-dependent TLR2 signaling pathway and could induce an inflammatory response in M cells as an M cell-targeted delivery system. This study will contribute to the development of not only effective antigens for intracellular bacteria, including B. abortus, but also vaccine delivery systems that target M cells.

Depressive-like phenotype evoked by lifelong nutritional omega-3 deficiency in female rats: Crosstalk among kynurenine, Toll-like receptors and amyloid beta oligomers

Depression is one of the most common psychiatric diseases and the prevalence of depressive symptoms in women is almost twice compared to men, although the reasons of this gender difference are not fully understood yet. Recently, soluble amyloid beta (Aβ)1-42 peptide has been receiving great importance in the development of depression, also considering that depression is highly comorbid with Alzheimer’s disease and other neurodegenerative illnesses. The central role played by Aβ in the development of depressive-like symptoms in rodents has been evidenced in environmental rodent model of depression. Indeed, we have previously found that lifelong exposure to n-3 polyunsaturated fatty acids (PUFA) deficient diet in female rats at 8 weeks of life leads to depressive like- symptoms and higher susceptibility to stress associated with increased Aβ levels. In order to understand if such effects were maintained over time, rats were exposed to the same diet regimen until 6 or 21 weeks of life. We found that both timepoints of exposure to n-3 PUFA deficient diet lead to depressive-like phenotype. Furthermore, a significant alteration in brain neurochemistry was retrieved. In particular, in hippocampal area a significant reduction in serotonin (5-HT) and noradrenaline (NA) content was evidenced. Considering the prominent role of NA in counterbalancing neuroinflammatory state, we quantified in the same brain area kynurenine levels, a metabolite of tryptophan implicated in inflammatory state and brought to the fore for its implication in depression. Interestingly, kynurenine levels were significantly increased in hippocampus (HIPP) of female rats exposed to such diet. In addition, lifelong deficiency in n-3 PUFA dietary intake led to systemic increase of corticosterone, hence hypothalamic pituitary adrenal (HPA) axis hyperactivation, and higher proinflammatory cytokine production. Increased production of kynurenine, along with HPA axis hyperactivation, have been associated with immune system modulation, particularly through Toll-like receptor type 2 (TLR2) and Toll-like receptor type 4 (TLR4) involvement. In addition, it has been shown that soluble forms of Aβ1-42 can induced depressive like-phenotype in consequence to a crosstalk between TLR4 and 5-HTergic system. Thus, considering that in this model we have previously reported increased plasma Aβ1-42 level, we quantified TRL2 and 4 expression in HIPP of treated rats. We found that chronic exposure to a diet characterized by very low n-3 PUFA content led to higher expression of TLR2 and TLR4 in HIPP of female treated rats, indicating an activation of the immune system and was accompanied by increased expression of oligomeric Aβ.Taken together, our data indicate that the pro-depressive effects induced by a diet poor in n-3 PUFA can be attributable to a shift of hippocampal tryptophan metabolism toward inflammatory metabolite ultimately corresponding to altered immune response and increased Aβ oligomerization.

LL-37 alone and in combination with IL17A enhances proinflammatory cytokine expression in parallel with hyaluronan metabolism in human synovial sarcoma cell line SW982-A step toward understanding the development of inflammatory arthritis.

LL-37 is the only human cathelicidin-family host defense peptide and has been reported to interact with invading pathogens causing inflammation at various body sites. Recent studies showed high levels of LL-37 in the synovial-lining membrane of patients with rheumatoid arthritis, a common type of inflammatory arthritis. The present study aims to investigate the role of LL-37 on mechanisms associated with pathogenesis of inflammatory arthritis. The effects of LL-37 on the expression of proinflammatory cytokines, hyaluronan (HA) metabolism-related genes, cell death-related pathways, and cell invasion were investigated in SW982, a human synovial sarcoma cell line. Time-course measurements of proinflammatory cytokines and mediators showed that LL-37 significantly induced IL6 and IL17A mRNA levels at early time points (3–6 hr). HA-metabolism-related genes (i.e., HA synthase 2 (HAS2), HAS3, hyaluronidase 1 (HYAL1), HYAL2, and CD44) were co-expressed in parallel. In combination, LL-37 and IL17A significantly enhanced PTGS2, TNF, and HAS3 gene expression concomitantly with the elevation of their respective products, PGE2, TNF, and HA. Cell invasion rates and FN1 gene expression were also significantly enhanced. However, LL-37 alone or combined with IL17A did not affect cell mortality or cell cycle. Treatment of SW982 cells with both LL-37 and IL17A significantly enhanced IKK and p65 phosphorylation. These findings suggest that the chronic production of a high level of LL-37 may synchronize with its downstream proinflammatory cytokines, especially IL17A, contributing to the co-operative enhancement of pathogenesis mechanisms of inflammatory arthritis, such as high production of proinflammatory cytokines and mediators together with the activation of HA-metabolism-associated genes and cell invasion.

