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1.
J Phys Chem Lett ; 10(20): 6105-6111, 2019 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-31549842

RESUMO

Protein-protein interactions (PPIs) play a pivotal role in many biological processes. Discriminating functionally important well-defined protein-protein complexes formed by specific interactions from random aggregates produced by nonspecific interactions is therefore a critical capability. While there are many techniques which enable rapid screening of binding affinities in PPIs, there is no generic spectroscopic phenomenon which provides rapid characterization of the structure of protein-protein complexes. In this study we show that chiral plasmonic fields probe the structural order and hence the level of PPI specificity in a model antibody-antigen system. Using surface-immobilized Fab' fragments of polyclonal rabbit IgG antibodies with high specificity for bovine serum albumin (BSA), we show that chiral plasmonic fields can discriminate between a structurally anisotropic ensemble of BSA-Fab' complexes and random ovalbumin (OVA)-Fab' aggregates, demonstrating their potential as the basis of a useful proteomic technology for the initial rapid high-throughput screening of PPIs.


Assuntos
Fragmentos Fab das Imunoglobulinas/metabolismo , Imunoglobulina G/metabolismo , Nanoestruturas/química , Cimento de Policarboxilato/química , Soroalbumina Bovina/metabolismo , Animais , Anisotropia , Bovinos , Ouro/química , Fragmentos Fab das Imunoglobulinas/imunologia , Imunoglobulina G/imunologia , Ovalbumina/imunologia , Ovalbumina/metabolismo , Ligação Proteica , Coelhos , Soroalbumina Bovina/imunologia , Análise Espectral/métodos , Estereoisomerismo
2.
Exp Parasitol ; 206: 107767, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31520603

RESUMO

Schistosoma mansoni eggs can influence immune responses directed at them, and the mechanisms by which this is achieved are being unravelled. Going towards, developing effective tools for the study of how S. mansoni influences naïve T cells, we have developed S. mansoni eggs expressing chicken ovalbumin (OVA), using a lentiviral transduction system. Indeed, such a parasite could be used in conjunction with cells from OT-II transgenic mice as a source of naïve, antigen-specific T cells. The expression of the transgenic protein was confirmed by real-time RT-PCR of OVA-specific mRNA and western blotting using polyclonal antibodies specific for OVA. T cells from OT-II transgenic mice expressing a T cell receptor specific for the OVA323-339 peptide recognised the OVA-transduced S. mansoni eggs. Using flow cytometry on CFSE-labelled OT-II splenocytes, we demonstrated that OVA-transduced eggs elicit higher OT-II proliferative responses than untransduced eggs. The OT-II T cells also produced TNF-α and IFN-γ following exposure to OVA-transduced eggs. In addition, moderate amounts of IL-6 and IL-17A were also detected. In contrast, no IL-10, IL-4 and IL-2 were detected in cultures, whether the cells were stimulated with transduced or untransduced eggs. Thus, the cytokine signatures showed the transfected eggs induced predominantly a Th1 response, with a small amount of IL-6 and IL-17.


Assuntos
Ovalbumina/análise , Receptores de Antígenos de Linfócitos T/imunologia , Schistosoma mansoni/metabolismo , Linfócitos T/imunologia , Animais , Western Blotting , Galinhas , Citocinas/análise , Citocinas/metabolismo , Eletroforese em Gel de Ágar , Feminino , Citometria de Fluxo , Interleucina-17/análise , Interleucina-17/metabolismo , Interleucina-2/análise , Interleucina-2/metabolismo , Interleucina-6/análise , Interleucina-6/metabolismo , Fígado/parasitologia , Linfonodos/citologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Transgênicos , Ovalbumina/genética , Ovalbumina/imunologia , Ovalbumina/metabolismo , Óvulo/metabolismo , RNA Mensageiro/análise , RNA Mensageiro/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Antígenos de Linfócitos T/genética , Transcrição Reversa , Schistosoma mansoni/genética , Schistosoma mansoni/crescimento & desenvolvimento , Baço/citologia , Linfócitos T/citologia
3.
Int J Nanomedicine ; 14: 5229-5242, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31371958

RESUMO

Purpose: Dexamethasone (Dex) has long been used as a potent immunosuppressive agent in the treatment of inflammatory and autoimmune diseases, despite serious side effects. In the present study, Dex and model antigen ovalbumin (OVA) were encapsulated with poly(lactic-co-glycolic acid) to deliver Dex and OVA preferentially to phagocytic cells, reducing systemic side effects of Dex. The OVA-specific immune tolerance-inducing activity of the nanoparticles (NPs) was examined. Methods: Polymeric NPs containing OVA and Dex (NP[OVA+Dex]) were prepared by the water-in-oil-in-water double emulsion solvent evaporation method. The effects of NP[OVA+Dex] on the maturation and function of immature dendritic cells (DCs) were examined in vitro. Furthermore, the OVA-specific immune tolerizing effects of NP[OVA+Dex] were confirmed in mice that were intravenously injected or orally fed with the NPs. Results: Immature DCs treated in vitro with NP[OVA+Dex] did not mature into immunogenic DCs but instead were converted into tolerogenic DCs. Furthermore, profoundly suppressed generation of OVA-specific cytotoxic T cells and production of OVA-specific IgG were observed in mice injected with NP[OVA+Dex], whereas regulatory T cells were concomitantly increased. Feeding of mice with NP[OVA+Dex] also induced OVA-specific immune tolerance. Conclusion: The present study demonstrates that oral feeding as well as intravenous injection of poly(lactic-co-glycolic acid) NPs encapsulating both antigen and Dex is a useful means of inducing antigen-specific immune tolerance, which is crucial for the treatment of autoimmune diseases.


