Pharmacological blockade of protease-Activated Receptor 2 improves airway remodeling and lung inflammation in experimental allergic asthma

Protease-activated receptors (PARs) are metabotropic G-protein-coupled receptors that are activated via proteolytic cleavage of a specific sequence of amino acids in their N-terminal region. PAR2 has been implicated in mediating allergic airway inflammation. This study aims to study the effect of PAR2 antagonist ENMD1068in lung inflammation and airway remodeling in experimental asthma. Allergic lung inflammation was induced in sensitized BALB/c mice through intranasal instillations of ovalbumin (OVA), and mice were pretreated with ENMD1068 1 hour before each OVA challenge. Bronchoalveolar lavage fluid (BALF) was collected, and the lungs were removed at different time intervals after OVA challenge to analyze inflammation, airway remodeling and airway hyperresponsiveness. Ovalbumin promoted leukocyte infiltration into BALF in a PAR2-dependent manner. ENMD1068 impaired eosinophil peroxidase (EPO) and myeloperoxidase (MPO) activity in the lung parenchyma into BALF and reduced the loss of dynamic pulmonary compliance, lung resistance in response to methacholine, mucus production, collagen deposition and chemokine (C-C motif) ligand 5 expression compared to those in OVA-challenged mice. We propose that proteases released after an allergen challenge may be crucial to the development of allergic asthma in mice, and PAR2 blockade may be useful as a new pharmacological approach for the treatment of airway allergic diseases

The Therapeutic Treatment with the GAG-Binding Chemokine Fragment CXCL9(74–103) Attenuates Neutrophilic Inflammation and Lung Dysfunction during <i>Klebsiella pneumoniae</i> Infection in Mice

Klebsiella pneumoniae is an important pathogen associated with hospital-acquired pneumonia (HAP). Bacterial pneumonia is characterized by a harmful inflammatory response with a massive influx of neutrophils, production of cytokines and chemokines, and consequent tissue damage and dysfunction. Targeted therapies to block neutrophil migration to avoid tissue damage while keeping the antimicrobial properties of tissue remains a challenge in the field. Here we tested the effect of the anti-inflammatory properties of the chemokine fragment CXCL9(74–103) in pneumonia induced by Klebsiella pneumoniae in mice. Mice were infected by intratracheal injection of Klebsiella pneumoniae and 6 h after infection were treated systemically with CXCL9(74–103). The recruitment of leukocytes, levels of cytokines and chemokines, colony-forming units (CFU), and lung function were evaluated. The treatment with CXCL9(74–103) decreased neutrophil migration to the airways and the production of the cytokine interleukin-1β (IL-1β) without affecting bacterial control. In addition, the therapeutic treatment improved lung function in infected mice. Our results indicated that the treatment with CXCL9(74–103) reduced inflammation and improved lung function in Klebsiella pneumoniae-induced pneumonia

Absence of CCR2 Promotes Proliferation of Alveolar Macrophages That Control Lung Inflammation in Acute Respiratory Distress Syndrome in Mice

Acute respiratory distress syndrome (ARDS) consists of uncontrolled inflammation that causes hypoxemia and reduced lung compliance. Since it is a complex process, not all details have been elucidated yet. In a well-controlled experimental murine model of lipopolysaccharide (LPS)-induced ARDS, the activity and viability of macrophages and neutrophils dictate the beginning and end phases of lung inflammation. C-C chemokine receptor type 2 (CCR2) is a critical chemokine receptor that mediates monocyte/macrophage activation and recruitment to the tissues. Here, we used CCR2-deficient mice to explore mechanisms that control lung inflammation in LPS-induced ARDS. CCR2&minus;/&minus; mice presented higher total numbers of pulmonary leukocytes at the peak of inflammation as compared to CCR2+/+ mice, mainly by enhanced influx of neutrophils, whereas we observed two to six-fold lower monocyte or interstitial macrophage numbers in the CCR2&minus;/&minus;. Nevertheless, the time needed to control the inflammation was comparable between CCR2+/+ and CCR2&minus;/&minus;. Interestingly, CCR2&minus;/&minus; mice presented higher numbers and increased proliferative rates of alveolar macrophages from day 3, with a more pronounced M2 profile, associated with transforming growth factor (TGF)-&beta; and C-C chemokine ligand (CCL)22 production, decreased inducible nitric oxide synthase (Nos2), interleukin (IL)-1&beta; and IL-12b mRNA expression and increased mannose receptor type 1 (Mrc1) mRNA and CD206 protein expression. Depletion of alveolar macrophages significantly delayed recovery from the inflammatory insult. Thus, our work shows that the lower number of infiltrating monocytes in CCR2&minus;/&minus; is partially compensated by increased proliferation of resident alveolar macrophages during the inflammation control of experimental ARDS

