46 results on '"Bonato VLD"'
Search Results
2. Tissue distribution of a plasmid DNA encoding Hsp65 gene is dependent on the dose administered through intramuscular delivery
- Author
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Coelho-Castelo, AAM, primary, Trombone, AP, additional, Rosada, RS, additional, Santos, RR, additional, Bonato, VLD, additional, Sartori, A, additional, and Silva, CL, additional
- Published
- 2006
- Full Text
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3. Characterization of the memory/activated T cells that mediate the long-lived host response against tuberculosis after bacillus Calmette-Guerin or DNA vaccination
- Author
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Silva, Cl, Bonato, Vld, Lima, Vmf, Faccioli, Lh, and Sylvia Leao
4. Alpha 1-acid glycoprotein is upregulated in severe COVID-19 patients and decreases neutrophil NETs in SARS-CoV-2 infection.
- Author
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Mestriner F, Francisco DF, Campos LCB, Couto AES, Fraga-Silva TFC, Flora Dugaich V, D Avila-Mesquita C, Zukowski Kovacs H, Vasconcelos JL, Milani ER, Santos Guedes de Sá K, Martins R, Jordani MC, Corsi CAC, Barbosa JM, Vasconcelos T, Gonçalves Menegueti M, Neto J, da Costa RM, Evora PRB, Arruda E, Tostes R, Polonis K, Bonato VLD, Auxiliadora-Martins M, Ribeiro MS, and Becari C
- Subjects
- Humans, Neutrophils metabolism, Orosomucoid metabolism, Orosomucoid pharmacology, SARS-CoV-2, Interleukin-6 metabolism, Receptor Protein-Tyrosine Kinases metabolism, Immunoproteins metabolism, COVID-19 metabolism
- Abstract
Orosomucoid, or alpha-1 acid glycoprotein (AGP), is a major acute-phase protein expressed in response to systemic injury and inflammation. AGP has been described as an inhibitor of neutrophil migration on sepsis, particularly its immunomodulation effects. AGP's biological functions in coronavirus disease 2019 (COVID-19) are not understood. We sought to investigate the role of AGP in severe COVID-19 infection patients and neutrophils infected with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Epidemiological data, AGP levels, and other laboratory parameters were measured in blood samples from 56 subjects hospitalized in the ICU with SARS-CoV-2 infection. To evaluate the role of AGP in NETosis in neutrophils, blood samples from health patients were collected, and neutrophils were separated and infected with SARS-CoV-2. Those neutrophils were treated with AGP or vehicle, and NETosis was analyzed by flow cytometry. AGP was upregulated in severe COVID-19 patients (p<0.05). AGP level was positively correlated with IL-6 and C-reactive protein (respectively, p=0.005, p=0.002) and negatively correlated with lactate (p=0.004). AGP treatment downregulated early and late NETosis (respectively, 35.7% and 43.5%) in neutrophils infected with SARS-CoV-2 and up-regulated IL-6 supernatant culture expression (p<0.0001). Our data showed increased AGP in COVID-19 infection and contributed to NETosis regulation and increased IL-6 production, possibly related to the Cytokine storm in COVID-19., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)
- Published
- 2024
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5. Different bacterial cargo in apoptotic cells drive distinct macrophage phenotypes.
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Salina ACG, de Aquino Penteado L, Dejani NN, Silva-Pereira L, Raimundo BVB, Corrêa GF, Oliveira KC, Ramalho LNZ, Boko MMM, Bonato VLD, Henrique Serezani C, and Medeiros AI
- Subjects
- Macrophages metabolism, Phenotype, Anti-Inflammatory Agents, Apoptosis, Phagocytosis
- Abstract
The removal of dead cells (efferocytosis) contributes to the resolution of the infection and preservation of the tissue. Depending on the environment milieu, macrophages may show inflammatory (M1) or anti-inflammatory (M2) phenotypes. Inflammatory leukocytes are recruited during infection, followed by the accumulation of infected and non-infected apoptotic cells (AC). Efferocytosis of non-infected AC promotes TGF-β, IL-10, and PGE
2 production and the polarization of anti-inflammatory macrophages. These M2 macrophages acquire an efficient ability to remove apoptotic cells that are involved in tissue repair and resolution of inflammation. On the other hand, the impact of efferocytosis of infected apoptotic cells on macrophage activation profile remains unknown. Here, we are showing that the efferocytosis of gram-positive Streptococcus pneumoniae-AC (Sp-AC) or gram-negative Klebsiella pneumoniae-AC (Kp-AC) promotes distinct gene expression and cytokine signature in macrophages. Whereas the efferocytosis of Kp-AC triggered a predominant M1 phenotype in vitro and in vivo, the efferocytosis of Sp-AC promoted a mixed M1/M2 activation in vitro and in vivo in a model of allergic asthma. Together, these findings suggest that the nature of the pathogen and antigen load into AC may have different impacts on inducing macrophage polarization., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)- Published
- 2024
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6. Effect of Dietary Supplementation with Omega-3 Fatty Acid on the Generation of Regulatory T Lymphocytes and on Antioxidant Parameters and Markers of Oxidative Stress in the Liver Tissue of IL-10 Knockout Mice.
- Author
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Campanari DD, Cipriano UG, Fraga-Silva TFC, Ramalho LNZ, Ovidio PP, Jordão Júnior AA, Bonato VLD, and Ferriolli E
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- Female, Mice, Animals, Antioxidants pharmacology, T-Lymphocytes, Regulatory metabolism, Mice, Knockout, Interleukin-10 metabolism, Mice, Inbred C57BL, Fish Oils pharmacology, Oxidative Stress, Dietary Supplements, Liver metabolism, Inflammation metabolism, Fatty Acids, Omega-3 pharmacology
- Abstract
Introduction: chronic low-grade inflammation, or inflammaging, emerges as a crucial element in the aging process and is associated with cardiovascular and neurological diseases, sarcopenia, and malnutrition. Evidence suggests that omega-3 fatty acids present a potential therapeutic agent in the prevention and treatment of inflammatory diseases, mitigating oxidative stress, and improving muscle mass, attributes that are particularly relevant in the context of aging. The objective of the present study was to evaluate the effectiveness of supplementation with omega-3 fish oil in improving the immune response and oxidative stress in knockout mice for interleukin IL-10 (IL-10
-/- )., Material and Methods: female C57BL/6 wild-type (WT) and interleukin IL-10 knockout (IL-10-/- ) mice were fed during 90 days with a standard diet (control groups), or they were fed/supplemented with 10% of the omega-3 polyunsaturated fatty acid diet (omega-3 groups). Muscle, liver, intestinal, and mesenteric lymph node tissue were collected for analysis., Results: the IL-10-/- +O3 group showed greater weight gain compared to the WT+O3 ( p = 0.001) group. The IL-10-/- +O3 group exhibited a higher frequency of regulatory T cells than the IL-10-/- group ( p = 0.001). It was found that animals in the IL-10-/- +O3 group had lower levels of steatosis when compared to the IL-10-/- group ( p = 0.017). There was even greater vitamin E activity in the WT group compared to the IL-10-/- +O3 group ( p = 0.001) and WT+O3 compared to IL-10-/- +O3 ( p = 0.002), and when analyzing the marker of oxidative stress, MDA, an increase in lipid peroxidation was found in the IL-10-/- +O3 group when compared to the IL-10-/- group ( p = 0.03). Muscle tissue histology showed decreased muscle fibers in the IL-10-/- +O3, IL-10-/- , and WT+O3 groups., Conclusion: the findings show a decrease in inflammation, an increase in oxidative stress markers, and a decrease in antioxidant markers in the IL-10-/- +O3 group, suggesting that supplementation with omega-3 fish oil might be a potential intervention for inflammaging that characterizes the aging process and age-related diseases.- Published
- 2024
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7. Inulin prebiotic ameliorates type 1 diabetes dictating regulatory T cell homing via CCR4 to pancreatic islets and butyrogenic gut microbiota in murine model.
- Author
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Guimarães JB, Rodrigues VF, Pereira ÍS, Manso GMDC, Elias-Oliveira J, Leite JA, Waldetario MCGM, de Oliveira S, Gomes ABDSP, Faria AMC, Ramos SG, Bonato VLD, Silva JS, Vinolo MAR, Sampaio UM, Clerici MTPS, and Carlos D
- Subjects
- Mice, Animals, Inulin pharmacology, Prebiotics, Disease Models, Animal, T-Lymphocytes, Regulatory, Butyrates pharmacology, Forkhead Transcription Factors, Diabetes Mellitus, Type 1, Gastrointestinal Microbiome, Islets of Langerhans
- Abstract
Gut dysbiosis is linked to type 1 diabetes mellitus (T1D). Inulin (INU), a prebiotic, modulates the gut microbiota, promoting beneficial bacteria that produce essential short-chain fatty acids for immune regulation. However, how INU affects T1D remains uncertain. Using a streptozotocin-induced (STZ) mouse model, we studied INU's protective effects. Remarkably, STZ + INU mice resisted T1D, with none developing the disease. They had lower blood glucose, reduced pancreatic inflammation, and normalized serum insulin compared with STZ + SD mice. STZ + INU mice also had enhanced mucus production, abundant Bifidobacterium, Clostridium cluster IV, Akkermansia muciniphila, and increased fecal butyrate. In cecal lymph nodes, we observed fewer CD4+Foxp3+ regulatory T cells expressing CCR4 and more Foxp3+CCR4+ cells in pancreatic islets, with higher CCL17 expression. This phenotype was absent in CCR4-deficient mice on INU. INU supplementation effectively protects against experimental T1D by recruiting CCR4+ regulatory T cells via CCL17 into the pancreas and altering the butyrate-producing microbiota., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2023. Published by Oxford University Press on behalf of Society for Leukocyte Biology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2024
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8. Divergent androgenic modulation of SARS-CoV-2 infection cooperates with dysregulated immune response to dictate worse COVID-19 outcomes in men.
- Author
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Duarte-Silva M, Oliveira CNS, Fuzo C, Silva-Neto PV, Toro DM, Pimentel VE, Pérez MM, Fraga-Silva TFC, Carvalho JCS, Neto FMS, Júnior RBM, Arruda E, Vilar FC, Degiovani AM, Ostini FM, Feitosa MR, Parra RS, Gaspar GG, Rocha JJR, Feres O, Fernandes APM, Maruyama SR, Russo EMS, Bonato VLD, Santos IKFM, Sorgi CA, Dias-Baruffi M, Faccioli LH, and Cardoso CRB
- Abstract
Background: Sex-determined differences are rarely addressed in the management of diseases, despite well-known contrasting outcomes between female and male patients. In COVID-19 there is a remarkable disparity, with higher rates of mortality and more severe acute disease in men compared to women, who are mostly affected by long COVID-19. Furthermore, whether androgens play a protective or detrimental role in COVID-19 is still a matter of debate. Hence, the adequate management of the disease, especially regarding men presenting acute disease aggravation, still needs important data to elucidate the interplay between sex hormones and host immune responses that drive the worse evolution in male patients., Methods: A cohort of 92 controls and 198 non-severe and severe COVID-19 patients, from both sexes, was assessed for clinical outcomes, plasma steroids, gonadotropins, sex hormone binding globulin (SHBG) and immune mediators, before vaccination. These data were correlated with the global gene expression of blood leukocytes. The androgen receptor (AR) signaling pathway was investigated by transcriptomics and tracheal aspirate was obtained from severe patients for SARS-COV-2 quantification in the respiratory tract. The interplay among clinical, endocrine and immunological data deciphered the sex differences in COVID-19. Importantly, statistical analyses, using 95% confidence interval, considered confounding factors such as age and comorbidities, to definitely parse the role of androgens in the disease outcome., Results: There were notable contrasting levels of testosterone and dihydrotestosterone (DHT) throughout the disease course in male but not female patients. Inflammatory mediators presented significant negative correlations with testosterone, which was partially dependent on age and diabetes in men. Male subjects with severe COVID-19 had a significant up regulation of the AR signaling pathway, including modulation of TMPRSS2 and SRD5A1 genes, which are related to the viral infection and DHT production. Indeed, men had a higher viral load in the tracheal aspirate and levels of DHT were associated with increased relative risk of death. In contrast, the testosterone hormone, which was notably reduced in severe disease, was significantly related with susceptibility to COVID-19 worsening in male patients. Secondary hypogonadism was ruled out in the male severe COVID-19 subjects, as FSH, LH, and SHBG levels were not significantly altered. Instead, these subjects tended to have increased gonadotropin levels. Most interestingly, in this study we identified, for the first time, combined sets of clinical and immunoendocrine parameters that together predicted progression from non-severe to severe COVID-19 in men. One of the limitations of our study was the low or undetectable levels of DHT in many patients. Then, the evaluation of enzymes related to biosynthesis and signaling by androgens was mandatory and reiterated our findings., Conclusions: These original results unraveled the disease immunoendocrine regulation, despite vaccination or comorbidities and pointed to the fundamental divergent role of the androgens testosterone and DHT in the determination of COVID-19 outcomes in men. Therefore, sex-specific management of the dysregulated responses, treatments or public health measures should be considered for the control of COVID-19 pandemic., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)
- Published
- 2023
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9. Airway epithelial cells and macrophages trigger IL-6-CD95/CD95L axis and mediate initial immunopathology of COVID-19.