Assuntos
Antígenos/imunologia , Materiais Biocompatíveis/química , Dexametasona/farmacologia , Epitopos/imunologia , Tolerância Imunológica , Nanopartículas/química , Administração Oral , Animais , Apresentação do Antígeno/efeitos dos fármacos , Citocinas/biossíntese , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Feminino , Fatores de Transcrição Forkhead/metabolismo , Imunidade/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Nanopartículas/administração & dosagem , Ovalbumina/imunologia , Fagocitose/efeitos dos fármacos , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Linfócitos T Citotóxicos/imunologia , Linfócitos T Reguladores/efeitos dos fármacos , Linfócitos T Reguladores/metabolismo
4.
Environ Pollut ; 252(Pt B): 1519-1531, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31277021

RESUMO

Some basic research has shown that nanomaterials can aggravate allergic asthma. However, its potential mechanism is insufficient. Based on the research that alumina nanopowder (nAl2O3) has been reported to cause lung tissue damage, the purpose of this study was to explore the relationship between nAl2O3 and allergic asthma as well as its molecular mechanism. In this study, Balb/c mice were sensitized with ovalbumin (OVA) to construct the allergic asthma model while intratracheally administered 0.5, 5 or 50 mg kg-1·day-1 nAl2O3 for 3 weeks. It was observed that exposure to nAl2O3 exacerbated airway hyperresponsiveness (AHR), airway remodeling, and inflammation cell infiltration, leading to lung function damage in mice. Results revealed that nAl2O3 could increase ROS levels and decrease GSH levels in lung tissue, promote the increases of the T-IgE, TGF-ß, IL-1ß and IL-6 levels, stimulate the overexpression of transcription factors GATA-3 and RORγt, decrease the levels of IFN-γ and IL-10 and increase the levels of IL-4 and IL-17A, resulting in the imbalance of Th1/Th2 and Treg/Th17 immune responses. In addition, antioxidant Vitamin E (Vit E) could alleviate asthma-like symptoms through blocking oxidative stress. The study displayed that exposure of nAl2O3 deteriorated allergic asthma through promoting the imbalances of Th1/Th2 and Treg/Th17.


Assuntos
Antioxidantes/farmacologia , Asma/prevenção & controle , Estresse Oxidativo/efeitos dos fármacos , Células Th17/efeitos dos fármacos , Células Th2/efeitos dos fármacos , Vitamina E/farmacologia , Óxido de Alumínio/toxicidade , Animais , Asma/induzido quimicamente , Asma/imunologia , Interleucina-17/imunologia , Interleucina-4/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Nanopartículas/toxicidade , Ovalbumina/imunologia , Estresse Oxidativo/imunologia , Linfócitos T Reguladores/imunologia , Células Th17/imunologia , Células Th2/imunologia
5.
J Agric Food Chem ; 67(29): 8138-8148, 2019 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-31294563

RESUMO

The aim of the present study was to compare various glycated ovalbumin (OVA)-monosaccharides, including OVA-mannose (Man), -glucose, -ribose, and -fructose, in the attenuation of OVA-induced allergic response in a BALB/C mouse model and the potential mechanisms of immunological modulation. The glycated OVA forms were prepared by Maillard reactions. OVA-Man significantly reduced the frequency of allergic signs. Mouse mast cell protease enzyme concentration was significantly reduced in the OVA-Man group (549.80 ± 84.67 ng/mL, p < 0.05). The OVA-Man group also had a lower histamine concentration (30.96 ± 1.12 ng/mL) as compared with the positive control OVA group (44.43 ± 0.71 ng/mL, p < 0.05). Both specific IgG and IgE were significantly reduced in the OVA-Man-treated group (p < 0.05). The OVA-Man group exhibited decreased concentrations of IL-4 (67.98 ± 3.11 pg/mL) and IL-17 (67.98 ± 3.11 pg/mL) and an increased concentration of IL-12 (336.70 ± 18.69 pg/mL, p < 0.05) compared with the positive control. Mannosylation played a vital role in allergen recognition, implicating deleterious downstream Th2 cell activation, cytokine secretion, and IgE production. This result indicates that different glycans target specific DC receptors, and differential DC processing, antigen presentation, and T cell response leads to altered variation in allergic response. OVA-Man exhibited minimal DC internalization, DC processing, MHC antigen presentation, and antigen-specific T cell activation, resulting in an attenuated allergic response and validating its efficacy as a potential immunotherapeutic candidate to treat egg allergy.