A T164S mutation in the dengue virus NS1 protein is associated with greater disease severity in mice.

Dengue viruses cause severe and sudden human epidemics worldwide. The secreted form of the nonstructural protein 1 (sNS1) of dengue virus causes vascular leakage, a hallmark of severe dengue disease. Here, we reverse engineered the T164S mutation of NS1, associated with the severity of dengue epidemics in the Americas, into a dengue virus serotype 2 mildly infectious strain. The T164S mutant virus decreased infectious virus production and increased sNS1 production in mammalian cell lines and human peripheral blood mononuclear cells (PBMCs) without affecting viral RNA replication. Gene expression profiling of 268 inflammation-associated human genes revealed up-regulation of genes induced in response to vascular leakage. Infection of the mosquito vector Aedes aegypti with the T164S mutant virus resulted in increased viral load in the mosquito midgut and higher sNS1 production compared to wild-type virus infection. Infection of type 1 and 2 interferon receptor-deficient AG129 mice with the T164S mutant virus resulted in severe disease coupled with increased complement activation, tissue inflammation, and more rapid mortality compared to AG129 mice infected with wild-type virus. Molecular dynamics simulations predicted that mutant sNS1 formed stable dimers similar to the wild-type protein, whereas the hexameric mutant sNS1 was predicted to be unstable. Immunoaffinity-purified sNS1 from T164S mutant virus-infected mammalian cells was associated with different lipid classes compared to wild-type sNS1. Treatment of human PBMCs with sNS1 purified from T164S mutant virus resulted in a twofold higher production of proinflammatory cytokines, suggesting a mechanism for how mutant sNS1 may cause more severe dengue disease.

Influence of Preparation and Exposure Periods of Eluates from Ocular Prosthesis Acrylic Resin in Human Conjunctival Cell Line

This study was undertaken to analyze if different preparation and exposure periods of eluates from ocular prosthesis acrylic resin influence the cytotoxicity for conjunctival cells. Twenty-four acrylic resin specimens were divided, according to the period of eluate exposure to Chang conjunctival cells (24 and 72 hours). Eluates were prepared in four different ways: 24, 48, and 72 hours of resin specimen immersion in medium and 24 hours of immersion in water, followed by 24 hours of immersion in medium. MTT assay was used to evaluate the cytotoxic effect. The production of IL-1β, IL-6, TNF-α, and chemokine macrophage inflammatory protein 1α was evaluated by ELISA, while the mRNA expression of type IV collagen (COL IV), transforming growth factor β (TGF-β), and matrix metalloproteinase 9 (MMP9) were evaluated by real-time RT-PCR technique. The statistical analysis was carried out using ANOVA with Bonferroni post-hoc test and the student’s t-test (p < 0.05). Significant quantities of IL-6 (4.594 pg/mL) and mRNA expression of COL IV (1.58) were verified at 72 hours of eluate exposure to cells, as compared to 24 hours. After the 72-hour exposure of eluates to cells, lower cell proliferation (88.4%) and higher IL-6 quantities (12.374 pg/mL), as well as mRNA expression of COL IV (2.21), TGF-β (2.02), and MMP9 (5.75) were observed, which corresponded to 72 hours of a specimen immersed in medium. Longer periods of eluate preparation and exposure from the acrylic resin to cells are related to higher production of proinflammatory cytokines and extracellular matrix proteins.

Influence of Preparation and Exposure Periods of Eluates from Ocular Prosthesis Acrylic Resin in Human Conjunctival Cell Line