- Author
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Fraga-Silva TFC, Cipriano UG, Fumagalli MJ, Correa GF, Fuzo CA, Dos-Santos D, Mestriner FLAC, Becari C, Teixeira-Carvalho A, Coelho-Dos-Reis J, Menegueti MG, Figueiredo LTM, Cunha LD, Martins-Filho OA, Dias-Baruffi M, Auxiliadora-Martins M, Tostes RC, and Bonato VLD
- Abstract
Airway epithelial cells (AEC) infected with SARS-CoV-2 may drive the dysfunction of macrophages during COVID-19. We hypothesized that the direct interaction of AEC with macrophages mediated by CD95/CD95L or indirect interaction mediated by IL-6 signaling are key steps for the COVID-19 severe acute inflammation. The interaction of macrophages with apoptotic and infected AEC increased CD95 and CD163 expression, and induced macrophage death. Macrophages exposed to tracheal aspirate with high IL-6 levels from intubated patients with COVID-19 or to recombinant human IL-6 exhibited decreased HLA-DR expression, increased CD95 and CD163 expression and IL-1β production. IL-6 effects on macrophages were prevented by both CD95/CD95L antagonist and by IL-6 receptor antagonist and IL-6 or CD95 deficient mice showed significant reduction of acute pulmonary inflammation post-infection. Our findings show a non-canonical CD95L-CD95 pathway that simultaneously drives both macrophage activation and dysfunction and point to CD95/CD95L axis as therapeutic target., Competing Interests: The authors declare no competing interests., (© 2023 The Authors.)
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- 2023
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10. Cytokine storm in individuals with severe COVID-19 decreases endothelial cell antioxidant defense via downregulation of the Nrf2 transcriptional factor.
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Rodrigues D, Machado MR, Alves JV, Fraga-Silva TFC, Martins RB, Campos LCB, Francisco DF, Couto AES, Bonato VLD, Arruda E, Becari C, Auxiliadora-Martins M, Louzada-Júnior P, Costa RM, and Tostes RC
- Subjects
- Humans, NF-E2-Related Factor 2 metabolism, Down-Regulation, Cytokine Release Syndrome, Interleukin-6 metabolism, Cells, Cultured, SARS-CoV-2 metabolism, Oxidative Stress, Human Umbilical Vein Endothelial Cells metabolism, Reactive Oxygen Species metabolism, Cytokines metabolism, Antioxidants pharmacology, Antioxidants metabolism, COVID-19
- Abstract
The cytokine storm in SARS-CoV-2 infection contributes to the onset of inflammation and target-organ damage. The endothelium is a key player in COVID-19 pathophysiology and it is an important target for cytokines. Considering that cytokines trigger oxidative stress and negatively impact endothelial cell function, we sought to determine whether serum from individuals with severe COVID-19 decreases endothelial cells' main antioxidant defense, i.e., the antioxidant transcriptional factor Nrf2. Human umbilical vein endothelial cells (HUVECs) were incubated with serum from patients with severe COVID-19 at different time points and the effects on redox balance and Nrf2 activity were determined. Serum from individuals with COVID-19 increased oxidant species, as indicated by higher DHE (dihydroethydine) oxidation, increased protein carbonylation, and induced mitochondrial reactive oxygen species (ROS) generation and dysfunction. Serum from patients with COVID-19, but not serum from healthy individuals, induced cell death and diminished nitric oxide (NO) bioavailability. In parallel, Nrf2 nuclear accumulation and the expression of Nrf2-targeted genes were decreased in endothelial cells exposed to serum from individuals with COVID-19. In addition, these cells exhibited higher expression of Bach-1, a negative regulator of Nrf2 that competes for DNA binding. All events were prevented by tocilizumab, an IL-6 receptor blocker, indicating that IL-6 is key to the impairment of endothelial antioxidant defense. In conclusion, endothelial dysfunction related to SARS-CoV-2 infection is linked to decreased endothelial antioxidant defense via IL-6-dependent mechanisms. Pharmacological activation of Nrf2 may decrease endothelial cell damage in individuals with severe COVID-19. NEW & NOTEWORTHY We demonstrate that endothelial cell dysfunction in SARS-CoV-2-infected individuals is linked to decreased activity of the major antioxidant system regulator, the Nrf2 transcription factor. We provide evidence that this phenomenon relies on IL-6, an important cytokine involved in the pathophysiology of COVID-19. Our data support the view that Nrf2 activation is a potential therapeutical strategy to prevent oxidative stress and vascular inflammation in severe cases of COVID-19.
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- 2023
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11. Plasma Sphingomyelin Disturbances: Unveiling Its Dual Role as a Crucial Immunopathological Factor and a Severity Prognostic Biomarker in COVID-19.
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Toro DM, da Silva-Neto PV, de Carvalho JCS, Fuzo CA, Pérez MM, Pimentel VE, Fraga-Silva TFC, Oliveira CNS, Caruso GR, Vilela AFL, Nobre-Azevedo P, Defelippo-Felippe TV, Argolo JGM, Degiovani AM, Ostini FM, Feitosa MR, Parra RS, Vilar FC, Gaspar GG, Rocha JJRD, Feres O, Costa GP, Maruyama SRC, Russo EMS, Fernandes APM, Santos IKFM, Malheiro A, Sadikot RT, Bonato VLD, Cardoso CRB, Dias-Baruffi M, Trapé ÁA, Faccioli LH, Sorgi CA, and ImmunoCovid Consortium Group
- Subjects
- Humans, Prognosis, SARS-CoV-2 metabolism, Ceramides metabolism, Sphingolipids metabolism, Biomarkers, Sphingomyelins, COVID-19
- Abstract
SARS-CoV-2 infection triggers distinct patterns of disease development characterized by significant alterations in host regulatory responses. Severe cases exhibit profound lung inflammation and systemic repercussions. Remarkably, critically ill patients display a "lipid storm", influencing the inflammatory process and tissue damage. Sphingolipids (SLs) play pivotal roles in various cellular and tissue processes, including inflammation, metabolic disorders, and cancer. In this study, we employed high-resolution mass spectrometry to investigate SL metabolism in plasma samples obtained from control subjects ( n = 55), COVID-19 patients ( n = 204), and convalescent individuals ( n = 77). These data were correlated with inflammatory parameters associated with the clinical severity of COVID-19. Additionally, we utilized RNAseq analysis to examine the gene expression of enzymes involved in the SL pathway. Our analysis revealed the presence of thirty-eight SL species from seven families in the plasma of study participants. The most profound alterations in the SL species profile were observed in patients with severe disease. Notably, a predominant sphingomyelin (SM d18:1) species emerged as a potential biomarker for COVID-19 severity, showing decreased levels in the plasma of convalescent individuals. Elevated SM levels were positively correlated with age, hospitalization duration, clinical score, and neutrophil count, as well as the production of IL-6 and IL-8. Intriguingly, we identified a putative protective effect against disease severity mediated by SM (d18:1/24:0), while ceramide (Cer) species (d18:1/24:1) and (d18:1/24:0)were associated with increased risk. Moreover, we observed the enhanced expression of key enzymes involved in the SL pathway in blood cells from severe COVID-19 patients, suggesting a primary flow towards Cer generation in tandem with SM synthesis. These findings underscore the potential of SM as a prognostic biomarker for COVID-19 and highlight promising pharmacological targets. By targeting sphingolipid pathways, novel therapeutic strategies may emerge to mitigate the severity of COVID-19 and improve patient outcomes.
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- 2023
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12. Pioneering in vitro characterization of macrophage response induced by scorpion venoms from the Brazilian Amazon.
- Author
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Reis MB, Martins JG, Bordon KCF, de Campos Fraga-Silva TF, de Lima Procópio RE, de Almeida BRR, Bonato VLD, and Arantes EC
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- Humans, Mice, Animals, Brazil, Scorpions, Macrophages, Scorpion Venoms toxicity, Scorpion Stings
- Abstract
There are several scorpion species of medical relevance around the world. Some of them are well characterized by their toxins and clinical outcomes. Brazilian Amazon has a great amount of these arthropods that have an impact in the scorpionism events specifically in this region of Brazil. Recently, several studies pointed out the immune system activation during scorpion envenouming as an important facet of scorpionism, inducing a sepsis-like state that culminates in clinical severity and death. In this work, we characterized the macrophage response of three species of clinical relevance in Brazilian Amazon: Tityus silvestris, T. metuendus and T. obscurus and one specie with no toxic effects to humans, Brotheas amazonicus. All the four species analyzed were able to induce pro- and anti-inflammatory cytokine production in a J774.1 murine macrophage model. This activation was dependent on TLR2/TLR4/MyD88 activation and abolished by TLRs antagonists. These results suggest that the venoms of the four species analyzed were able to induce macrophage response in agreement to the well-established immune activation by T. serrulatus venom. Our findings provide new insights into the clinical repercussions of scorpionism of uncharacterized species and point to new biotechnological applications of these venoms and possible supportive therapies in scorpionism., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)
- Published
- 2023
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13. The turning point of COVID-19 severity is associated with a unique circulating neutrophil gene signature.
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Fuzo CA, Fraga-Silva TFC, Maruyama SR, Bastos VAF, Rogerio LA, Takamiya NT, da Silva-Neto PV, Pimentel VE, Toro DM, Pérez MM, de Carvalho JCS, Carmona-Garcia I, Oliveira CNS, Degiovani AM, Ostini FM, Constant LF, de Amorim AP, Vilar FC, Feitosa MR, Parra RS, da Rocha JJR, Feres O, Gaspar GG, Viana AL, Fernandes APM, Santos IKFM, Russo EMS, Cardoso CRB, Sorgi CA, Faccioli LH, Bonato VLD, and Dias-Baruffi M
- Subjects
- Humans, Neutrophils, Transcriptome, Biomarkers, COVID-19 genetics
- Abstract
COVID-19 has a broad spectrum of clinical manifestations associated with the host immune response heterogeneity. Despite the advances in COVID-19 research, it is still crucial to seek a panel of molecular markers that enable accurate stratification of COVID-19 patients. Here, we performed a study that combined analysis of blood transcriptome, demographic data, clinical aspects and laboratory findings from 66 participants classified into different degrees of COVID-19 severity and healthy subjects. We identified a perturbation in blood-leukocyte transcriptional profile associated with COVID-19 aggravation, which was mainly related to processes that disfavoured lymphocyte activation and favoured neutrophil activation. This transcriptional profile stratified patients according to COVID-19 severity. Hence, it enabled identification of a turning point in transcriptional dynamics that distinguished disease outcomes and non-hospitalized from hospitalized moderate patients. Central genes of this unique neutrophil signature were S100A9, ANXA3, CEACAM6, VNN1, OLFM4, IL1R2, TCN1 and CD177. Our study indicates the molecular changes that are linked with the differing clinical aspects presented by humans when suffering from COVID-19, which involve neutrophil activation., (© 2023 John Wiley & Sons Ltd.)