Assuntos
Hipersensibilidade a Ovo/imunologia , Monossacarídeos/química , Ovalbumina/química , Ovalbumina/imunologia , Animais , Citocinas/imunologia , Células Dendríticas/imunologia , Modelos Animais de Doenças , Hipersensibilidade a Ovo/etiologia , Feminino , Glicosilação , Humanos , Imunoglobulina E/imunologia , Reação de Maillard , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/efeitos adversos , Linfócitos T/imunologia
6.
Nat Med ; 25(7): 1164-1174, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31235962

RESUMO

The role of dysbiosis in food allergy (FA) remains unclear. We found that dysbiotic fecal microbiota in FA infants evolved compositionally over time and failed to protect against FA in mice. Infants and mice with FA had decreased IgA and increased IgE binding to fecal bacteria, indicative of a broader breakdown of oral tolerance than hitherto appreciated. Therapy with Clostridiales species impacted by dysbiosis, either as a consortium or as monotherapy with Subdoligranulum variabile, suppressed FA in mice as did a separate immunomodulatory Bacteroidales consortium. Bacteriotherapy induced expression by regulatory T (Treg) cells of the transcription factor ROR-γt in a MyD88-dependent manner, which was deficient in FA infants and mice and ineffectively induced by their microbiota. Deletion of Myd88 or Rorc in Treg cells abrogated protection by bacteriotherapy. Thus, commensals activate a MyD88/ROR-γt pathway in nascent Treg cells to protect against FA, while dysbiosis impairs this regulatory response to promote disease.


Assuntos
Hipersensibilidade Alimentar/terapia , Microbioma Gastrointestinal/imunologia , Fator 88 de Diferenciação Mieloide/fisiologia , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/fisiologia , Linfócitos T Reguladores/fisiologia , Animais , Bacteroides , Clostridiales , Disbiose/imunologia , Fezes/microbiologia , Hipersensibilidade Alimentar/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Ovalbumina/imunologia , Transdução de Sinais
7.
Iran J Allergy Asthma Immunol ; 18(2): 209-217, 2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-31066257

RESUMO

Asthma is a chronic inflammatory disease of the airways of the lungs. Pomalidomide (POM) a therapy for multiple myeloma has been stated to have an anti-inflammatory effect. The main goal of the present study was to assess its possible effect on airway contraction and inflammation in a rat model of ovalbumin-induced asthma. Different groups of rats received saline or pomalidomide (0.4, 0.8 mg/kg) or dexamethasone (0.6 mg/kg). The asthma was induced by ovalbumin (OVA). Trachea contraction was assayed by organ bath system. Airway histology was assessed using hematoxylin and eosin method. Serum Tumor necrosis factor alpha (TNF-α) level was analyzed by Enzyme-Linked Immunosorbent Assay and Platelet-derived growth factor (PDGFα) Gene expressions were evaluated by Real-time PCR. Pomalidomide prevented ovalbumin-induced airway contraction and histopathological damage. In addition serum, TNF-α level was significantly (p<0.05) decreased in POM treated animals compared to control (asthmatic animals that received POM vehicle). Results indicate that POM prevented the PDGF expression induced by ovalbumin. In conclusion, we found that pomalidomide ameliorated the symptoms, histopathological changes and inflammatory markers induced by ovalbumin in asthmatic rats and these effects might be related to its anti-inflammatory properties.


Assuntos
Obstrução das Vias Respiratórias/tratamento farmacológico , Alérgenos/imunologia , Anti-Inflamatórios/uso terapêutico , Asma/tratamento farmacológico , Pulmão/patologia , Talidomida/análogos & derivados , Animais , Modelos Animais de Doenças , Humanos , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Masculino , Ovalbumina/imunologia , Fator de Crescimento Derivado de Plaquetas/metabolismo , Ratos , Ratos Wistar , Talidomida/uso terapêutico , Fator de Necrose Tumoral alfa/sangue
8.
Int J Nanomedicine ; 14: 3221-3234, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31123399

RESUMO

Background: Poly (lactic-co-glycolic acid) (PLGA) nanoparticles and surface modified PLGA nanoparticles have been widely studied as antigens or drugs carriers due to their controlled release characteristics and biocompatibility. However, most PLGA nanoparticles have lower antigens loading efficiency and adjuvanticity. Purpose: The aim of this study was to improve the antigen loading efficiency and adjuvant activity of PLGA nanoparticles. Materials and methods: Surface cationic polymer modification can improve the antigens loading efficiency of PLGA nanoparticles by surface adsorption. Therefore, in this study, chitosan modified PLGA nanoparticles (CS-AHPP/OVA), polyethyleneimine modified PLGA nanoparticles (PEI-AHPP/OVA), and ε-Poly-L-lysine modified PLGA nanoparticles (εPL-AHPP/OVA) were prepared as antigen delivery carriers to investigate the characterization and stability of these nanoparticles. These nanoparticles were evaluated for their efficacies as adjuvants pre- and post-modification. Results: The AHP and OVA-loaded PLGA nanoparticles (AHPP/OVA) were positively charged after surface cationic polymers modification, and their structural integrity was maintained. Their antigen loading capacity and stability of nanoparticles were improved by the surface cationic polymers modification. Increased positive surface charge resulted in greater OVA adsorption capacity. Among AHPP/OVA and the three surface cationic polymers synthesized from modified PLGA nanoparticles, PEI-AHPP/OVA showed the highest antigen loading efficiency and good stability. AHPP/OVA, CS-AHPP/OVA PEI-AHPP/OVA, and εPL-AHPP/OVA formulations significantly enhanced lymphocyte proliferation and improved the ratio of CD4+/CD8+ T cells. In addition, AHPP/OVA, PEI-AHPP/OVA and εPL-AHPP/OVA formulations induced secretion of cytokines (TNF-α, IFN-γ, IL-4, and IL-6), antibodies (IgG) and antibody subtypes (IgG1 and IgG2a) in immunized mice. These results demonstrate that these formulations generated a strong Th1-biased immune response. Among them, PEI-AHPP/OVA induced the strongest Th1-biased immune response. Conclusion: In conclusion, PEI-AHPP/OVA nanoparticles may be a potential antigen delivery system for the induction of strong immune responses.


