Potential of PEGylated toll-like receptor 7 ligands for controlling inflammation and functional changes in mouse models of asthma and silicosis

Prior investigations show that signaling activation through pattern recognition receptors can directly impact a number of inflammatory lung diseases. While toll-like receptor (TLR) 7 agonists have raised interest for their ability to inhibit allergen-induced pathological changes in experimental asthma conditions, the putative benefit of this treatment is limited by adverse effects. Our aim was to evaluate the therapeutic potential of two PEGylated purine-like compounds, TMX-302 and TMX-306, characterized by TLR7 partial agonistic activity; therefore, the compounds are expected to induce lower local and systemic adverse reactions. In vitro approaches and translation to murine models of obstructive and restrictive lung diseases were explored. In vitro studies with human PBMCs showed that both TMX-302 and TMX-306 marginally affects cytokine production as compared with equivalent concentrations of the TLR7 full agonist, TMX-202. The PEGylated compounds did not induce monocyte-derived DC maturation or B cell proliferation, differently from what observed after stimulation with TMX-202. Impact of PEGylated ligands on lung function and inflammatory changes was studied in animal models of acute lung injury, asthma, and silicosis following Lipopolysaccharide (LPS), allergen (ovalbumin), and silica inhalation, respectively. Subcutaneous injection of TMX- 302 prevented LPS- and allergen-induced airway hyper-reactivity (AHR), leukocyte infiltration, and production of pro-inflammatory cytokines in the lung. However, intranasal instillation of TMX-302 led to neutrophil infiltration and failed to prevent allergen-induced AHR, despite inhibiting leukocyte counts in the BAL. Aerosolized TMX-306 given prophylactically, but not therapeutically, inhibited pivotal asthma features. Interventional treatment with intranasal instillation of TMX-306 significantly reduced the pulmonary fibrogranulomatous response and the number of silica particles in lung interstitial space in silicotic mice. These findings highlight the potential of TMX-306, emphasizing its value in drug development for lung diseases, and particularly silicosis

Putative protective effect of Cadmium chloride high diluted solution on LLC-PK1 cell intoxicated by high concentration of this same metal: an isopathic in vitro assay

ABSTRACT Cadmium is an important toxic environmental heavy metal. Several studies have demonstrated that a major site of cadmium toxicity in humans and in other animals is the proximal tubule of the kidney. A well established model for nefrotoxicity is the use of in vitro technique with proximal tubule epithelial cell lines, as LLC-PK1. Herein, we have the intention to study the possible protective effect of high diluted CdCl2 solutions. In a blinding way, LLC-PK1 cells were pre-treated with high diluted cadmium chloride in the potencies 10 cH, 15 cH and 20cH. After 4 days, these cells have received CdCl2 in a pre-determined toxic concentration. The cell viability was assessed by MTT assay. We have identified a protective effect of two CdCl2 high diluted solutions, 10 cH and 20 cH, when cells were intoxicated by sublethal CdCl2 concentration. The results indicate that probably the high dilutions have an expressive action on cells in sublethal intoxication

Aplicação de anticorpos biespecíficos como imunoreagentes : estudo da cinética de reação da enzima fosfatase alcalina e sua modulação com anticorpos anti-enzima

06

Avaliação de afinidade e especificidade de um anticorpo monoclonal anti-insulina : ensaios preliminares para desenvolvimento de radioimunoensaio

07

Determinação cinética de reação da enzima peroxidase e sua modulação com anticorpos anti-enzima : implicações no desenvolvimento de anticorpos biespecíficos

21

Potential of PEGylated Toll-Like Receptor 7 Ligands for Controlling Inflammation and Functional Changes in Mouse Models of Asthma and Silicosis

Submitted by Sandra Infurna ([email protected]) on 2016-12-27T12:47:07Z No. of bitstreams: 1 tatiana_ferreira_etal_IOC_2016.pdf: 8576080 bytes, checksum: c9648fefb16d2011b1b0f04f2bd13400 (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2016-12-27T12:57:38Z (GMT) No. of bitstreams: 1 tatiana_ferreira_etal_IOC_2016.pdf: 8576080 bytes, checksum: c9648fefb16d2011b1b0f04f2bd13400 (MD5)Made available in DSpace on 2016-12-27T12:57:38Z (GMT). No. of bitstreams: 1 tatiana_ferreira_etal_IOC_2016.pdf: 8576080 bytes, checksum: c9648fefb16d2011b1b0f04f2bd13400 (MD5) Previous issue date: 2016Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Inflamação. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Inflamação. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Inflamação. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Inflamação. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Inflamação. Rio de Janeiro, RJ, Brasil.Universitá della Svizzera Italiana. Bellnzona, Switzerland.Universitá della Svizzera Italiana. Bellnzona, Switzerland.Universitá della Svizzera Italiana. Bellnzona, Switzerland.Telormedix SA. Bioggio, Switzerland.Telormedix SA. Bioggio, Switzerland.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Inflamação. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Inflamação. Rio de Janeiro, RJ, Brasil.Prior investigations show that signaling activation through pattern recognition receptors can directly impact a number of inflammatory lung diseases. While toll-like receptor (TLR) 7 agonists have raised interest for their ability to inhibit allergen-induced pathological changes in experimental asthma conditions, the putative benefit of this treatment is limited by adverse effects. Our aim was to evaluate the therapeutic potential of two PEGylated purine-like compounds, TMX-302 and TMX-306, characterized by TLR7 partial agonistic activity; therefore, the compounds are expected to induce lower local and systemic adverse reactions. In vitro approaches and translation to murine models of obstructive and restrictive lung diseases were explored. In vitro studies with human PBMCs showed that both TMX-302 and TMX-306 marginally affects cytokine production as compared with equivalent concentrations of the TLR7 full agonist, TMX-202. The PEGylated compounds did not induce monocyte-derived DC maturation or B cell proliferation, differently from what observed after stimulation with TMX-202. Impact of PEGylated ligands on lung function and inflammatory changes was studied in animal models of acute lung injury, asthma, and silicosis following Lipopolysaccharide (LPS), allergen (ovalbumin), and silica inhalation, respectively. Subcutaneous injection of TMX-302 prevented LPS- and allergen-induced airway hyper-reactivity (AHR), leukocyte infiltration, and production of pro-inflammatory cytokines in the lung. However, intranasal instillation of TMX-302 led to neutrophil infiltration and failed to prevent allergen-induced AHR, despite inhibiting leukocyte counts in the BAL. Aerosolized TMX-306 given prophylactically, but not therapeutically, inhibited pivotal asthma features. Interventional treatment with intranasal instillation of TMX-306 significantly reduced the pulmonary fibrogranulomatous response and the number of silica particles in lung interstitial space in silicotic mice. These findings highlight the potential of TMX-306, emphasizing its value in drug development for lung diseases, and particularly silicosis