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- 2023
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14. IL-22 Is Deleterious along with IL-17 in Allergic Asthma but Is Not Detrimental in the Comorbidity Asthma and Acute Pneumonia.
- Author
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Goulart A, Boko MMM, Martins NS, Gembre AF, de Oliveira RS, Palma-Albornoz SP, Bertolini T, Ribolla PEM, Ramalho LNZ, Fraga-Silva TFC, and Bonato VLD
- Subjects
- Mice, Animals, Interleukin-17 genetics, Lung pathology, Eosinophils, Allergens, Comorbidity, Ovalbumin, Disease Models, Animal, Bronchoalveolar Lavage Fluid, Mice, Inbred BALB C, Asthma pathology, Pneumonia pathology
- Abstract
There is evidence that IL-22 and IL-17 participate in the pathogenesis of allergic asthma. To investigate the role of IL-22, we used IL-22 deficient mice (IL-22 KO) sensitized and challenged with ovalbumin (OVA) and compared with wild type (WT) animals exposed to OVA. IL-22 KO animals exposed to OVA showed a decreased number and frequency of eosinophils, IL-5 and IL-13 in the airways, reduced mucus production and pulmonary inflammation. In addition, IL-22 KO animals exhibited a decreased percentage and number of lung CD11c
+ CD11b+ cells and increased apoptosis of eosinophils. Th17 cell transfer generated from IL-22 KO to animals previously sensitized and challenged with OVA caused a reduction in eosinophil frequency and number in the airways compared to animals transferred with Th17 cells generated from WT mice. Therefore, IL-22 is deleterious with concomitant secretion of IL-17. Our findings show a pro-inflammatory role for IL-22, confirmed in a model of allergen-free and allergen-specific immunotherapy. Moreover, during the comorbidity asthma and pneumonia that induces neutrophil inflammation, IL-22 was not detrimental. Our results show that targeting IL-22 would negatively affect the survival of eosinophils, reduce the expansion or migration of CD11c+ CD11b+ cells, and negatively regulate allergic asthma.- Published
- 2023
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15. The Interplay among Glucocorticoid Therapy, Platelet-Activating Factor and Endocannabinoid Release Influences the Inflammatory Response to COVID-19.
- Author
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de Carvalho JCS, da Silva-Neto PV, Toro DM, Fuzo CA, Nardini V, Pimentel VE, Pérez MM, Fraga-Silva TFC, Oliveira CNS, Degiovani AM, Ostini FM, Feitosa MR, Parra RS, da Rocha JJR, Feres O, Vilar FC, Gaspar GG, Santos IKFM, Fernandes APM, Maruyama SR, Russo EMS, Bonato VLD, Cardoso CRB, Dias-Baruffi M, Faccioli LH, Sorgi CA, and On Behalf Of The ImmunoCovid Study Group
- Subjects
- Humans, Cross-Sectional Studies, Endocannabinoids, Glucocorticoids therapeutic use, Platelet Activating Factor, COVID-19
- Abstract
COVID-19 is associated with a dysregulated immune response. Currently, several medicines are licensed for the treatment of this disease. Due to their significant role in inhibiting pro-inflammatory cytokines and lipid mediators, glucocorticoids (GCs) have attracted a great deal of attention. Similarly, the endocannabinoid (eCB) system regulates various physiological processes including the immunological response. Additionally, during inflammatory and thrombotic processes, phospholipids from cell membranes are cleaved to produce platelet-activating factor (PAF), another lipid mediator. Nonetheless, the effect of GCs on this lipid pathway during COVID-19 therapy is still unknown. This is a cross-sectional study involving COVID-19 patients ( n = 200) and healthy controls ( n = 35). Target tandem mass spectrometry of plasma lipid mediators demonstrated that COVID-19 severity affected eCBs and PAF synthesis. This increased synthesis of eCB was adversely linked with systemic inflammatory markers IL-6 and sTREM-1 levels and neutrophil counts. The use of GCs altered these lipid pathways by reducing PAF and increasing 2-AG production. Corroborating this, transcriptome analysis of GC-treated patients blood leukocytes showed differential modulation of monoacylglycerol lipase and phospholipase A2 gene expression. Altogether, these findings offer a breakthrough in our understanding of COVID-19 pathophysiology, indicating that GCs may promote additional protective pharmacological effects by influencing the eCB and PAF pathways involved in the disease course.
- Published
- 2023
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16. Xenogeneic mesenchymal stem cell biocurative improves skin wounds healing in diabetic mice by increasing mast cells and the regenerative profile.
- Author
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Manso GMDC, Elias-Oliveira J, Guimarães JB, Pereira ÍS, Rodrigues VF, Burger B, Fantacini DMC, de Souza LEB, Rodrigues HG, Bonato VLD, Silva JS, Ramos SG, Tostes RC, Manfiolli AO, Caliari-Oliveira C, and Carlos D
- Abstract
Introduction: Diabetes mellitus (DM) is a chronic disease and a major cause of mortality and morbidity worldwide. The hyperglycemia caused by DM induces micro and macrovascular complications that lead, among other consequences, to chronic wounds and amputations. Cell therapy and tissue engineering constitute recent therapeutic alternatives to improve wound healing in diabetic patients. The current study aimed to analyze the effectiveness of biocuratives containing human mesenchymal stem cells (MSCs) associated with a hydrogel matrix in the wound healing process and related inflammatory cell profile in diabetic mice., Methods: Biocuratives containing MSCs were constructed by 3D bioprinting, and applied to skin wounds on the back of streptozotocin (STZ)-induced type 1 diabetic (T1D) mice. The healing process, after the application of biocuratives with or without MSCs was histologically analyzed. In parallel, genes related to growth factors, mast cells (MC), M1 and M2 macrophage profiles were evaluated by RT-PCR. Macrophages were characterized by flow cytometry, and MC by toluidine blue staining and flow cytometry., Results: Mice with T1D exhibited fewer skin MC and delayed wound healing when compared to the non-diabetic group. Treatment with the biocuratives containing MSCs accelerated wound healing and improved skin collagen deposition in diabetic mice. Increased TGF-β gene expression and M2 macrophage-related markers were also detected in skin of diabetic mice that received MSCs-containing biocuratives. Finally, MSCs upregulated IL-33 gene expression and augmented the number of MC in the skin of diabetic mice., Conclusion: These results reveal the therapeutic potential of biocuratives containing MSCs in the healing of skin wounds in diabetic mice, providing a scientific base for future treatments in diabetic patients., Competing Interests: CCO and AOM are partners is In Situ Cell Therapy and are named as inventors of a provisional patent directed at this manuscript, which is solely owned by In Situ Cell Therapy., (© 2022 The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V.)
- Published
- 2023
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17. The effects of female sexual hormones on the endothelial glycocalyx.
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Potje SR, Martins NS, Benatti MN, Rodrigues D, Bonato VLD, and Tostes RC
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- Pregnancy, Humans, Female, Endothelial Cells metabolism, Vasodilation, Estrogens metabolism, Estrogens pharmacology, Progesterone metabolism, Progesterone pharmacology, Glycocalyx metabolism
- Abstract
The glycocalyx is a layer composed of carbohydrate side chains bound to core proteins that lines the vascular endothelium. The integrity of the glycocalyx is essential for endothelial cells' performance and vascular homeostasis. The neuroendocrine and immune systems influence the composition, maintenance, activity and degradation of the endothelial glycocalyx. The female organism has unique characteristics, and estrogen and progesterone, the main female hormones are essential to the development and physiology of the reproductive system and to the ability to develop a fetus. Female sex hormones also exert a wide variety of effects on other organs, including the vascular endothelium. They upregulate nitric oxide synthase expression and activity, decrease oxidative stress, increase vasodilation, and protect from vascular injury. This review will discuss how female hormones and pregnancy, which prompts to high levels of estrogen and progesterone, modulate the endothelial glycocalyx. Diseases prevalent in women that alter the glycocalyx, and therapeutic forms to prevent glycocalyx degradation and potential treatments that can reconstitute its structure and function will also be discussed., Competing Interests: Conflict of interest The authors declare that this manuscript was written in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Elsevier Inc. All rights reserved.)
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- 2023
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18. Celastrol: A lead compound that inhibits SARS-CoV-2 replication, the activity of viral and human cysteine proteases, and virus-induced IL-6 secretion.
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Fuzo CA, Martins RB, Fraga-Silva TFC, Amstalden MK, Canassa De Leo T, Souza JP, Lima TM, Faccioli LH, Okamoto DN, Juliano MA, França SC, Juliano L, Bonato VLD, Arruda E, and Dias-Baruffi M
- Subjects
- Humans, Interleukin-6, Molecular Docking Simulation, Protease Inhibitors pharmacology, SARS-CoV-2 drug effects, Viral Nonstructural Proteins genetics, Viral Nonstructural Proteins metabolism, Antiviral Agents pharmacology, Coronavirus 3C Proteases antagonists & inhibitors, Pentacyclic Triterpenes pharmacology, COVID-19 Drug Treatment
- Abstract
The global emergence of coronavirus disease 2019 (COVID-19) has caused substantial human casualties. Clinical manifestations of this disease vary from asymptomatic to lethal, and the symptomatic form can be associated with cytokine storm and hyperinflammation. In face of the urgent demand for effective drugs to treat COVID-19, we have searched for candidate compounds using in silico approach followed by experimental validation. Here we identified celastrol, a pentacyclic triterpene isolated from Tripterygium wilfordii Hook F, as one of the best compounds out of 39 drug candidates. Celastrol reverted the gene expression signature from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected cells and irreversibly inhibited the recombinant forms of the viral and human cysteine proteases involved in virus invasion, such as M
pro (main protease), PLpro (papain-like protease), and recombinant human cathepsin L. Celastrol suppressed SARS-CoV-2 replication in human and monkey cell lines and decreased interleukin-6 (IL-6) secretion in the SARS-CoV-2-infected human cell line. Celastrol acted in a concentration-dependent manner, with undetectable signs of cytotoxicity, and inhibited in vitro replication of the parental and SARS-CoV-2 variant. Therefore, celastrol is a promising lead compound to develop new drug candidates to face COVID-19 due to its ability to suppress SARS-CoV-2 replication and IL-6 production in infected cells., (© 2022 Wiley Periodicals LLC.)- Published
- 2022
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19. Mast cell-T cell axis alters development of colitis-dependent and colitis-independent colorectal tumours: potential for therapeutically targeting via mast cell inhibition.
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Sakita JY, Elias-Oliveira J, Carlos D, de Souza Santos E, Almeida LY, Malta TM, Brunaldi MO, Albuquerque S, Araújo Silva CL, Andrade MV, Bonato VLD, Garcia SB, Cunha FQ, Cebinelli GCM, Martins RB, Matthews J, Colli L, Martin FL, Uyemura SA, and Kannen V
- Subjects
- Animals, Fluorouracil, Humans, Mast Cells, Mice, Colitis, Colorectal Neoplasms
- Abstract
Background: Colorectal cancer (CRC) has a high mortality rate and can develop in either colitis-dependent (colitis-associated (CA)-CRC) or colitis-independent (sporadic (s)CRC) manner. There has been a significant debate about whether mast cells (MCs) promote or inhibit the development of CRC. Herein we investigated MC activity throughout the multistepped development of CRC in both human patients and animal models., Methods: We analyzed human patient matched samples of healthy colon vs CRC tissue alongside conducting a The Cancer Genome Atlas-based immunogenomic analysis and multiple experiments employing genetically engineered mouse (GEM) models., Results: Analyzing human CRC samples revealed that MCs can be active or inactive in this disease. An activated MC population decreased the number of tumor-residing CD8 T cells. In mice, MC deficiency decreased the development of CA-CRC lesions, while it increased the density of tumor-based CD8 infiltration. Furthermore, co-culture experiments revealed that tumor-primed MCs promote apoptosis in CRC cells. In MC-deficient mice, we found that MCs inhibited the development of sCRC lesions. Further exploration of this with several GEM models confirmed that different immune responses alter and are altered by MC activity, which directly alters colon tumorigenesis. Since rescuing MC activity with bone marrow transplantation in MC-deficient mice or pharmacologically inhibiting MC effects impacts the development of sCRC lesions, we explored its therapeutic potential against CRC. MC activity promoted CRC cell engraftment by inhibiting CD8+ cell infiltration in tumors, pharmacologically blocking it inhibits the ability of allograft tumors to develop. This therapeutic strategy potentiated the cytotoxic activity of fluorouracil chemotherapy., Conclusion: Therefore, we suggest that MCs have a dual role throughout CRC development and are potential druggable targets against this disease., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)
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- 2022
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20. Asthma-associated bacterial infections: Are they protective or deleterious?