Assuntos
Sistemas de Liberação de Medicamentos , Mel , Imunidade Celular , Ovalbumina/administração & dosagem , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Polissacarídeos/química , Vacinas/administração & dosagem , Adjuvantes Imunológicos/administração & dosagem , Animais , Formação de Anticorpos , Antígenos/metabolismo , Cátions , Proliferação de Células , Quitosana/química , Citocinas/sangue , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Feminino , Humanos , Imunidade Celular/efeitos dos fármacos , Ativação Linfocitária , Camundongos Endogâmicos ICR , Nanopartículas/química , Ovalbumina/imunologia , Polietilenoimina/química , Baço/citologia , Linfócitos T/imunologia , Vacinação , Vacinas/imunologia
9.
Eur J Pharm Biopharm ; 140: 29-39, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31055066

RESUMO

Using subunit vaccines, e.g., based on peptide or protein antigens, to teach the immune system to kill abnormal host cells via induction of cytotoxic T lymphocytes (CTL) is a promising strategy against intracellular infections and cancer. However, customized adjuvants are required to potentiate antigen-specific cellular immunity. One strong CTL-inducing adjuvant is the liposomal cationic adjuvant formulation (CAF)09, which is composed of dimethyldioctadecylammonium (DDA) bromide, monomycoloyl glycerol (MMG) analogue 1 and polyinosinic:polycytidylic acid [poly(I:C)]. However, this strong CTL induction requires intraperitoneal administration because the vaccine forms a depot at the site of injection (SOI) after subcutaneous (s.c.) or intramuscular (i.m.) injection, and depot formation impedes the crucial vaccine targeting to the cross-presenting dendritic cells (DCs) residing in the lymph nodes (LNs). The purpose of the present study was to investigate the effect of polyethylene glycol (PEG) grafting of CAF09 on the ability of the vaccine to induce antigen-specific CTL responses after s.c. administration. We hypothesized that steric stabilization and charge shielding of CAF09 by PEGylation may reduce depot formation at the SOI and enhance passive drainage to the LNs, eventually improving CTL induction. Hence, the vaccine (antigen/CAF09) was post-grafted with a novel type of anionic PEGylated peptides based on GDGDY repeats, which were end-conjugated with one or two PEG1000 moieties, resulting in mono- and bis-PEG-peptides of different lengths (10, 15 and 20 amino acid residues). For comparison, CAF09 was also grafted by inclusion of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-methoxy(PEG)-2000 (DSPE-PEG2000) in the bilayer structure during preparation. Grafting of CAF09 with either type of PEG resulted in charge shielding, evident from a reduced surface charge. Upon s.c. immunization of mice with the model antigen ovalbumin (OVA) adjuvanted with PEGylated CAF09, stronger CTL responses were induced as compared to immunization of mice with unadjuvanted OVA. Biodistribution studies confirmed that grafting of CAF09 with DSPE-PEG2000 improved the passive drainage of the vaccine to LNs, because a higher dose fraction was recovered in DCs present in the draining LNs, as compared to the dose fraction detected for non-PEGylated CAF09. In conclusion, PEGylation of CAF09 may be a useful strategy for the design of an adjuvant, which induces CTL responses after s.c. and i.m. administration. In the present studies, CAF09 grafted with 10 mol% DSPE-PEG2000 is the most promising of the tested adjuvants, but additional studies are required to further elucidate the potential of the strategy.


Assuntos
Adjuvantes Imunológicos/química , Adjuvantes Imunológicos/farmacologia , Lipossomos/química , Polietilenoglicóis/química , Linfócitos T Citotóxicos/imunologia , Vacinas de Subunidades/imunologia , Animais , Antígenos/imunologia , Linfócitos T CD8-Positivos/imunologia , Apresentação Cruzada/imunologia , Células Dendríticas/imunologia , Feminino , Imunidade Celular/imunologia , Imunização/métodos , Camundongos , Camundongos Endogâmicos C57BL , Ovalbumina/imunologia , Fosfatidiletanolaminas/química , Compostos de Amônio Quaternário/química , Distribuição Tecidual
10.
Int Arch Allergy Immunol ; 179(3): 192-200, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30999298

RESUMO

AIM: Asthma appears to be a common comorbid condition in children with sickle cell disease (SCD), and such individuals may be at a higher risk for increased morbidity and mortality. However, several reports have indicated that asthma severity is not particularly high in those with SCD, and airway hyperreactivity and wheeze may be independently associated with SCD. In SCD mice, exacerbated allergic airway disease (AAD) has been observed in response to the model antigen ovalbumin (OVA). We sought to determine if allergic lung inflammation is also exacerbated in SCD mice when they are exposed to the human allergen, house dust mite (HDM). METHODS AND RESULTS: Eosinophil counts in bronchoalveolar lavage fluid were determined by cytocentrifugation and increased in both wild-type (WT) and SCD mice after acute exposure to a high dose (25 µg) of HDM, which then decreased in chronically exposed mice. WT mice exposed to a low dose of HDM (1 µg) followed the same pattern of eosinophil flux, but SCD mice did not induce much eosinophilia after acute exposure to HDM. As was observed in previous studies, lung lesions similarly increased in severity in both WT and SCD mice after acute exposure to HDM, which remained elevated after chronic exposure. Furthermore, serum HDM-specific IgE titers similarly increased and selected serum cytokines were similar in both WT and SCD mice. CONCLUSION: These results contrast with previous reports of exacerbated AAD in SCD mice exposed to OVA and support the alternative hypothesis that asthmatic responses are normal in those with SCD.