Ovarian Grafts 10 Days after Xenotransplantation: Folliculogenesis and Recovery of Viable Oocytes.

Ovarian xenotransplantation is a promising alternative to preserve fertility of oncologic patients. However, several functional aspects of this procedure remained to be addressed. The aim of this study was evaluate the feasibility of xenotransplantation as a strategy to maintain bovine ovarian grafts and produce oocytes. Adult ovarian cortical pieces were xenotransplanted to the dorsal subcutaneous of female NOD-SCID mice (n = 62). Grafts were recovered ten days after xenotransplantation. Host and graft weights; folliculogenesis progression; blood perfusion, relative gene expression and number of macrophage and neutrophil of xenografts; in vitro developmental competence of graft-derived oocytes were evaluated. Folliculogenesis was supported in the grafts, as indicated by the presence of primordial, primary, secondary, antral, and atretic follicles. The xenografts showed a greater volumetric density of atretic follicles and higher hyperemia and number of host-derived macrophage and neutrophil (P<0.05), when compared to non-grafted fragments. There was a higher blood perfusion under the back skin in the transplantation sites of host animals than in control and non-grafted (P<0.01). BAX and PRDX1 genes were up-regulated, while BCL2, FSHR, IGF1R and IGF2R were down-regulated, when compared to the control (P<0.01). Twenty seven oocytes were successfully harvested from grafts, and some of these oocytes were able to give rise to blastocysts after in vitro fertilization. However, cleavage and blastocyst rates of xenograft derived oocytes were lower than in control (P<0.01). Despite showing some functional modifications, the ovarian xenografts were able to support folliculogenesis and produce functional oocytes

Versican and Tumor-Associated Macrophages Promotes Tumor Progression and Metastasis in Canine and Murine Models of Breast Carcinoma