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Fraga-Silva TFC, Boko MMM, Martins NS, Cetlin AA, Russo M, Vianna EO, and Bonato VLD
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Eosinophilic, noneosinophilic, or mixed granulocytic inflammations are the hallmarks of asthma heterogeneity. Depending on the priming of lung immune and structural cells, subjects with asthma might generate immune responses that are T
H 2-prone or TH 17-prone immune response. Bacterial infections caused by Haemophilus , Moraxella , or Streptococcus spp. induce the secretion of IL-17, which in turn recruit neutrophils into the airways. Clinical studies and experimental models of asthma indicated that neutrophil infiltration induces a specific phenotype of asthma, characterized by an impaired response to corticosteroid treatment. The understanding of pathways that regulate the TH 17-neutrophils axis is critical to delineate and develop host-directed therapies that might control asthma and its exacerbation episodes that course with infectious comorbidities. In this review, we outline clinical and experimental studies on the role of airway epithelial cells, S100A9, and high mobility group box 1, which act in concert with the IL-17-neutrophil axis activated by bacterial infections, and are related with asthma that is difficult to treat. Furthermore, we report critically our view in the light of these findings in an attempt to stimulate further investigations and development of immunotherapies for the control of severe asthma., (© 2022 Published by Elsevier Inc. on behalf of the American Academy of Allergy, Asthma & Immunology.)- Published
- 2022
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21. The Severity of COVID-19 Affects the Plasma Soluble Levels of the Immune Checkpoint HLA-G Molecule.
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Cordeiro JFC, Fernandes TM, Toro DM, da Silva-Neto PV, Pimentel VE, Pérez MM, de Carvalho JCS, Fraga-Silva TFC, Oliveira CNS, Argolo JGM, Degiovani AM, Ostini FM, Puginna EF, da Silva JS, Santos IKFM, Bonato VLD, Cardoso CRB, Dias-Baruffi M, Faccioli LH, Donadi EA, Sorgi CA, Fernandes APM, and On Behalf Of The Immunocovid Study Group
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- Histocompatibility Antigens Class I, Humans, Immune Checkpoint Proteins, Plasma, SARS-CoV-2, COVID-19, HLA-G Antigens
- Abstract
The non-classical histocompatibility antigen G (HLA-G) is an immune checkpoint molecule that has been implicated in viral disorders. We evaluated the plasma soluble HLA-G (sHLA-G) in 239 individuals, arranged in COVID-19 patients ( n = 189) followed up at home or in a hospital, and in healthy controls ( n = 50). Increased levels of sHLA-G were observed in COVID-19 patients irrespective of the facility care, gender, age, and the presence of comorbidities. Compared with controls, the sHLA-G levels increased as far as disease severity progressed; however, the levels decreased in critically ill patients, suggesting an immune exhaustion phenomenon. Notably, sHLA-G exhibited a positive correlation with other mediators currently observed in the acute phase of the disease, including IL-6, IL-8 and IL-10. Although sHLA-G levels may be associated with an acute biomarker of COVID-19, the increased levels alone were not associated with disease severity or mortality due to COVID-19. Whether the SARS-CoV-2 per se or the innate/adaptive immune response against the virus is responsible for the increased levels of sHLA-G are questions that need to be further addressed.
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- 2022
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22. RIP2 Contributes to Expanded CD4 + T Cell IFN-γ Production during Efferocytosis of Streptococcus pneumoniae -Infected Apoptotic Cells.
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Niño-Castaño VE, Penteado LA, Silva-Pereira L, Bazzano JMR, Orlando AB, Salina ACG, Dejani NN, Bonato VLD, Serezani CH, and Medeiros AI
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- Animals, Apoptosis, Interferon-gamma metabolism, Mice, Mice, Inbred C57BL, Signal Transduction, Th1 Cells, CD4-Positive T-Lymphocytes, Interferon-gamma biosynthesis, Streptococcus pneumoniae, Toll-Like Receptor 2 metabolism, Ubiquitin-Protein Ligases metabolism
- Abstract
Apoptotic cell clearance by professional and nonprofessional phagocytes in the process of efferocytosis is critical to preserve tissue homeostasis. Uptake of apoptotic cells by dendritic cells generates regulatory T cells and induces immunologic tolerance against self-antigens. In contrast, ingestion of infected apoptotic cells promotes activation of TLR4/MyD88-dependent bone marrow-derived dendritic cells (BMDCs) and triggers Th17 cell differentiation. In this study, we evaluated the impact of Streptococcus pneumoniae -infected apoptotic cell efferocytosis by BMDCs derived from C57BL/6 mice on differentiation and expansion of CD4
+ T cell subsets, as well as the role of TLR2/4 and receptor-interacting protein 2 (RIP2) receptors in recognizing intracellular pathogens during efferocytosis. We demonstrated that BMDC-mediated efferocytosis of S. pneumoniae -infected apoptotic cells induced Th1 cell differentiation and expansion. Although TLR2/4 and RIP2 deficiency in BMDCs did not affect Th1 cell differentiation during efferocytosis, the absence of RIP2 decreased IFN-γ production by CD4 T cells during the expansion phase. These findings suggest that RIP2-mediated IL-1β production during efferocytosis of S. pneumoniae -infected apoptotic cells partially supports a Th1-mediated IFN-γ production microenvironment., (Copyright © 2022 The Authors.)- Published
- 2022
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23. Acetylcholine, Fatty Acids, and Lipid Mediators Are Linked to COVID-19 Severity.
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Pérez MM, Pimentel VE, Fuzo CA, da Silva-Neto PV, Toro DM, Fraga-Silva TFC, Gardinassi LG, Oliveira CNS, Souza COS, Torre-Neto NT, de Carvalho JCS, De Leo TC, Nardini V, Feitosa MR, Parra RS, da Rocha JJR, Feres O, Vilar FC, Gaspar GG, Constant LF, Ostini FM, Degiovani AM, Amorim AP, Viana AL, Fernandes APM, Maruyama SR, Russo EMS, Santos IKFM, Bonato VLD, Cardoso CRB, Sorgi CA, Dias-Baruffi M, and Faccioli LH
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- Arachidonic Acid, Arachidonic Acids pharmacology, Fatty Acids, Glucocorticoids, Humans, SARS-CoV-2, Acetylcholine, COVID-19
- Abstract
Lipid and cholinergic mediators are inflammatory regulators, but their role in the immunopathology of COVID-19 is still unclear. Here, we used human blood and tracheal aspirate (TA) to investigate whether acetylcholine (Ach), fatty acids (FAs), and their derived lipid mediators (LMs) are associated with COVID-19 severity. First, we analyzed the perturbation profile induced by SARS-CoV-2 infection in the transcriptional profile of genes related to the ACh and FA/LM pathways. Blood and TA were used for metabolomic and lipidomic analyses and for quantification of leukocytes, cytokines, and ACh. Differential expression and coexpression gene network data revealed a unique transcriptional profile associated with ACh and FA/LM production, release, and cellular signaling. Transcriptomic data were corroborated by laboratory findings: SARS-CoV-2 infection increased plasma and TA levels of arachidonic acid, 5-hydroxy - 6 E, 8 Z, 11 Z, 14 Z -eicosatetraenoic acid, 11-hydroxy-5 Z, 8 Z, 12 E, 14 Z- eicosatetraenoic acid, and ACh. TA samples also exhibited high levels of PGE
2 , thromboxane B2 , 12-oxo - 5 Z, 8 Z, 10 E, 14 Z- eicosatetraenoic acid, and 6- trans -leukotriene B4 Bioinformatics and experimental approaches demonstrated robust correlation between transcriptional profile in Ach and FA/LM pathways and parameters of severe COVID-19. As expected, the increased neutrophil-to-lymphocyte ratio, neutrophil counts, and cytokine levels (IL-6, IL-10, IL-1β, and IL-8) correlated with worse clinical scores. Glucocorticoids protected severe and critical patients and correlated with reduced Ach levels in plasma and TA samples. We demonstrated that pulmonary and systemic hyperinflammation in severe COVID-19 are associated with high levels of Ach and FA/LM. Glucocorticoids favored the survival of patients with severe/critical disease, and this effect was associated with a reduction in ACh levels.- Published
- 2022
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24. Efferocytosis of SARS-CoV-2-infected dying cells impairs macrophage anti-inflammatory functions and clearance of apoptotic cells.
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Salina ACG, Dos-Santos D, Rodrigues TS, Fortes-Rocha M, Freitas-Filho EG, Alzamora-Terrel DL, Castro IMS, Fraga da Silva TFC, de Lima MHF, Nascimento DC, Silva CM, Toller-Kawahisa JE, Becerra A, Oliveira S, Caetité DB, Almeida L, Ishimoto AY, Lima TM, Martins RB, Veras F, do Amaral NB, Giannini MC, Bonjorno LP, Lopes MIF, Benatti MN, Batah SS, Santana RC, Vilar FC, Martins MA, Assad RL, de Almeida SCL, de Oliveira FR, Arruda Neto E, Cunha TM, Alves-Filho JC, Bonato VLD, Cunha FQ, Fabro AT, Nakaya HI, Zamboni DS, Louzada-Junior P, Oliveira RDR, and Cunha LD
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- Anti-Inflammatory Agents pharmacology, Apoptosis, Humans, Macrophages metabolism, Phagocytosis, COVID-19, SARS-CoV-2
- Abstract
COVID-19 is a disease of dysfunctional immune responses, but the mechanisms triggering immunopathogenesis are not established. The functional plasticity of macrophages allows this cell type to promote pathogen elimination and inflammation or suppress inflammation and promote tissue remodeling and injury repair. During an infection, the clearance of dead and dying cells, a process named efferocytosis, can modulate the interplay between these contrasting functions. Here, we show that engulfment of SARS-CoV-2-infected apoptotic cells exacerbates inflammatory cytokine production, inhibits the expression of efferocytic receptors, and impairs continual efferocytosis by macrophages. We also provide evidence supporting that lung monocytes and macrophages from severe COVID-19 patients have compromised efferocytic capacity. Our findings reveal that dysfunctional efferocytosis of SARS-CoV-2-infected cell corpses suppresses macrophage anti-inflammation and efficient tissue repair programs and provides mechanistic insights for the excessive production of pro-inflammatory cytokines and accumulation of tissue damage associated with COVID-19 immunopathogenesis., Competing Interests: AS, Dd, TR, MF, EF, DA, IC, TF, Md, DN, CS, JT, AB, SO, DC, LA, AI, TL, RM, FV, Nd, MG, LB, ML, MB, SB, RS, FV, MM, RA, Sd, Fd, EA, TC, JA, VB, FC, AF, HN, DZ, PL, RO, LC No competing interests declared, (© 2022, Salina, dos-Santos, Rodrigues et al.)
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- 2022
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25. Timeline Kinetics of Systemic and Airway Immune Mediator Storm for Comprehensive Analysis of Disease Outcome in Critically Ill COVID-19 Patients.