Assuntos
Alérgenos/imunologia , Anemia Falciforme/imunologia , Ovalbumina/imunologia , Pyroglyphidae/imunologia , Hipersensibilidade Respiratória/imunologia , Anemia Falciforme/sangue , Animais , Líquido da Lavagem Broncoalveolar/imunologia , Citocinas/sangue , Feminino , Imunoglobulina E/sangue , Contagem de Leucócitos , Masculino , Camundongos Transgênicos , Hipersensibilidade Respiratória/sangue
11.
Int J Mol Sci ; 20(8)2019 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-30991656

RESUMO

Dipsacus asperoides C. Y. Cheng et T. M. Ai (DA) has been used in China as a traditional medicine to treat lumbar and knee pain, liver dysfunction, and fractures. We explored the suppressive effect of DA on allergic asthma using an ovalbumin (OVA)-induced asthma model. In the asthma model, female Balb/c mice were sensitized to OVA on day 0 and 14 to boost immune responses and then exposed to OVA solution by using an ultrasonic nebulizer on days 21 to 23. DA (20 and 40 mg/kg) was administered to mice by oral gavage on days 18 to 23. Methacholine responsiveness was determined on day 24 using a plethysmography. On day 25, we collected bronchoalveolar lavage fluid, serum, and lung tissue from animals under anesthesia. DA treatment effectively inhibited methacholine responsiveness, inflammatory cell infiltration, proinflammatory cytokines such as interleukin (IL)-5 and IL-13, and immunoglobulin (Ig) E in OVA-induced asthma model. Reductions in airway inflammation and mucus hypersecretion, accompanied by decreases in the expression of inducible nitric oxide synthase (iNOS) and the phosphorylation of nuclear factor kappa B (NF-κB), were also observed. Our results indicated that DA attenuated the asthmatic response, and that this attenuation was closely linked to NF-κB suppression. Thus, this study suggests that DA is a potential therapeutic for allergic asthma.


Assuntos
Anti-Inflamatórios/uso terapêutico , Asma/tratamento farmacológico , Dipsacaceae , Medicamentos de Ervas Chinesas/uso terapêutico , Animais , Anti-Inflamatórios/química , Asma/etiologia , Asma/imunologia , Dipsacaceae/química , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas/química , Feminino , Imunoglobulina E/imunologia , Interleucina-13/imunologia , Interleucina-5/imunologia , Camundongos Endogâmicos BALB C , NF-kappa B/imunologia , Ovalbumina/imunologia
12.
Methods Mol Biol ; 1957: 335-343, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30919364

RESUMO

Spatial and temporal control of gene expression using cre/loxP technology is a major methodological advance for biomedical research. The ability to alter gene expression after an in vivo disease model has been established and allows researchers the opportunity to examine the impact of that gene on the perpetuation of a disease, a mechanistic insight that is arguably more therapeutically relevant than developmental mechanisms.We used the cre/LoxP technology in mice to show that ß-arrestin-2, a gene previously shown to be required for the development of the asthma phenotype, is also required for the perpetuation of, at least, the airway hyperresponsiveness characteristic of that phenotype. Here we describe stepwise methods for the activation of the cre-loxP technology and induction of murine allergic inflammatory airway disease. We comment on the unanticipated problems encountered, which we speculate were a result of interactions between the allergen and ß-arrestin-2 gene (Arrb2) regulation and the effect of tamoxifen on the asthma phenotype. The issues encountered here may be generally applicable to cre/loxP utilization in inflammatory disease models, especially if the gene of interest is associated with the inflammatory cascade.


Assuntos
Hipersensibilidade/metabolismo , Inflamação/metabolismo , Biologia Molecular/métodos , beta-Arrestina 2/metabolismo , Animais , Asma/imunologia , Modelos Animais de Doenças , Camundongos Knockout , Ovalbumina/imunologia
13.
Pharmacol Rep ; 71(2): 330-337, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30826574