Background:This study was undertaken to analyze if different preparation and exposure periods of eluates from ocular prosthesis acrylic resin influence the cytotoxicity for conjunctival cells.Methods:Twenty-four acrylic resin specimens were divided, according to the period of eluate exposure to Chang conjunctival cells (24 and 72 hours). Eluates were prepared in four different ways: 24, 48, and 72 hours of resin specimen immersion in medium and 24 hours of immersion in water, followed by 24 hours of immersion in medium. MTT assay was used to evaluate the cytotoxic effect. The production of IL-1β, IL-6, TNF-α, and chemokine macrophage inflammatory protein 1α was evaluated by ELISA, while the mRNA expression of type IV collagen (COL IV), transforming growth factor β (TGF-β), and matrix metalloproteinase 9 (MMP9) were evaluated by real-time RT-PCR technique. The statistical analysis was carried out using ANOVA with Bonferroni post-hoc test and the student’s t-test (p < 0.05).Results:Significant quantities of IL-6 (4.594 pg/mL) and mRNA expression of COL IV (1.58) were verified at 72 hours of eluate exposure to cells, as compared to 24 hours. After the 72-hour exposure of eluates to cells, lower cell proliferation (88.4%) and higher IL-6 quantities (12.374 pg/mL), as well as mRNA expression of COL IV (2.21), TGF-β (2.02), and MMP9 (5.75) were observed, which corresponded to 72 hours of a specimen immersed in medium.Conclusion:Longer periods of eluate preparation and exposure from the acrylic resin to cells are related to higher production of proinflammatory cytokines and extracellular matrix proteins.

Peculiarities of phenotypes of patients with pyelo- and glomerulonephritis by HLA distribution analysis

Studies devoted to the role of human leucocyte antigens (HLA) in pathogenesis of chronic kidney disease (CKD) have demonstrated the associative links of the HLA antigens, which stipulate the relative and attributive risks of some autoimmune diseases, with immune disorder and a high production of pro-inflammatory cytokines.&#x0D; The aim of our study was to determine the peculiarities of phenotypes of CKD patients according to the distribution of HLA-A, B and DR antigens and to conduct their comparative analysis in patients with pyelonephritis (PN) and glomerulonephritis (GN).&#x0D; Methods: The distribution of HLA-A, B, DR antigens in 384 CKD patients (120 with PN and 264 with GN) was analyzed. HLA antigens were defined using a standard microlymphocytotoxic test on the Terasakiґs planchette with special panels of anti-HLA serums (20 antigens of locus A, 31 – B and 9 – DR). The control group consisted of 350 healthy donors.&#x0D; The HLA antigen frequencies in normal and diseased subjects were compared taking each antigen separately, using χ2 test. The etiologic fraction (attributive risk s &gt; 0,1) was counted using the formula: s = x - y/I- y, where x is frequency of antigen in patients and y is frequency in healthy. The s reading was considered reliable when it exceeded 0.1.&#x0D; Results. The causal role (σ &gt; 0,1) was determined for А10, А11; В14, В16 for PN; antigens-protectors - А2, В21, В35, В40.&#x0D; For CGN, NS the relative risk is high (RR &gt; 2) at the presence of HLA-A23, А24, А28; B8, В38, В41, В44; DR1, DR4, DRw52 in phenotype, the causal role in etiopathology (σ&gt;0.1) is indicated for A24,А28; B8; DR1, DR4, DRw52; the disease protectors are B12 and B16.&#x0D; Conclusion. Conclusion. The features of the HLA-phenotype of patients with pyelo- and glomerulonephritis were shown. It allowed to establish the interconnectedness of the antigens of the histocompatibility complex with the risk of kidney diseases developing, which could help to personificate of the treatment and predicte of the course of the disease.

Early Senescence and Leukocyte Telomere Shortening in SCHIZOPHRENIA: A Role for Cytomegalovirus Infection?

Schizophrenia is a severe, chronic mental disorder characterized by delusions and hallucinations. Several evidences support the link of schizophrenia with accelerated telomeres shortening and accelerated aging. Thus, schizophrenia patients show higher mortality compared to age-matched healthy donors. The etiology of schizophrenia is multifactorial, involving genetic and environmental factors. Telomere erosion has been shown to be accelerated by different factors including environmental factors such as cigarette smoking and chronic alcohol consumption or by psychosocial stress such as childhood maltreatment. In humans, telomere studies have mainly relied on measurements of leukocyte telomere length and it is generally accepted that individuals with short leukocyte telomere length are considered biologically older than those with longer ones. A dysregulation of both innate and adaptive immune systems has been described in schizophrenia patients and other mental diseases supporting the contribution of the immune system to disease symptoms. Thus, it has been suggested that abnormal immune activation with high pro-inflammatory cytokine production in response to still undefined environmental agents such as herpesviruses infections can be involved in the pathogenesis and pathophysiology of schizophrenia. It has been proposed that chronic inflammation and oxidative stress are involved in the course of schizophrenia illness, early onset of cardiovascular disease, accelerated aging, and premature mortality in schizophrenia. Prenatal or neonatal exposures to neurotropic pathogens such as Cytomegalovirus or Toxoplasma gondii have been proposed as environmental risk factors for schizophrenia in individuals with a risk genetic background. Thus, pro-inflammatory cytokines and microglia activation, together with genetic vulnerability, are considered etiological factors for schizophrenia, and support that inflammation status is involved in the course of illness in schizophrenia.