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Gonçalves JJ, da Mata CPSM, Lourenço AA, Ribeiro ÁL, Ferreira GM, Fraga-Silva TFC, de Souza FM, Almeida VES, Batista IA, D Avila-Mesquita C, Couto AES, Campos LCB, Paim AAO, Ferreira LL, de Melo Oliveira P, de Almeida Teixeira L, Priscila de Almeida Marques D, Retes de Moraes H, Pereira SH, Brito-de-Sousa JP, Campi-Azevedo AC, Peruhype-Magalhães V, Araújo MSS, Teixeira-Carvalho A, da Fonseca FG, Bonato VLD, Becari C, Ferro D, Menegueti MG, Mazzoni AAS, Auxiliadora-Martins M, Coelho-Dos-Reis JG, and Martins-Filho OA
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- Critical Illness, Cytokines metabolism, Humans, Kinetics, SARS-CoV-2, COVID-19
- Abstract
In the present study, the levels of serum and airway soluble chemokines, pro-inflammatory/regulatory cytokines, and growth factors were quantified in critically ill COVID-19 patients (total n=286) at distinct time points (D0, D2-6, D7, D8-13 and D>14-36) upon Intensive Care Unit (ICU) admission. Augmented levels of soluble mediators were observed in serum from COVID-19 patients who progress to death. An opposite profile was observed in tracheal aspirate samples, indicating that systemic and airway microenvironment diverge in their inflammatory milieu. While a bimodal distribution was observed in the serum samples, a unimodal peak around D7 was found for most soluble mediators in tracheal aspirate samples. Systems biology tools further demonstrated that COVID-19 display distinct eccentric soluble mediator networks as compared to controls, with opposite profiles in serum and tracheal aspirates. Regardless the systemic-compartmentalized microenvironment, networks from patients progressing to death were linked to a pro-inflammatory/growth factor-rich, highly integrated center. Conversely, patients evolving to discharge exhibited networks of weak central architecture, with lower number of neighborhood connections and clusters of pro-inflammatory and regulatory cytokines. All in all, this investigation with robust sample size landed a comprehensive snapshot of the systemic and local divergencies composed of distinct immune responses driven by SARS-CoV-2 early on severe COVID-19., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Gonçalves, da Mata, Lourenço, Ribeiro, Ferreira, Fraga-Silva, de Souza, Almeida, Batista, D`Avila-Mesquita, Couto, Campos, Paim, Ferreira, de Melo Oliveira, de Almeida Teixeira, Priscila de Almeida Marques, Retes de Moraes, Pereira, Brito-de-Sousa, Campi-Azevedo, Peruhype-Magalhães, Araújo, Teixeira-Carvalho, da Fonseca, Bonato, Becari, Ferro, Menegueti, Mazzoni, Auxiliadora-Martins, Coelho-dos-Reis and Martins-Filho.)
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- 2022
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26. Matrix Metalloproteinases on Severe COVID-19 Lung Disease Pathogenesis: Cooperative Actions of MMP-8/MMP-2 Axis on Immune Response through HLA-G Shedding and Oxidative Stress.
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da Silva-Neto PV, do Valle VB, Fuzo CA, Fernandes TM, Toro DM, Fraga-Silva TFC, Basile PA, de Carvalho JCS, Pimentel VE, Pérez MM, Oliveira CNS, Rodrigues LC, Bastos VAF, Tella SOC, Martins RB, Degiovani AM, Ostini FM, Feitosa MR, Parra RS, Vilar FC, Gaspar GG, Rocha JJRD, Feres O, Arruda E, Maruyama SR, Russo EMS, Viana AL, Santos IKFM, Bonato VLD, Cardoso CRB, Tanus-Santos JE, Donadi EA, Faccioli LH, Dias-Baruffi M, Fernandes APM, Gerlach RF, Sorgi CA, and On Behalf Of The Immunocovid Study Group
- Subjects
- HLA-G Antigens, Humans, Immunity, Matrix Metalloproteinase 8 metabolism, Oxidative Stress, SARS-CoV-2, COVID-19, Matrix Metalloproteinase 2 metabolism
- Abstract
Patients with COVID-19 predominantly have a respiratory tract infection and acute lung failure is the most severe complication. While the molecular basis of SARS-CoV-2 immunopathology is still unknown, it is well established that lung infection is associated with hyper-inflammation and tissue damage. Matrix metalloproteinases (MMPs) contribute to tissue destruction in many pathological situations, and the activity of MMPs in the lung leads to the release of bioactive mediators with inflammatory properties. We sought to characterize a scenario in which MMPs could influence the lung pathogenesis of COVID-19. Although we observed high diversity of MMPs in lung tissue from COVID-19 patients by proteomics, we specified the expression and enzyme activity of MMP-2 in tracheal-aspirate fluid (TAF) samples from intubated COVID-19 and non-COVID-19 patients. Moreover, the expression of MMP-8 was positively correlated with MMP-2 levels and possible shedding of the immunosuppression mediator sHLA-G and sTREM-1. Together, overexpression of the MMP-2/MMP-8 axis, in addition to neutrophil infiltration and products, such as reactive oxygen species (ROS), increased lipid peroxidation that could promote intensive destruction of lung tissue in severe COVID-19. Thus, the inhibition of MMPs can be a novel target and promising treatment strategy in severe COVID-19.
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- 2022
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27. Systemic Infection by Non- albicans Candida Species Affects the Development of a Murine Model of Multiple Sclerosis.
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Fraga-Silva TFC, Munhoz-Alves N, Mimura LAN, Oliveira LRC, Figueiredo-Godoi LMA, Garcia MT, Oliveira ES, Ishikawa LLW, Zorzella-Pezavento SFG, Bonato VLD, Junqueira JC, Bagagli E, and Sartori A
- Abstract
Candidiasis may affect the central nervous system (CNS), and although Candida albicans is predominant, non- albicans Candida species can also be associated with CNS infections. Some studies have suggested that Candida infections could increase the odds of multiple sclerosis (MS) development. In this context, we investigated whether systemic infection by non- albicans Candida species would affect, clinically or immunologically, the severity of experimental autoimmune encephalomyelitis (EAE), which is an animal model used to study MS. For this, a strain of C. glabrata , C. krusei , and C. parapsilosis was selected and characterized using different in vitro and in vivo models. In these analysis, all the strains exhibited the ability to form biofilms, produce proteolytic enzymes, and cause systemic infections in Galleria mellonella , with C. glabrata being the most virulent species. Next, C57BL/6 mice were infected with strains of C. glabrata , C. krusei , or C. parapsilosis , and 3 days later were immunized with myelin oligodendrocyte glycoprotein to develop EAE. Mice from EAE groups previously infected with C. glabrata and C. krusei developed more severe and more prevalent paralysis, while mice from the EAE group infected with C. parapsilosis developed a disease comparable to non-infected EAE mice. Disease aggravation by C. glabrata and C. krusei strains was concomitant to increased IL-17 and IFN-γ production by splenic cells stimulated with fungi-derived antigens and with increased percentage of T lymphocytes and myeloid cells in the CNS. Analysis of interaction with BV-2 microglial cell line also revealed differences among these strains, in which C. krusei was the strongest activator of microglia concerning the expression of MHC II and CD40 and pro-inflammatory cytokine production. Altogether, these results indicated that the three non- albicans Candida strains were similarly able to reach the CNS but distinct in terms of their effect over EAE development. Whereas C. glabrata and C. Krusei aggravated the development of EAE, C. parapsilosis did not affect its severity. Disease worsening was partially associated to virulence factors in C. glabrata and to a strong activation of microglia in C. krusei infection. In conclusion, systemic infections by non- albicans Candida strains exerted influence on the experimental autoimmune encephalomyelitis in both immunological and clinical aspects, emphasizing their possible relevance in MS development.
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- 2022
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28. Mitochondrial DNA and TLR9 activation contribute to SARS-CoV-2-induced endothelial cell damage.
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Costa TJ, Potje SR, Fraga-Silva TFC, da Silva-Neto JA, Barros PR, Rodrigues D, Machado MR, Martins RB, Santos-Eichler RA, Benatti MN, de Sá KSG, Almado CEL, Castro ÍA, Pontelli MC, Serra L, Carneiro FS, Becari C, Louzada-Junior P, Oliveira RDR, Zamboni DS, Arruda E, Auxiliadora-Martins M, Giachini FRC, Bonato VLD, Zachara NE, Bomfim GF, and Tostes RC
- Subjects
- Animals, Endothelial Cells metabolism, Humans, Mice, Mitochondria metabolism, SARS-CoV-2, Toll-Like Receptor 9 genetics, Toll-Like Receptor 9 metabolism, COVID-19, DNA, Mitochondrial genetics, DNA, Mitochondrial metabolism
- Abstract
Background and Purpose: Mitochondria play a central role in the host response to viral infection and immunity, being key to antiviral signaling and exacerbating inflammatory processes. Mitochondria and Toll-like receptor (TLR) have been suggested as potential targets in SARS-CoV-2 infection. However, the involvement of TLR9 in SARS-Cov-2-induced endothelial dysfunction and potential contribution to cardiovascular complications in COVID-19 have not been demonstrated. This study determined whether infection of endothelial cells by SARS-CoV-2 affects mitochondrial function and induces mitochondrial DNA (mtDNA) release. We also questioned whether TLR9 signaling mediates the inflammatory responses induced by SARS-CoV-2 in endothelial cells., Experimental Approach: Human umbilical vein endothelial cells (HUVECs) were infected by SARS-CoV-2 and immunofluorescence was used to confirm the infection. Mitochondrial function was analyzed by specific probes and mtDNA levels by real-time polymerase chain reaction (RT-PCR). Inflammatory markers were measured by ELISA, protein expression by western blot, intracellular calcium (Ca
2+ ) by FLUOR-4, and vascular reactivity with a myography., Key Results: SARS-CoV-2 infected HUVECs, which express ACE2 and TMPRSS2 proteins, and promoted mitochondrial dysfunction, i.e. it increased mitochondria-derived superoxide anion, mitochondrial membrane potential, and mtDNA release, leading to activation of TLR9 and NF-kB, and release of cytokines. SARS-CoV-2 also decreased nitric oxide synthase (eNOS) expression and inhibited Ca2+ responses in endothelial cells. TLR9 blockade reduced SARS-CoV-2-induced IL-6 release and prevented decreased eNOS expression. mtDNA increased vascular reactivity to endothelin-1 (ET-1) in arteries from wild type, but not TLR9 knockout mice. These events were recapitulated in serum samples from COVID-19 patients, that exhibited increased levels of mtDNA compared to sex- and age-matched healthy subjects and patients with comorbidities., Conclusion and Applications: SARS-CoV-2 infection impairs mitochondrial function and activates TLR9 signaling in endothelial cells. TLR9 triggers inflammatory responses that lead to endothelial cell dysfunction, potentially contributing to the severity of symptoms in COVID-19. Targeting mitochondrial metabolic pathways may help to define novel therapeutic strategies for COVID-19., (Copyright © 2021. Published by Elsevier Inc.)- Published
- 2022
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29. sTREM-1 Predicts Disease Severity and Mortality in COVID-19 Patients: Involvement of Peripheral Blood Leukocytes and MMP-8 Activity.
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da Silva-Neto PV, de Carvalho JCS, Pimentel VE, Pérez MM, Toro DM, Fraga-Silva TFC, Fuzo CA, Oliveira CNS, Rodrigues LC, Argolo JGM, Carmona-Garcia I, Neto NT, Souza COS, Fernandes TM, Bastos VAF, Degiovani AM, Constant LF, Ostini FM, Feitosa MR, Parra RS, Vilar FC, Gaspar GG, da Rocha JJR, Feres O, Frantz FG, Gerlach RF, Maruyama SR, Russo EMS, Viana AL, Fernandes APM, Santos IKFM, Bonato VLD, Boechat AL, Malheiro A, Sadikot RT, Dias-Baruffi M, Cardoso CRB, Faccioli LH, Sorgi CA, and On Behalf Of The Imunocovid Study Group
- Subjects
- Adolescent, Adult, Aged, Aged, 80 and over, Brazil, Female, Humans, Inflammation, Interleukin-10 blood, Interleukin-6 blood, Interleukin-8 blood, Leukocyte Count, Male, Middle Aged, Neutrophils metabolism, Prospective Studies, SARS-CoV-2, Triggering Receptor Expressed on Myeloid Cells-1 metabolism, Young Adult, Biomarkers blood, COVID-19 diagnosis, COVID-19 mortality, Leukocytes metabolism, Matrix Metalloproteinase 8 metabolism, Severity of Illness Index, Triggering Receptor Expressed on Myeloid Cells-1 blood
- Abstract
Uncontrolled inflammatory responses play a critical role in coronavirus disease (COVID-19). In this context, because the triggering-receptor expressed on myeloid cells-1 (TREM-1) is considered an intrinsic amplifier of inflammatory signals, this study investigated the role of soluble TREM-1 (sTREM-1) as a biomarker of the severity and mortality of COVID-19. Based on their clinical scores, we enrolled COVID-19 positive patients ( n = 237) classified into mild, moderate, severe, and critical groups. Clinical data and patient characteristics were obtained from medical records, and their plasma inflammatory mediator profiles were evaluated with immunoassays. Plasma levels of sTREM-1 were significantly higher among patients with severe disease compared to all other groups. Additionally, levels of sTREM-1 showed a significant positive correlation with other inflammatory parameters, such as IL-6, IL-10, IL-8, and neutrophil counts, and a significant negative correlation was observed with lymphocyte counts. Most interestingly, sTREM-1 was found to be a strong predictive biomarker of the severity of COVID-19 and was related to the worst outcome and death. Systemic levels of sTREM-1 were significantly correlated with the expression of matrix metalloproteinases (MMP)-8, which can release TREM-1 from the surface of peripheral blood cells. Our findings indicated that quantification of sTREM-1 could be used as a predictive tool for disease outcome, thus improving the timing of clinical and pharmacological interventions in patients with COVID-19.