RESUMO

BACKGROUND: Food allergy (FA) is a worldwide health problem, affecting nearly 10% of all populations, with no prophylactic options or regulatory treatment available until now. Fisetin, a biologically active flavonoid, and telmisartan, the highly selective competitive AT1 receptor antagonist, recently exhibited potent anti-inflammatory and immunomodulatory activities. In the present study, we have evaluated the possible anti-inflammatory and immunomodulatory activities of fisetin and telmisartan each alone or in low-dose combination in a mouse model of FA. METHODS: For induction of FA, eight-week-old BALB/c mice, sensitized by two ip injection of 50 µg ovalbumin (OVA) and 1 mg alum at day 0 and 7. Then, each mouse challenged with 10 mg OVA at days 14, 16, 18, and 21. On the 28th day, the fifth challenge carried out by oral administration of 50 mg OVA. Either fisetin (1 or 3 mg/kg/d), telmisartan (1 or 3 mg/kg/d) or a combination of fisetin 1 mg/kg/d and telmisartan 1 mg/kg/d received orally from the 13th day till 28th day. In challenge days, the treatments received one-hour before the challenge. RESULTS: Our data showed that fisetin and telmisartan each alone or in low-dose combination attenuated the anaphylactic manifestation, decreased blood eosinophilic count, serum OVA-specific IgE, and IL-4 levels, the intestinal total and degranulated mast cells count, and CD4+ immunohistochemical expression. Furthermore, they enhanced the serum IFN-γ level and abrogated the intestinal histopathological changes induced by OVA in mice. CONCLUSION: Either fisetin, telmisartan or their low-dose combination could be promising in the management of FA.


Assuntos
Flavonoides/farmacologia , Hipersensibilidade Alimentar/tratamento farmacológico , Fatores Imunológicos/farmacologia , Telmisartan/farmacologia , Animais , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Quimioterapia Combinada , Flavonoides/administração & dosagem , Hipersensibilidade Alimentar/imunologia , Fatores Imunológicos/administração & dosagem , Interferon gama/sangue , Intestinos/imunologia , Mastócitos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Telmisartan/administração & dosagem , Fatores de Tempo
14.
Biomolecules ; 9(3)2019 03 13.
Artigo em Inglês | MEDLINE | ID: mdl-30871269

RESUMO

Anaphylactic shock (AS) is a life-threatening, multisystem disorder arising from sudden release of mast cell- and basophil-derived mediators into the circulation. In this study, we have used a Wistar rat model to investigate AS-associated histopathologic changes in various organs. Rats were sensitized with ovalbumin (1 mg s.c), and AS was induced by intravenous injection of ovalbumin (1 mg). Experimental groups included nonallergic rats (n = 6) and allergic rats (n = 6). Heart rate and blood pressure were monitored during one hour. Organs were harvested at the end of the experiment and prepared for histologic and immunohistochemical studies. Lung, small bowel mucosa and spleen were found to undergo heavy infiltration by mast cells and eosinophils, with less prominent mast cell infiltration of cardiac tissue. The mast cells in lung, small bowel and spleen exhibited increased expression of tryptase, c-kit and induced nitric oxide synthase (iNOS). Increased expression of endothelial nitric oxide synthase (eNOS) by vascular endothelial cells was noted principally in lung, heart and small bowel wall. The Wistar rat model of AS exhibited accumulation of mast cells and eosinophils in the lung, small bowel, and spleen to a greater extent than in the heart. We conclude that lung and gut are principal inflammatory targets in AS, and likely contribute to the severe hypotension of AS. Targeting nitric oxide (NO) production may help reduce AS mortality.


Assuntos
Anafilaxia/imunologia , Anafilaxia/patologia , Hipotensão/patologia , Inflamação/patologia , Ovalbumina/imunologia , Animais , Modelos Animais de Doenças , Hipotensão/imunologia , Inflamação/imunologia , Injeções Intravenosas , Injeções Subcutâneas , Masculino , Óxido Nítrico/biossíntese , Ovalbumina/administração & dosagem , Ratos , Ratos Wistar
15.
Mol Med Rep ; 19(4): 3210-3216, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30816463

RESUMO

Asthma is a chronic lung disease characterized by an imbalance of T­helper (Th)1/Th2 cells and their cytokine profiles. Natural killer (NK) cells constitute a considerable subset of the lymphocyte population in the lungs, and provide protection against respiratory infection by fungi, bacteria and viruses. However, the mechanism by which NK cells are involved in asthma remains to be fully elucidated. The present study analyzed the dynamic changes of NK cells and their subsets during the development of the ovalbumin (OVA)­induced allergic airway response. Lung tissues were histologically examined for cell infiltration and mucus hypersecretion. The number, activity and cytokine­secreting ability of NK cells was determined by flow cytometry. The results showed that the percentage of NK cells in the lung was decreased following OVA sensitization and challenge. However, NK cells exhibited enhanced activity and secreted more Th2 cytokines (IL­5 and IL­13) following OVA challenge. Furthermore, the proportion of CD11b­ NK subsets increased with the development of asthma, and CD11b­ CD27­ NK cells were the primary NK subset producing Th2 cytokines. These findings suggest that, although NK cells are not the crucial type of lymphocytes involved in asthma, OVA induces NK cells to secrete Th2 cytokines that may be involved in the pathogenesis of asthma.