Interaction Between Macrophage Migration Inhibitory Factor and CD74 in Human Immunodeficiency Virus Type I Infected Primary Monocyte-Derived Macrophages Triggers the Production of Proinflammatory Mediators and Enhances Infection of Unactivated CD4+ T Cells

Understanding the mechanisms of human immunodeficiency virus type I (HIV-1) pathogenesis would facilitate the identification of new therapeutic targets to control the infection in face of current antiretroviral therapy limitations. CD74 membrane expression is upregulated in HIV-1-infected cells and the magnitude of its modulation correlates with immune hyperactivation in HIV-infected individuals. In addition, plasma level of the CD74 activating ligand macrophage migration inhibitory factor (MIF) is increased in infected subjects. However, the role played by MIF/CD74 interaction in HIV pathogenesis remains unexplored. Here, we studied the effect of MIF/CD74 interaction on primary HIV-infected monocyte-derived macrophages (MDMs) and its implications for HIV immunopathogenesis. Confocal immunofluorescence analysis of CD74 and CD44 (the MIF signal transduction co-receptor) expression indicated that both molecules colocalized at the plasma membrane specifically in wild-type HIV-infected MDMs. Treatment of infected MDMs with MIF resulted in an MIF-dependent increase in TLR4 expression. Similarly, there was a dose-dependent increase in the production of IL-6, IL-8, TNFα, IL-1β, and sICAM compared to the no-MIF condition, specifically from infected MDMs. Importantly, the effect observed on IL-6, IL-8, TNFα, and IL-1β was abrogated by impeding MIF interaction with CD74. Moreover, the use of a neutralizing αMIF antibody or an MIF antagonist reverted these effects, supporting the specificity of the results. Treatment of unactivated CD4+ T-cells with MIF-treated HIV-infected MDM-derived culture supernatants led to enhanced permissiveness to HIV-1 infection. This effect was lost when CD4+ T-cells were treated with supernatants derived from infected MDMs in which CD74/MIF interaction had been blocked. Moreover, the enhanced permissiveness of unactivated CD4+ T-cells was recapitulated by exogenous addition of IL-6, IL-8, IL-1β, and TNFα, or abrogated by neutralizing its biological activity using specific antibodies. Results obtained with BAL and NL4-3 HIV laboratory strains were reproduced using transmitted/founder primary isolates. This evidence indicated that MIF/CD74 interaction resulted in a higher production of proinflammatory cytokines from HIV-infected MDMs. This caused the generation of an inflammatory microenvironment which predisposed unactivated CD4+ T-cells to HIV-1 infection, which might contribute to viral spreading and reservoir seeding. Overall, these results support a novel role of the MIF/CD74 axis in HIV pathogenesis that deserves further investigation.

High production of pro-inflammatory cytokines by maternal blood mononuclear cells is associated with reduced maternal malaria but increased cord blood infection

BackgroundIncreased susceptibility to malaria during pregnancy is not completely understood. Cellular immune responses mediate both pathology and immunity but the effector responses involved in these processes have not been fully characterized. Maternal and fetal cytokine and chemokine responses to malaria at delivery, and their association with pregnancy and childhood outcomes, were investigated in 174 samples from a mother and child cohort from Mozambique. Peripheral and cord mononuclear cells were stimulated with Plasmodium falciparum lysate and secretion of IL-12p70, IFN-γ, IL-2, IL-10, IL-8, IL-6, IL-4, IL-5, IL-1β, TNF, TNF-β was quantified in culture supernatants by multiplex flow cytometry while cellular mRNA expression of IFN-γ, TNF, IL-2, IL-4, IL-6, IL-10 and IL-13 was measured by quantitative PCR.ResultsHigher concentrations of IL-6 and IL-1β were associated with a reduced risk of P. falciparum infection in pregnant women (p < 0.049). Pro-inflammatory cytokines IL-6, IL-1β and TNF strongly correlated among themselves (ρ > 0.5, p < 0.001). Higher production of IL-1β was significantly associated with congenital malaria (p < 0.046) and excessive TNF was associated with peripheral infection and placental lesions (p < 0.044).ConclusionsComplex network of immuno-pathological cytokine mechanisms in the placental and utero environments showed a potential trade-off between positive and negative effects on mother and newborn susceptibility to infection.