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- 2021
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30. Protective Immunity against Gamma and Zeta Variants after Inactivated SARS-CoV-2 Virus Immunization.
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Fumagalli MJ, Castro-Jorge LA, Fraga-Silva TFC, de Azevedo PO, Capato CF, Rattis BAC, Hojo-Souza NS, Floriano VG, de Castro JT, Ramos SG, da Fonseca BAL, Bonato VLD, Gazzinelli RT, and Figueiredo LTM
- Subjects
- Animals, Antibodies, Neutralizing immunology, Antibodies, Viral immunology, COVID-19 immunology, COVID-19 pathology, COVID-19 virology, COVID-19 Vaccines administration & dosage, Cross Protection, Cytokines metabolism, Follow-Up Studies, Humans, Immunization, Lung metabolism, Lung pathology, Mice, Vaccines, Inactivated administration & dosage, Viral Load, COVID-19 prevention & control, COVID-19 Vaccines immunology, SARS-CoV-2 immunology, Vaccines, Inactivated immunology
- Abstract
The persistent circulation of SARS-CoV-2 represents an ongoing global threat due to the emergence of new viral variants that can sometimes evade the immune system of previously exposed or vaccinated individuals. We conducted a follow-up study of adult individuals that had received an inactivated SARS-CoV-2 vaccine, evaluating antibody production and neutralizing activity over a period of 6 months. In addition, we performed mice immunization with inactivated SARS-CoV-2, and evaluated the immune response and pathological outcomes against Gamma and Zeta variant infection. Vaccinated individuals produced high levels of antibodies with robust neutralizing activity, which was significantly reduced against Gamma and Zeta variants. Production of IgG anti-S antibodies and neutralizing activity robustly reduced after 6 months of vaccination. Immunized mice demonstrated cellular response against Gamma and Zeta variants, and after viral infection, reduced viral loads, IL-6 expression, and histopathological outcome in the lungs. TNF levels were unchanged in immunized or not immunized mice after infection with the Gamma variant. Furthermore, serum neutralization activity rapidly increases after infection with the Gamma and Zeta variants. Our data suggest that immunization with inactivated WT SARS-CoV-2 induces a promptly responsive cross-reactive immunity response against the Gamma and Zeta variants, reducing COVID-19 pathological outcomes.
- Published
- 2021
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31. Artepillin C Reduces Allergic Airway Inflammation by Induction of Monocytic Myeloid-Derived Suppressor Cells.
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Martins NS, de Campos Fraga-Silva TF, Correa GF, Boko MMM, Ramalho LNZ, Rodrigues DM, Hori JI, Costa DL, Bastos JK, and Bonato VLD
- Abstract
Propolis is a natural product produced by bees that is primarily used in complementary and alternative medicine and has anti-inflammatory, antibacterial, antiviral, and antitumoral biological properties. Some studies have reported the beneficial effects of propolis in models of allergic asthma. In a previous study, our group showed that green propolis treatment reduced airway inflammation and mucus secretion in an ovalbumin (OVA)-induced asthma model and resulted in increased regulatory T cells (Treg) and polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC) frequencies in the lungs, two leukocyte populations that have immunosuppressive functions. In this study, we evaluated the anti-inflammatory effects of artepillin C (ArtC), the major compound of green propolis, in the context of allergic airway inflammation. Our results show that ArtC induces in vitro differentiation of Treg cells and monocytic MDSC (M-MDSC). Furthermore, in an OVA-induced asthma model, ArtC treatment reduced pulmonary inflammation, eosinophil influx to the airways, mucus and IL-5 secretion along with increased frequency of M-MDSC, but not Treg cells, in the lungs. Using an adoptive transfer model, we confirmed that the effect of ArtC in the reduction in airway inflammation was dependent on M-MDSC. Altogether, our data show that ArtC exhibits an anti-inflammatory effect and might be an adjuvant therapy for allergic asthma.
- Published
- 2021
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32. Kallikrein 5 Inhibition by the Lympho-Epithelial Kazal-Type Related Inhibitor Hinders Matriptase-Dependent Carcinogenesis.
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da Silva EZM, Fraga-Silva TFC, Yuan Y, Alves MG, Publio GA, da Fonseca CK, Kodama MH, Vieira GV, Candido MF, Innocentini LMAR, Miranda MG, da Silva AR, Alves-Filho JC, Bonato VLD, Iglesias-Bartolome R, and Sales KU
- Abstract
Head and neck squamous cell carcinoma remains challenging to treat with no improvement in survival rates over the past 50 years. Thus, there is an urgent need to discover more reliable therapeutic targets and biomarkers for HNSCC. Matriptase, a type-II transmembrane serine protease, induces malignant transformation in epithelial stem cells through proteolytic activation of pro-HGF and PAR-2, triggering PI3K-AKT-mTOR and NFKB signaling. The serine protease inhibitor lympho-epithelial Kazal-type-related inhibitor (LEKTI) inhibits the matriptase-driven proteolytic pathway, directly blocking kallikreins in epithelial differentiation. Hence, we hypothesized LEKTI could inhibit matriptase-dependent squamous cell carcinogenesis, thus implicating kallikreins in this process. Double-transgenic mice with simultaneous expression of matriptase and LEKTI under the keratin-5 promoter showed a prominent rescue of K5-Matriptase
+/0 premalignant phenotype. Notably, in DMBA-induced SCC, heterotopic co-expression of LEKTI and matriptase delayed matriptase-driven tumor incidence and progression. Co-expression of LEKTI reverted altered Kallikrein-5 expression observed in the skin of K5-Matriptase+/0 mice, indicating that matriptase-dependent proteolytic pathway inhibition by LEKTI occurs through kallikreins. Moreover, we showed that Kallikrein-5 is necessary for PAR-2-mediated IL-8 release, YAP1-TAZ/TEAD activation, and matriptase-mediated oral squamous cell carcinoma migration. Collectively, our data identify a third signaling pathway for matriptase-dependent carcinogenesis in vivo. These findings are critical for the identification of more reliable biomarkers and effective therapeutic targets in Head and Neck cancer.- Published
- 2021
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33. Obesity-Induced Dysbiosis Exacerbates IFN-γ Production and Pulmonary Inflammation in the Mycobacterium tuberculosis Infection.
- Author
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Palma Albornoz SP, Fraga-Silva TFC, Gembre AF, de Oliveira RS, de Souza FM, Rodrigues TS, Kettelhut IDC, Manca CS, Jordao AA, Ramalho LNZ, Ribolla PEM, Carlos D, and Bonato VLD
- Subjects
- Adaptive Immunity, Animals, Bacterial Load, Disease Susceptibility, Dysbiosis immunology, Fecal Microbiota Transplantation, Feces microbiology, Female, Leukocytes metabolism, Lung immunology, Lung microbiology, Lung pathology, Mice, Inbred C57BL, Microbiota, Obesity immunology, Pneumonia immunology, Tuberculosis immunology, Mice, Dysbiosis etiology, Dysbiosis microbiology, Interferon-gamma biosynthesis, Obesity complications, Obesity microbiology, Pneumonia microbiology, Tuberculosis complications
- Abstract
The microbiota of the gut-lung axis affects local and far-reaching immune responses and might also trigger chronic and inflammatory diseases. We hypothesized that gut dysbiosis induced by obesity, which coexists in countries with a high tuberculosis burden, aggravates the host susceptibility and the pulmonary damage tolerance. To assess our hypothesis, we used a model of high-fat diet (HFD)-induced obesity, followed by infection of C57BL/6 mice with Mycobacterium tuberculosis . We showed that obesity increased the susceptibility, the pulmonary inflammation and IFN-γ levels in M. tuberculosis -infected mice. During the comorbidity obesity and tuberculosis, there is an increase of Bacteroidetes and Firmicutes in the lungs, and an increase of Firmicutes and butyrate in the feces. Depletion of gut microbiota by antibiotic treatment in the obese infected mice reduced the frequencies of CD4
+ IFN-γ+ IL-17- cells and IFN-γ levels in the lungs, associated with an increase of Lactobacillus . Our findings reinforce the role of the gut-lung axis in chronic infections and suggest that the gut microbiota modulation may be a potential host-directed therapy as an adjuvant to treat TB in the context of IFN-γ-mediated immunopathology.- Published
- 2021
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34. Heparin prevents in vitro glycocalyx shedding induced by plasma from COVID-19 patients.
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Potje SR, Costa TJ, Fraga-Silva TFC, Martins RB, Benatti MN, Almado CEL, de Sá KSG, Bonato VLD, Arruda E, Louzada-Junior P, Oliveira RDR, Zamboni DS, Becari C, Auxiliadora-Martins M, and Tostes RC
- Subjects
- Aged, Blood Coagulation Disorders blood, Blood Coagulation Disorders virology, COVID-19 metabolism, COVID-19 Testing, Case-Control Studies, Cell Adhesion physiology, Endothelium, Vascular metabolism, Female, Glycocalyx metabolism, Glycocalyx virology, Human Umbilical Vein Endothelial Cells, Humans, Interleukin-1beta blood, Interleukin-6 blood, Male, Middle Aged, Oxidation-Reduction, SARS-CoV-2, Thrombosis metabolism, COVID-19 blood, COVID-19 pathology, Glycocalyx pathology, Heparin pharmacology
- Abstract
The severe forms and worsened outcomes of COVID-19 (coronavirus disease 19) are closely associated with hypertension and cardiovascular disease. Endothelial cells express Angiotensin-Converting Enzyme 2 (ACE2), which is the entrance door for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The hallmarks of severe illness caused by SARS-CoV-2 infection are increased levels of IL-6, C-reactive protein, D-dimer, ferritin, neutrophilia and lymphopenia, pulmonary intravascular coagulopathy and microthrombi of alveolar capillaries. The endothelial glycocalyx, a proteoglycan- and glycoprotein-rich layer covering the luminal side of endothelial cells, contributes to vascular homeostasis. It regulates vascular tonus and permeability, prevents thrombosis, and modulates leukocyte adhesion and inflammatory response. We hypothesized that cytokine production and reactive oxygen species (ROS) generation associated with COVID-19 leads to glycocalyx degradation. A cohort of 20 hospitalized patients with a confirmed COVID-19 diagnosis and healthy subjects were enrolled in this study. Mechanisms associated with glycocalyx degradation in COVID-19 were investigated. Increased plasma concentrations of IL-6 and IL1-β, as well as increased lipid peroxidation and glycocalyx components were detected in plasma from COVID-19 patients compared to plasma from healthy subjects. Plasma from COVID-19 patients induced glycocalyx shedding in cultured human umbilical vein endothelial cells (HUVECs) and disrupted redox balance. Treatment of HUVECs with low molecular weight heparin inhibited the glycocalyx perturbation. In conclusion, plasma from COVID-19 patients promotes glycocalyx shedding and redox imbalance in endothelial cells, and heparin treatment potentially inhibits glycocalyx disruption., (Copyright © 2021 Elsevier Inc. All rights reserved.)