Assuntos
Asma/imunologia , Asma/metabolismo , Interleucina-13/metabolismo , Interleucina-5/metabolismo , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Ovalbumina/imunologia , Alérgenos/imunologia , Animais , Biomarcadores , Modelos Animais de Doenças , Feminino , Imunofenotipagem , Camundongos , Células Th2/imunologia , Células Th2/metabolismo
16.
J Ethnopharmacol ; 236: 82-90, 2019 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-30836174

RESUMO

OBJECTIVE: San'ao decoction (SAD), a traditional Chinese prescription, is well-known in asthma treatment. In the current study, the protective role of SAD and its mechanism in aggravated asthma mice model via regulation of TRP channel were evaluated and explored. METHODS: UPLC-QTOF-MS was used for analyzing the chemicals in SAD. The major chemical components in SAD were separated and detected under an optimized chromatographic and MS condition. 75 BALB/c mice were randomly divided into five groups: normal group, model group, dexamethasone group (0.75 mg kg-1), SAD-high dose group (1.8 g kg-1) and SAD-low dose group (0.9 g kg-1). A 42 days aggravated asthmatic model was established in mice induced by ovalbumin (OVA) plus PM2.5 (1.6 mg kg-1). After treated with corresponding medicine, peripheral blood and bronchoalveolar lavage fluid (BALF) from each group were assessed, airway responsiveness was determined, histopathological changes in lungs were detected, relevant cytokines and neurokines levels were measured, TRPA1 and TRPV1 mRNA and protein expressions in lung tissues were examined as well. RESULTS: 21 signal peaks of the chemicals in SAD were identified with the method of UPLC-QTOF-MS. SAD, especially SAD-high dose exerted significant effects on OVA plus PM2.5 mice model in relieving lung injury score (P < 0.05), reducing eosinophil (EOS) count in blood (P < 0.05) and inflammatory cells ratio in BALF (P < 0.05, P < 0.01), decreasing RI value (P < 0.05) while increasing Cdyn value (P < 0.05), reducing IL-13, PGD2 and NGF levels in BALF (P < 0.01), as well as down-regulating TRPA1 and TRPV1 mRNA and protein expressions in lung tissues (P < 0.05, P < 0.01). CONCLUSION: SAD could improve pulmonary functions, relieve lung injury, as well as reduce IL-13, PGD2 and NGF levels of OVA plus PM2.5 aggravated asthma model in mice. The effect and mechanism of SAD might be related to the inhibition of TRPA1 and TRPV1 channels.


Assuntos
Asma/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Material Particulado/toxicidade , Canal de Cátion TRPA1/metabolismo , Canais de Cátion TRPV/metabolismo , Animais , Asma/induzido quimicamente , Asma/imunologia , Líquido da Lavagem Broncoalveolar/citologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Lesão Pulmonar/prevenção & controle , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Testes de Função Respiratória
17.
Am J Rhinol Allergy ; 33(2): 145-152, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30871345

RESUMO

BACKGROUND: Predominantly, 2 animal models are used for allergic rhinitis (AR), which are established by intraperitoneal (IP) injection plus local challenge and nasal-only delivery. The differences between these 2 models are not fully understood. Moreover, dose-response relationship to allergens remains unclear. METHODS: In this study, mice were sensitized by nasal drops (without adjuvant, once daily for 9 weeks) to set up a nasal-only delivery AR model. Five different doses of ovalbumin (OVA) nasal drops were served to explore the dose-response to allergens. Allergic symptoms, serum antibodies (IgE, IgG2a, and IgG1), spleen supernatant and nasal lavage fluid (NALF) cytokines (IL-4, IL-5, and IFN-r), and infiltrated eosinophils of the nasal mucosa were observed. RESULTS: The allergic symptoms, serum antibodies, cytokines, and infiltrated eosinophils were significantly higher in the high OVA concentration compared with those of the control group. Different OVA concentrations associated with the severity of allergy. Within a certain concentration range, OVA concentration positively related to the severity of symptoms, IgE antibody level, and Th2 bias. Meanwhile, serum antibodies (IgE and IgG1) and cytokines (IL-4, IL-5 in spleen and IL-4 in NALF) were significantly higher in the classical IP injection group than in the nasal drip groups. CONCLUSION: The IP injection model and the nasal-only delivery model are 2 typical models for AR that causes a different immune response. A positive dose-response relationship in the nasal-only delivery model is observed from 25 mg/mL to 0.025 mg/mL.


Assuntos
Alérgenos/administração & dosagem , Ovalbumina/administração & dosagem , Rinite Alérgica/imunologia , Administração Intranasal , Alérgenos/imunologia , Animais , Citocinas/imunologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Esquema de Medicação , Eosinófilos/patologia , Feminino , Imunoglobulina E/sangue , Injeções Intraperitoneais , Camundongos Endogâmicos BALB C , Líquido da Lavagem Nasal/imunologia , Mucosa Nasal/imunologia , Mucosa Nasal/patologia , Ovalbumina/imunologia , Rinite Alérgica/sangue
18.
Int Immunopharmacol ; 70: 260-267, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30851706

RESUMO

Nonylphenol (NP) is a widely distributed, toxic endocrine-disrupting chemical exhibiting estrogenic activity. However, its effect on allergic rhinitis (AR) remains unclear. In this study, the effects of NP on a murine model of AR were investigated. Mice were divided into ovalbumin (OVA), NP, and control groups. OVA was used for sensitization and challenge. Mice in the NP group were administered NP during the sensitization period. Allergic nasal symptoms and eosinophil counts in nasal mucosa were measured. Serum levels of OVA-specific IgE were determined by enzyme-linked immunosorbent assay. The mRNA levels of transcription factors of Th cells were determined with real-time polymerase chain reaction. Th cell subtypes and Treg numbers were counted with the aid of multi-color flow cytometry. Cytokine concentrations in nasal mucosa were determined using the cytometric bead array method. Subcutaneous injection of NP into mice exhibiting AR enhanced not only the nasal allergic symptoms, but also eosinophil infiltration and OVA-specific IgE. Moreover, NP upregulated IL-4, IL-5, IL-13, IL-9, IL-6 and IL-17, and downregulated IL-10, in the AR mouse model; IFN-γ and IL-23 were not affected. Transcription factors and Th cell percentages were evaluated to determine whether NP regulates Th cell subtypes in an AR mouse model. GATA3, PU.1, and RORγt levels were significantly increased, but FoxP3 and Helios were decreased. In addition, Th2, Th9, and Th17 subtype percentages significantly increased, and Treg cell percentages decreased, in NP administration groups; the percentage of Th1 subtypes was not affected. NP enhanced allergic inflammation in the AR mouse model through upregulation of Th2, Th9, and Th17 responses and negative regulation of Treg responses. These results suggest that NP may be trigger AR.