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2019-09-30T16:13:26Z No. of bitstreams: 1 Reis, C. D. Versican....pdf: 6212673 bytes, checksum: f0e5f7d54df5fde4549844031d7af599 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2019-09-30T16:50:18Z (GMT) No. of bitstreams: 1 Reis, C. D. Versican....pdf: 6212673 bytes, checksum: f0e5f7d54df5fde4549844031d7af599 (MD5)Made available in DSpace on 2019-09-30T16:50:18Z (GMT). No. of bitstreams: 1 Reis, C. D. Versican....pdf: 6212673 bytes, checksum: f0e5f7d54df5fde4549844031d7af599 (MD5) Previous issue date: 2019Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).Universidade Federal de Minas Gerais. Institute of Biological Sciences. Department of General Pathology. Belo Horizonte, MG, Brazil / Universidade Federal de Minas Gerais. Institute of Biological Sciences. Laboratory of Pulmonary Immunology and Mechanics. Department of Physiology and Biophysics. Belo Horizonte, MG, Brazil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.Universidade Federal de Minas Gerais. Institute of Biological Sciences. Department of General Pathology. Belo Horizonte, MG, Brazil.Universidade Federal de Minas Gerais. Institute of Biological Sciences. Department of General Pathology. Belo Horizonte, MG, Brazil.Universidade Federal de Minas Gerais. Institute of Biological Sciences. Department of General Pathology. Belo Horizonte, MG, Brazil.Universidade Federal de Minas Gerais. Institute of Biological Sciences. Laboratory of Pulmonary Immunology and Mechanics. Department of Physiology and Biophysics. Belo Horizonte, MG, Brazil.Universidade Federal de Minas Gerais. Institute of Biological Sciences. Department of General Pathology. Belo Horizonte, MG, Brazil.Universidade Federal de Minas Gerais. Institute of Biological Sciences. Laboratory of Pulmonary Immunology and Mechanics. Department of Physiology and Biophysics. Belo Horizonte, MG, Brazil.Universidade Federal de Minas Gerais. Institute of Biological Sciences. Department of Biochemistry and Immunology. Belo Horizonte, MG, Brazil.Universidade Federal de Minas Gerais. Institute of Biological Sciences. Department of General Pathology. Belo Horizonte, MG, Brazil.Universidade Federal de Minas Gerais. Institute of Biological Sciences. Department of General Pathology. Belo Horizonte, MG, Brazil.Universidade Federal de Minas Gerais. Institute of Biological Sciences. Laboratory of Pulmonary Immunology and Mechanics. Department of Physiology and Biophysics. Belo Horizonte, MG, Brazil.Universidade Federal de Minas Gerais. Institute of Biological Sciences. Department of General Pathology. Belo Horizonte, MG, Brazil.Versican and tumor-associated macrophages (TAMs) are involved in growth and metastases in several cancers. Here, we investigated the potential role of versican, a matrix proteoglycan, and its correlation with TAMs infiltrates in different stages of two different breast cancer models: spontaneous canine mammary gland carcinomas and the murine 4T1 breast cancer model. The stromal versican expression was correlated with TAMs accumulation in tumors with an advanced stage from spontaneous canine mammary carcinoma samples. Versican expression in mice, identified in late stages of tumor progression, was associated to a high number of peri-tumoral infiltrating TAMs. Indeed, TAMs were related to a pro-inflammatory and pro-angiogenic state in the primary tumor. Furthermore, TAMs accumulation was related to versican expression in the lungs and an increased number of pulmonary metastatic nodules with pulmonary mechanical dysfunction, which was due to leukocyte influx in the airways and elevated growth factor levels in the microenvironment. Thus, we suggest that versican and TAMs as attractive targets for breast cancer therapy

Multiple Exposures to Ascaris suum Induce Tissue Injury and Mixed Th2/Th17 Immune Response in Mice

Submitted by Nuzia Santos ([email protected]) on 2017-01-20T16:39:47Z No. of bitstreams: 1 ve_Nogueira_Denise_Multiple Exposures_CPqRR_2016.pdf: 7283772 bytes, checksum: 9287d516a1c1fa65a4453682a7bd231f (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2017-01-20T17:32:40Z (GMT) No. of bitstreams: 1 ve_Nogueira_Denise_Multiple Exposures_CPqRR_2016.pdf: 7283772 bytes, checksum: 9287d516a1c1fa65a4453682a7bd231f (MD5)Made available in DSpace on 2017-01-20T17:32:40Z (GMT). No. of bitstreams: 1 ve_Nogueira_Denise_Multiple Exposures_CPqRR_2016.pdf: 7283772 bytes, checksum: 9287d516a1c1fa65a4453682a7bd231f (MD5) Previous issue date: 2016Universidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Parasitologia. Laboratorio de Imunologia e Genomica de Parasitos. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Parasitologia. Laboratorio de Imunologia e Genomica de Parasitos. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Parasitologia. Laboratorio de Imunologia e Genomica de Parasitos. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Parasitologia. Laboratorio de Imunologia e Genomica de Parasitos. Belo Horizonte, MG, Brasil/Universidade Federal do Mato Grosso. Instituto de Biologia e Ciencia da Saude. Cuiabá, MT, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Parasitologia. Laboratorio de Imunologia e Genomica de Parasitos. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Parasitologia. Laboratorio de Imunologia e Genomica de Parasitos. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Parasitologia. Laboratorio de Imunologia e Genomica de Parasitos. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Departamento de Patologia Geral. Instituto de Ciencias Biologicas. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Fisiologia e Biofisica Laboratorio de Imunologia e Mecânica Pulmonar. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Fisiologia e Biofisica Laboratorio de Imunologia e Mecânica Pulmonar. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Departamento de Patologia Geral. Instituto de Ciencias Biologicas. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Laboratorio de Imunologia Celular e Molecular. Belo Horizonte, Brasil/University of Nottingham. United KingdomUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Parasitologia. Laboratorio de Imunologia e Genomica de Parasitos. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Fisiologia e Biofisica Laboratorio de Imunologia e Mecânica Pulmonar. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Parasitologia. Laboratorio de Imunologia e Genomica de Parasitos. Belo Horizonte, MG, Brasil/Universidade Federal do Mato Grosso. Instituto de Biologia e Ciencia da Saude. Cuiabá, MT, BrasilAscaris spp. infection affects 800 million people worldwide, and half of the world population is currently at risk of infection. Recurrent reinfection in humans is mostly due to the simplicity of the parasite life cycle, but the impact of multiple exposures to the biology of the infection and the consequences to the host's homeostasis are poorly understood. In this context, single and multiple exposures in mice were performed in order to characterize the parasitological, histopathological, tissue functional and immunological aspects of experimental larval ascariasis. The most important findings revealed that reinfected mice presented a significant reduction of parasite burden in the lung and an increase in the cellularity in the bronchoalveolar lavage (BAL) associated with a robust granulocytic pulmonary inflammation, leading to a severe impairment of respiratory function. Moreover, the multiple exposures to Ascaris elicited an increased number of circulating inflammatory cells as well as production of higher levels of systemic cytokines, mainly IL-4, IL-5, IL-6, IL-10, IL-17A and TNF-α when compared to single-infected animals. Taken together, our results suggest the intense pulmonary inflammation associated with a polarized systemic Th2/Th17 immune response are crucial to control larval migration after multiple exposures to Ascaris