- Published
- 2021
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35. COVID-19: Integrating the Complexity of Systemic and Pulmonary Immunopathology to Identify Biomarkers for Different Outcomes.
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Fraga-Silva TFC, Maruyama SR, Sorgi CA, Russo EMS, Fernandes APM, de Barros Cardoso CR, Faccioli LH, Dias-Baruffi M, and Bonato VLD
- Subjects
- Humans, Lung virology, SARS-CoV-2, Biomarkers analysis, COVID-19 immunology, COVID-19 pathology, Lung immunology, Lung pathology
- Abstract
In the last few months, the coronavirus disease 2019 (COVID-19) pandemic has affected millions of people worldwide and has provoked an exceptional effort from the scientific community to understand the disease. Clinical evidence suggests that severe COVID-19 is associated with both dysregulation of damage tolerance caused by pulmonary immunopathology and high viral load. In this review article, we describe and discuss clinical studies that show advances in the understanding of mild and severe illness and we highlight major points that are critical for improving the comprehension of different clinical outcomes. The understanding of pulmonary immunopathology will contribute to the identification of biomarkers in an attempt to classify mild, moderate, severe and critical COVID-19 illness. The interface of pulmonary immunopathology and the identification of biomarkers are critical for the development of new therapeutic strategies aimed to reduce the systemic and pulmonary hyperinflammation in severe COVID-19., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Fraga-Silva, Maruyama, Sorgi, Russo, Fernandes, de Barros Cardoso, Faccioli, Dias-Baruffi and Bonato.)
- Published
- 2021
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36. Interactions of Extracellular Vesicles from Pathogenic Fungi with Innate Leukocytes.
- Author
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Freitas MS, Pessoni AM, Coelho C, Bonato VLD, Rodrigues ML, Casadevall A, and Almeida F
- Subjects
- Fungi, Humans, Leukocytes, Extracellular Vesicles, Mycoses
- Abstract
Several studies have shown the immunomodulatory effects of extracellular vesicles (EVs) released by pathogenic fungi. Herein, we discuss the data regarding the immunomodulatory properties of fungal EVs, but also of EVs produced by infected leukocytes. This characterizes a two-way path, in which both host and fungal EVs could coexist and play crucial roles in disease progression or protection in fungal infections. We suggest that EVs can dictate the progress of fungal diseases, and their potential as therapeutic targets., (© 2021. Springer Nature Switzerland AG.)
- Published
- 2021
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37. Mycobacterium tuberculosis-infected alveolar epithelial cells modulate dendritic cell function through the HIF-1α-NOS2 axis.
- Author
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Rodrigues TS, Alvarez ARP, Gembre AF, Forni MFPAD, de Melo BMS, Alves Filho JCF, Câmara NOS, and Bonato VLD
- Subjects
- Alveolar Epithelial Cells microbiology, Alveolar Epithelial Cells pathology, Animals, Dendritic Cells microbiology, Dendritic Cells pathology, Hypoxia-Inducible Factor 1, alpha Subunit genetics, Mice, Mice, Knockout, Nitric Oxide Synthase Type II genetics, Tuberculosis, Pulmonary genetics, Tuberculosis, Pulmonary microbiology, Alveolar Epithelial Cells immunology, Dendritic Cells immunology, Hypoxia-Inducible Factor 1, alpha Subunit immunology, Mycobacterium tuberculosis immunology, Nitric Oxide Synthase Type II immunology, Tuberculosis, Pulmonary immunology
- Abstract
Tuberculosis kills more than 1 million people every year, and its control depends on the effective mechanisms of innate immunity, with or without induction of adaptive immune response. We investigated the interaction of type II alveolar epithelial cells (AEC-II) infected by Mycobacterium tuberculosis with dendritic cells (DCs). We hypothesized that the microenvironment generated by this interaction is critical for the early innate response against mycobacteria. We found that AEC-II infected by M. tuberculosis induced DC maturation, which was negatively regulated by HIF-1α-inducible NOS2 axis, and switched DC metabolism from an early and short peak of glycolysis to a low energetic status. However, the infection of DCs by M. tuberculosis up-regulated NOS2 expression and inhibited AEC-II-induced DC maturation. Our study demonstrated, for the first time, that HIF-1α-NOS2 axis plays a negative role in the maturation of DCs during M. tuberculosis infection. Such modulation might be useful for the exploitation of molecular targets to develop new therapeutic strategies against tuberculosis., (©2020 Society for Leukocyte Biology.)
- Published
- 2020
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38. Interplay between alveolar epithelial and dendritic cells and Mycobacterium tuberculosis.
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Rodrigues TS, Conti BJ, Fraga-Silva TFC, Almeida F, and Bonato VLD
- Subjects
- Alveolar Epithelial Cells pathology, Dendritic Cells pathology, Humans, Macrophages, Alveolar immunology, Macrophages, Alveolar pathology, Tuberculosis, Pulmonary pathology, Adaptive Immunity, Alveolar Epithelial Cells immunology, Dendritic Cells immunology, Host-Pathogen Interactions immunology, Mycobacterium tuberculosis physiology, Tuberculosis, Pulmonary immunology
- Abstract
The innate response plays a crucial role in the protection against tuberculosis development. Moreover, the initial steps that drive the host-pathogen interaction following Mycobacterium tuberculosis infection are critical for the development of adaptive immune response. As alveolar Mϕs, airway epithelial cells, and dendritic cells can sense the presence of M. tuberculosis and are the first infected cells. These cells secrete mediators, which generate inflammatory signals that drive the differentiation and activation of the T lymphocytes necessary to clear the infection. Throughout this review article, we addressed the interaction between epithelial cells and M. tuberculosis, as well as the interaction between dendritic cells and M. tuberculosis. The understanding of the mechanisms that modulate those interactions is critical to have a complete view of the onset of an infection and may be useful for the development of dendritic cell-based vaccine or immunotherapies., (©2020 Society for Leukocyte Biology.)
- Published
- 2020
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39. NOD2 Deficiency Promotes Intestinal CD4+ T Lymphocyte Imbalance, Metainflammation, and Aggravates Type 2 Diabetes in Murine Model.
- Author
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Carlos D, Pérez MM, Leite JA, Rocha FA, Martins LMS, Pereira CA, Fraga-Silva TFC, Pucci TA, Ramos SG, Câmara NOS, Bonato VLD, Tostes RC, and Silva JS
- Subjects
- Animals, Biomarkers, Diet, High-Fat, Disease Models, Animal, Gastrointestinal Microbiome immunology, Gene Expression Profiling, Glucose metabolism, Immunohistochemistry, Insulin blood, Insulin metabolism, Intestinal Mucosa immunology, Intestinal Mucosa metabolism, Islets of Langerhans metabolism, Leukocytes immunology, Leukocytes metabolism, Lipid Metabolism, Mice, Mice, Knockout, Obesity etiology, Obesity metabolism, Permeability, Signal Transduction, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes metabolism, Diabetes Mellitus, Type 2 etiology, Diabetes Mellitus, Type 2 metabolism, Inflammation etiology, Inflammation metabolism, Nod2 Signaling Adaptor Protein deficiency
- Abstract
Type 2 diabetes (T2D) is a metabolic disease characterized by increased inflammation, NOD-like receptors (NLRs) activation and gut dysbiosis. Our research group has recently reported that intestinal Th17 response limits gut dysbiosis and LPS translocation to visceral adipose tissue (VAT), protecting against metabolic syndrome. However, whether NOD2 receptor contributes intestinal Th17 immunity, modulates dysbiosis-driven metabolic tissue inflammation, and obesity-induced T2D remain poorly understood. In this context, we observed that mice lacking NOD2 fed a high-fat diet (HFD) display severe obesity, exhibit greater adiposity, and more hepatic steatosis compared to HFD-fed wild-type (WT) mice. In addition, they develop increased hyperglycemia, worsening of glucose intolerance, and insulin resistance. Notably, the deficiency of NOD2 causes a deviation from M2 macrophage and regulatory T cells (Treg) to M1 macrophage and mast cells into VAT compared to WT mice fed HFD. An imbalance was also observed in Th17/Th1 cell populations, with reduced IL-17 and IL-22 gene expression in the mesenteric lymph nodes (MLNs) and ileum, respectively, of NOD2-deficient mice fed HFD. 16S rRNA sequencing indicates lower richness, alpha diversity, and a depletion of Allobaculum, Lactobacillus , and enrichment with Bacteroides genera in these mice compared to HFD-fed WT mice. These alterations were associated with disrupted tight-junctions expression, augmented serum LPS, and bacterial translocation into VAT. Overall, NOD2 activation is required for a protective Th17 over Th1 immunity in the gut, which seems to decrease gram-negative bacteria outgrowth in gut microbiota, attenuating the endotoxemia, metainflammation, and protecting against obesity-induced T2D., (Copyright © 2020 Carlos, Pérez, Leite, Rocha, Martins, Pereira, Fraga-Silva, Pucci, Ramos, Câmara, Bonato, Tostes and Silva.)
- Published
- 2020
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40. Pathogenic Allodiploid Hybrids of Aspergillus Fungi.
- Author
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Steenwyk JL, Lind AL, Ries LNA, Dos Reis TF, Silva LP, Almeida F, Bastos RW, Fraga da Silva TFC, Bonato VLD, Pessoni AM, Rodrigues F, Raja HA, Knowles SL, Oberlies NH, Lagrou K, Goldman GH, and Rokas A
- Subjects
- Aspergillus isolation & purification, Diploidy, Genomics, Aspergillus genetics, Genome, Fungal, Hybridization, Genetic
- Abstract
Interspecific hybridization substantially alters genotypes and phenotypes and can give rise to new lineages. Hybrid isolates that differ from their parental species in infection-relevant traits have been observed in several human-pathogenic yeasts and plant-pathogenic filamentous fungi but have yet to be found in human-pathogenic filamentous fungi. We discovered 6 clinical isolates from patients with aspergillosis originally identified as Aspergillus nidulans (section Nidulantes) that are actually allodiploid hybrids formed by the fusion of Aspergillus spinulosporus with an unknown close relative of Aspergillus quadrilineatus, both in section Nidulantes. Evolutionary genomic analyses revealed that these isolates belong to Aspergillus latus, an allodiploid hybrid species. Characterization of diverse infection-relevant traits further showed that A. latus hybrid isolates are genomically and phenotypically heterogeneous but also differ from A. nidulans, A. spinulosporus, and A. quadrilineatus. These results suggest that allodiploid hybridization contributes to the genomic and phenotypic diversity of filamentous fungal pathogens of humans., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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41. Green propolis increases myeloid suppressor cells and CD4 + Foxp3 + cells and reduces Th2 inflammation in the lungs after allergen exposure.
- Author
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Piñeros AR, de Lima MHF, Rodrigues T, Gembre AF, Bertolini TB, Fonseca MD, Berretta AA, Ramalho LNZ, Cunha FQ, Hori JI, and Bonato VLD
- Subjects
- Allergens, Animals, Anti-Inflammatory Agents pharmacology, Asthma chemically induced, Asthma immunology, Bronchoalveolar Lavage Fluid cytology, Bronchoalveolar Lavage Fluid immunology, Cell Differentiation drug effects, Female, Immunologic Factors pharmacology, Immunotherapy, Interleukin-13 genetics, Interleukin-13 immunology, Interleukin-5 immunology, Lung drug effects, Lung immunology, Mice, Inbred C57BL, Myeloid-Derived Suppressor Cells immunology, Ovalbumin, Propolis pharmacology, T-Lymphocytes, Regulatory immunology, Th2 Cells drug effects, Th2 Cells immunology, Anti-Inflammatory Agents therapeutic use, Asthma drug therapy, Immunologic Factors therapeutic use, Myeloid-Derived Suppressor Cells drug effects, Propolis therapeutic use, T-Lymphocytes, Regulatory drug effects
- Abstract
Ethnopharmacological Relevance: Propolis is a natural product produced by honeybees used as a medicine at least to 300 BC. In the last decades, several studies showed biological and pharmacological properties of propolis, witch scientifically explains the empirical use for centuries. The anti-inflammatory activity of propolis with the purpose to reduce Th2 inflammation has been evaluated in allergic asthma. However, it remains to be determined how propolis negatively regulates the immune response after allergen re-exposure., Aim of the Study: We hypothesized that the anti-inflammatory activity of propolis is dependent on the induction of myeloid derived suppressor cells (MDSC) and regulatory T cells., Materials and Methods: To assess this hypothesis, we used an ovalbumin-induced asthma model to evaluate the effect of EPP-AF® dry extract from Brazilian green propolis., Results: Propolis treatment decreased pulmonary inflammation and mucus production as well as eosinophils and IL-5 in the broncoalveolar lavage. Propolis enhanced also in vitro differentiation and in vivo frequency of lung MDSC and CD4
+ Foxp3+ regulatory T cells., Conclusions: Together these results confirm the immunomodulatory potential of propolis during sensitization and challenge with allergen. In addition, the collecting findings show, for the first time, that propolis increases the frequency of MDSC and CD4+ Foxp3+ regulatory T cells in the lungs, and suggest that it could be use as target for development of new immunotherapy or adjuvant immunotherapy for asthma., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to disclose., (Copyright © 2019. Published by Elsevier B.V.)- Published
- 2020
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42. Fungal Extracellular Vesicles as Potential Targets for Immune Interventions.