Assuntos
Poluentes Atmosféricos/efeitos adversos , Mucosa Nasal/imunologia , Fenóis/administração & dosagem , Rinite Alérgica/imunologia , Linfócitos T Reguladores/imunologia , Células Th17/imunologia , Células Th2/imunologia , Alérgenos/imunologia , Animais , Citocinas/metabolismo , Modelos Animais de Doenças , Fator de Transcrição GATA3/genética , Fator de Transcrição GATA3/metabolismo , Regulação da Expressão Gênica , Humanos , Imunomodulação , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Fenóis/efeitos adversos
19.
Int Immunopharmacol ; 70: 512-519, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30884431

RESUMO

Allergic rhinitis (AR) is an allergic nasal disease characterized by nasal obstruction, rhinorrhea, sneezing, and itching. Type 1 helper T cells (Th1)/type 2 helper T cells (Th2) imbalance has been identified as an important immunological mechanism of AR. In addition, up-regulation of type 17 helper T cells (Th17) also increase the risk of developing AR. Gallic acid (3, 4, 5-trihydroxybenzoic acid, GA), a polyphenol natural product, is obtained from various herbs, red wine, and green tea. It is known to have diverse biological effects such as anti-oxidation, anti-inflammation, anti-microbial and anti-cancer. In the present study, the effect of GA on airway inflammation and expression of Th1, Th2 and Th17 cytokines in an ovalbumin (OVA)-induced AR mouse model were investigated. GA alleviated the nasal allergic symptoms, reduced the thickness of nasal mucosa, attenuated goblet cell hyperplasia and eosinophil cell infiltration in the nasal mucosa, decreased the levels of interleukin (IL)-4, IL-5, IL-13 and IL-17 in nasal lavage fluid (NALF), and diminished the levels of OVA-specific IgE, OVA-specific IgG1 and OVA-specific IgG2a in serum. However, GA increased the expression of interferon-gamma and IL-12 in NALF. Taken together, it suggests that GA may be used as a therapeutic agent for AR.


Assuntos
Anti-Inflamatórios/uso terapêutico , Eosinófilos/imunologia , Ácido Gálico/uso terapêutico , Inflamação/tratamento farmacológico , Mucosa Nasal/imunologia , Rinite Alérgica/tratamento farmacológico , Células Th1/imunologia , Alérgenos/imunologia , Animais , Citocinas/metabolismo , Modelos Animais de Doenças , Humanos , Imunoglobulina E/metabolismo , Ativação Linfocitária , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Células Th17/imunologia , Células Th2/imunologia
20.
Int J Pharm ; 561: 187-196, 2019 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-30836154

RESUMO

Archaeosomes are liposomes composed of natural or synthetic archaeal lipids that can be used as adjuvants to induce strong long-lasting humoral and cell-mediated immune responses against entrapped antigen. However, the entrapment efficiency of antigen within archaeosomes constituted using standard liposome forming methodology is often only 5-40%. In this study, we evaluated different formulation methods using a simple semi-synthetic archaeal lipid (SLA, sulfated lactosyl archaeol) and two different antigens, ovalbumin (OVA) and hepatitis B surface antigen (HBsAg). Antigen was entrapped within archaeosomes using the conventional thin film hydration-rehydration method with or without removal of non-entrapped antigen, or pre-formed empty archaeosomes were simply admixed with an antigen solution. Physicochemical characteristics were determined (size distribution, zeta potential, vesicle morphology and lamellarity), as well as location of antigen relative to bilayer using cryogenic transmission electron microscopy (TEM). We demonstrate that antigen (OVA or HBsAg) formulated with SLA lipid adjuvants using all the different methodologies resulted in a strong antigen-specific immune response. Nevertheless, the advantage of using a drug substance process that comprises of simply admixing antigen with pre-formed empty archaeosomes, represents a simple, efficient and antigenic dose-sparing formulation for adjuvanting and delivering vaccine antigens.


Assuntos
Adjuvantes Imunológicos/química , Antígenos Arqueais/imunologia , Archaea/imunologia , Portadores de Fármacos/química , Lipídeos/química , Lipossomos/química , Vacinas/imunologia , Adjuvantes Imunológicos/farmacologia , Animais , Anticorpos/sangue , Contagem de Células , Fenômenos Químicos , Feminino , Antígenos de Superfície da Hepatite B/imunologia , Imunidade Celular/efeitos dos fármacos , Interferon gama/metabolismo , Lipossomos/ultraestrutura , Camundongos , Ovalbumina/imunologia , Baço/metabolismo , Vacinas/química
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