Progression of folliculogenesis, morphometry and inflammatory cell numbers in xenografted and in control ovarian fragments.

<p>(a) Primordial, (b) primary, (c) secondary, (d) antral and (e) atretic follicles were noted 10 days after xenotransplantation. (g) Volumetric density (%) of ovarian pieces before and after xenotransplantation showing a higher incidence of follicular atresia and hyperemia (f; <i>P<</i>0.01) after this procedure. (h, i) As demonstrated using (h) NAG and (i) MPO assays in xenografted ovarian wet tissue, higher concentration of host-derived macrophages and neutrophils were observed (<i>P<</i>0.01). Black arrows = healthy follicles; red arrows = atretic follicles; white arrows = blood vessels; black arrowhead = hyperemia; asterisks = oocytes. Bars in a, b, c and e = 80 μm and d and f = 150 μm.</p

Primer sequences used for relative gene expression analysis by real-time polymerase chain reaction.

<p>Primer sequences used for relative gene expression analysis by real-time polymerase chain reaction.</p

Number of larvae recovered from host organs.

<p>(A) Liver on the 4^th day post-infection; (B) lung on the 8^th day post-infection; (C) BAL on the 8^th day post-infection; and (D) gut on the 12^th day post-infection. Filled circles–single infection (SI) group; Open circles–reinfection (RE) group. Mann-Whitney test was used to assess differences between groups and are depicted in the graphs by the p values.</p

Paradoxical role of matrix metalloproteinases in liver injury and regeneration after sterile acute hepatic failure

Acetaminophen (APAP) poisoning is one of the leading causes of acute hepatic failure and liver transplantation is often the only lifesaving alternative. During the course of hepatocyte necrosis, an intense accumulation of neutrophils is often observed within the liver microenvironment. Despite the classic idea that neutrophil accumulation in tissues causes collateral tissue damage, there is a growing body of evidence showing that neutrophils can also orchestrate the resolution of inflammation. In this work, drug-induced liver injury was induced by oral administration of APAP and pharmacological intervention was made 12 h after this challenge. Liver injury and repair kinetics were evaluated by a novel combination of enzyme quantifications, ELISA, specific antagonists of neutrophil enzymes and confocal intravital microscopy. We have demonstrated that neutrophil infiltration is not only involved in injury amplification, but also in liver tissue repair after APAP-induced liver injury. In fact, while neutrophil depletion led to reduced hepatic necrosis during APAP poisoning, injury recovery was also delayed in neutropenic mice. The mechanisms underlying the neutrophil reparative role involved rapid degranulation and matrix metalloproteinases (MMPs) activity. Our data highlights the crucial role of neutrophils, in particular for MMPs, in the resolution phase of APAP-induced inflammatory response