- Author
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Freitas MS, Bonato VLD, Pessoni AM, Rodrigues ML, Casadevall A, and Almeida F
- Subjects
- Mycoses immunology, Mycoses microbiology, Extracellular Vesicles metabolism, Fungi metabolism, Host-Pathogen Interactions, Immunologic Factors metabolism
- Abstract
The release of extracellular vesicles (EVs) by fungi is a fundamental cellular process. EVs carry several biomolecules, including pigments, proteins, enzymes, lipids, nucleic acids, and carbohydrates, and are involved in physiological and pathological processes. EVs may play a pivotal role in the establishment of fungal infections, as they can interact with the host immune system to elicit multiple outcomes. It has been observed that, depending on the fungal pathogen, EVs can exacerbate or attenuate fungal infections. The study of the interaction between fungal EVs and the host immune system and understanding of the mechanisms that regulate those interactions might be useful for the development of new adjuvants as well as the improvement of protective immune responses against infectious or noninfectious diseases. In this review, we describe the immunomodulatory properties of EVs produced by pathogenic fungi and discuss their potential as adjuvants for prophylactic or therapeutic strategies., (Copyright © 2019 Freitas et al.)
- Published
- 2019
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43. CCR4-dependent reduction in the number and suppressor function of CD4 + Foxp3 + cells augments IFN-γ-mediated pulmonary inflammation and aggravates tuberculosis pathogenesis.
- Author
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Bertolini TB, Piñeros AR, Prado RQ, Gembre AF, Ramalho LNZ, Alves-Filho JC, and Bonato VLD
- Subjects
- Animals, CD4-Positive T-Lymphocytes pathology, Female, Forkhead Transcription Factors genetics, Forkhead Transcription Factors immunology, Interferon-gamma genetics, Mice, Mice, Knockout, Receptors, CCR4 genetics, Tuberculosis, Pulmonary genetics, Tuberculosis, Pulmonary pathology, CD4-Positive T-Lymphocytes immunology, Interferon-gamma immunology, Mycobacterium tuberculosis immunology, Receptors, CCR4 immunology, Tuberculosis, Pulmonary immunology
- Abstract
Chronic pulmonary inflammation marked predominantly by CD4
+ IFN-γ+ cells is the hallmark of tuberculosis pathogenesis in immunocompetent adults, who are substantially affected by this disease. Moreover, CD4+ Foxp3+ cell-mediated suppression contributes to infection susceptibility. We addressed the role of CD4+ Foxp3+ cells in tuberculosis pathogenesis, because this aspect has not been addressed during chronic infection. We targeted CCR4, which induces the influx of CD4+ Foxp3+ cells into the lungs. CCR4-/- mice exhibited a lower frequency of CD4+ Foxp3+ cells at 15, 30, and 70 days of infection than their wild-type counterparts. However, only at 70 days of infection was an exacerbated IFN-γ-mediated immune response associated with apparent tuberculosis pathogenesis and susceptibility. In addition, CCR4-/- mice exhibited a decrease in the suppressor function of CD4+ Foxp3+ cells. Adoptive transfer of Foxp3+ cells into infected CCR4-/- mice restored pulmonary inflammation and bacterial load to levels observed in wild-type mice. Our findings suggest that CD4+ Foxp3+ cells play a time-dependent role in tuberculosis and highlight that CCR4 plays a critical role in the balance of IFN-γ-mediated inflammation by regulating the influx and function of CD4+ Foxp3+ cells. Our findings are translationally relevant, as CD4+ Foxp3+ cells or CCR4 could be a target for immunotherapy, considering the heterogeneity of tuberculosis in immunocompetent adults.- Published
- 2018
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44. Systemic Immunological changes in patients with distinct clinical outcomes during Mycobacterium tuberculosis infection.
- Author
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Morais-Papini TF, Coelho-Dos-Reis JGA, Wendling APB, do Vale Antonelli LR, Wowk PF, Bonato VLD, Augusto VM, Elói-Santos S, Martins-Filho OA, Carneiro CM, and Teixeira-Carvalho A
- Subjects
- Adult, Aged, Antigens, CD19 metabolism, CD56 Antigen metabolism, Cytokines metabolism, Female, HLA-DR Antigens metabolism, Humans, Immunity, Leukocytes, Mononuclear microbiology, Lung anatomy & histology, Lung microbiology, Male, Middle Aged, Receptors, Chemokine metabolism, Young Adult, B-Lymphocytes immunology, CD4-Positive T-Lymphocytes immunology, Killer Cells, Natural immunology, Leukocytes, Mononuclear immunology, Lung immunology, Mycobacterium tuberculosis immunology, Tuberculosis immunology
- Abstract
Background: The lung lesions in an individual infected with tuberculosis (TB) are surprisingly variable and independent of each other. However, there is no circulating biomarker yet able to segregate patients according to the extent of lung lesions., Materials and Methods: In this study, the phenotypic and functional profile of leukocytes of patients with active pulmonary tuberculosis (TB) and controls (CO) were fully scrutinized by immunophenotyping assays and in vitro short-term whole blood culture. The TB group was subdivided according to the extent of lung lesions as unilateral (UNI) and bilateral (BI)., Results: The results show that TB group display an altered leukocyte profile in the peripheral blood with significant lower counts of NK-cells, CD3
+ CD56+ CD16+/- NKT-cells, CD4+ T-cells, CD19+ B-cells when compared to CO. Increased CD4+ T-cells and CD8+ T-cell activation was observed by the upregulation of activation markers (HLA-DR) as well as of chemokine receptors (CCR2, CCR3, and CXCR4). In addition, TB group presented a significant decrease proportion of CD14Low CD16+ monocytes despite the increase in HLA-DR expression. Regarding the severity of the disease, in the BI group a reduction in frequency of CD19+ CD5+ B-cells and expression of HLA-DR in CD14Low CD16+ monocytes was observed. Furthermore, the extent of lung lesions influences the production of molecules as observed by significantly larger production of IL-4 by neutrophils, total T-cells, CD4+ T-cells, CD8+ T-cells and CD19+ B-cells in UNI as compared to BI. By contrast, in BI group the frequency of high producers of both IL-17+ CD4+ T-cells and IL-17+ CD8+ T-cells were significantly increased than UNI, suggesting the deleterious role of these subsets during active pulmonary Mtb infection., Conclusion: The immunophenotypic characterization of unilateral and bilateral active TB performed in the present study indicates that the extent of lung lesion could be associated with a fine-tuning between immunological responses during untreated Mtb infection., (Copyright © 2017 Elsevier GmbH. All rights reserved.)- Published
- 2017
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45. Oral administration of Saccharomyces cerevisiae UFMG A-905 prevents allergic asthma in mice.
- Author
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Fonseca VMB, Milani TMS, Prado R, Bonato VLD, Ramos SG, Martins FS, Vianna EO, and Borges MC
- Subjects
- Administration, Oral, Animals, Asthma prevention & control, Bronchoalveolar Lavage Fluid, Disease Models, Animal, Eosinophils immunology, Interferon-gamma immunology, Interleukin-10 immunology, Interleukin-13 immunology, Interleukin-17 immunology, Interleukin-4 immunology, Lung immunology, Lung physiopathology, Male, Mice, Mice, Inbred BALB C, Ovalbumin, Respiratory Hypersensitivity prevention & control, Asthma immunology, Probiotics, Respiratory Hypersensitivity immunology, Saccharomyces cerevisiae
- Abstract
Background and Objective: The prevalence of asthma has increased in communities that adopt a Western lifestyle and become more urbanized. Probiotics may be effective in the prevention of allergic diseases, such as asthma. The aim of the current study was to examine the effects of Saccharomyces cerevisiae UFMG A-905 in an allergic model of asthma., Methods: Balb/c mice were sensitized twice with ovalbumin (OVA) intraperitoneally, 1 week apart and challenged with OVA intranasally for 3 days. Mice were daily treated with S. cerevisiae UFMG A-905 via gavaging needle 10 days before OVA sensitization and during challenges. After challenge, in vivo lung function was measured, and bronchoalveolar lavage (BAL) and lung inflammation were assessed., Results: Oral treatment with S. cerevisiae UFMG A-905 significantly decreased airway hyperresponsiveness, total cell number and the influx of eosinophils to the airway, inflammatory cell in the lung, mucus expression in epithelial cells and the levels of IL-4, IL-5 and IL-13. Additionally, S. cerevisiae UFMG A-905 restored the levels of IL-10 and interferon (IFN)-gamma, and increased the levels of IL-17A., Conclusion: Oral administration of S. cerevisiae UFMG A-905 prevented the development of major asthma-like characteristics in a mouse model., (© 2017 Asian Pacific Society of Respirology.)
- Published
- 2017
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46. Mycobacterial Hsp65 antigen upregulates the cellular immune response of healthy individuals compared with tuberculosis patients.
- Author
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Wowk PF, Franco LH, Fonseca DMD, Paula MO, Vianna ÉDSO, Wendling AP, Augusto VM, Elói-Santos SM, Teixeira-Carvalho A, Silva FDC, Vinhas SA, Martins-Filho OA, Palaci M, Silva CL, and Bonato VLD
- Subjects
- Adult, Bacterial Proteins genetics, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Chaperonin 60 genetics, Cytotoxicity, Immunologic, Female, Healthy Volunteers, Humans, Interferon-gamma biosynthesis, Interferon-gamma immunology, Interleukin-10 biosynthesis, Interleukin-10 immunology, Leukocytes, Mononuclear immunology, Macrophages, Alveolar immunology, Male, Middle Aged, Mycobacterium tuberculosis immunology, Recombinant Proteins immunology, Tuberculosis Vaccines immunology, Up-Regulation, Vaccines, DNA pharmacology, Young Adult, Bacterial Proteins immunology, Chaperonin 60 immunology, Immunity, Cellular, Lymphocyte Activation, Tuberculosis immunology
- Abstract
Previously we showed that 65-kDa Mycobacterium leprae heat shock protein (Hsp65) is a target for the development of a tuberculosis vaccine. Here we evaluated peripheral blood mononuclear cells (PBMC) from healthy individuals or tuberculosis patients stimulated with two forms of Hsp65 antigen, recombinant DNA that encodes Hsp65 (DNA-HSP65) or recombinant Hsp65 protein (rHsp65) in attempting to mimic a prophylactic or therapeutic study in vitro, respectively. Proliferation and cytokine-producing CD4
+ or CD8+ cell were assessed by flow cytometry. The CD4+ cell proliferation from healthy individuals was stimulated by DNA-HSP65 and rHsp65, while CD8+ cell proliferation from healthy individuals or tuberculosis patients was stimulated by rHSP65. DNA-HSP65 did not improve the frequency of IFN-gamma+ cells from healthy individuals or tuberculosis patients. Furthermore, we found an increase in the frequency of IL-10-producing cells in both groups. These findings show that Hsp65 antigen activates human lymphocytes and plays an immune regulatory role that should be addressed as an additional antigen for the development of antigen-combined therapies.- Published
- 2017
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