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2.
PLoS One ; 15(2): e0229121, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32101539

RESUMO

Since dogs play a central role in the contamination of humans and livestock with Echinococcus granulosus, the development of an effective vaccine for dogs is essential to control the disease caused by this parasite. For this purpose, a formulation based on biodegradable polymeric nanoparticles (NPs) as delivery system of recombinant Echinococcus granulosus antigen (tropomyosin EgTrp) adjuved with monophosphoryl lipid A (MPLA) has been developed. The obtained nanoparticles had a size of approximately 200 nm in diameter into which the antigen was correctly preserved and encapsulated. The efficiency of this system to deliver the antigen was evaluated in vitro on canine monocyte-derived dendritic cells (cMoDCs) generated from peripheral blood monocytes. After 48 h of contact between the formulations and cMoDCs, we observed no toxic effect on the cells but a strong internalization of the NPs, probably through different pathways depending on the presence or not of MPLA. An evaluation of cMoDCs activation by flow cytometry showed a stronger expression of CD80, CD86, CD40 and MHCII by cells treated with any of the tested formulations or with LPS (positive control) in comparison to cells treated with PBS (negative control). A higher activation was observed for cells challenged with EgTrp-NPs-MPLA compared to EgTrp alone. Formulations with MPLA, even at low ratio of MPLA, give better results than formulations without MPLA, proving the importance of the adjuvant in the nanoparticles structure. Moreover, autologous T CD4+ cell proliferation observed in presence of cMoDCs challenged with EgTrp-NPs-MPLA was higher than those observed after challenged with EgTrp alone (p<0.05). These first results suggest that our formulation could be used as an antigen delivery system to targeting canine dendritic cells in the course of Echinococcus granulosus vaccine development.


Assuntos
Antígenos de Protozoários/administração & dosagem , Células Dendríticas/imunologia , Cães/parasitologia , Equinococose/prevenção & controle , Echinococcus granulosus/imunologia , Vacinas Protozoárias/administração & dosagem , Adjuvantes Imunológicos/administração & dosagem , Animais , Antígenos de Protozoários/genética , Antígenos de Protozoários/imunologia , Linfócitos T CD4-Positivos/imunologia , Diferenciação Celular , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Células Dendríticas/efeitos dos fármacos , Cães/sangue , Cães/imunologia , Portadores de Fármacos/química , Portadores de Fármacos/toxicidade , Equinococose/imunologia , Equinococose/parasitologia , Equinococose/veterinária , Echinococcus granulosus/genética , Imunogenicidade da Vacina , Lipídeo A/análogos & derivados , Lipídeo A/química , Lipídeo A/toxicidade , Ativação Linfocitária/imunologia , Monócitos/fisiologia , Nanopartículas/química , Nanopartículas/toxicidade , Poliésteres/química , Poliésteres/toxicidade , Cultura Primária de Células , Vacinas Protozoárias/genética , Vacinas Protozoárias/imunologia , Testes de Toxicidade Aguda , Tropomiosina/administração & dosagem , Tropomiosina/genética , Tropomiosina/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia
3.
Toxicol Lett ; 322: 50-57, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-31958493

RESUMO

Allergic contact dermatitis (ACD) is an important occupational and environmental disease caused by topical exposure to chemical allergens. In the EU, it has been calculated that 4 % of animals are used in toxicity test for the assessment of skin sensitization (Peiser et al., 2012). To come a complete replacement of animals, evaluation of relative skin sensitization potency is necessary. The identification of mechanisms influencing allergen potency requires a better understanding of molecular events that trigger cell activation. Therefore, (i) the effects of selected allergens on surface markers expression and cytokines release in contact allergen-induced cell activation were assessed, and (ii) the role of Protein Kinase C (PKC) beta activation in contact allergen-induced cell activation was investigated. The human pro-myelocytic cell line THP-1 was used as experimental model surrogate of dendritic cells. Cells were exposed to select contact allergens of different potency and cell surface marker expression (CD80, CD86, HLA-DR) was determined by flow cytometry analysis. Cytokines production (IL-6, IL-8, IL-10, IL-12p40, IL-18) was evaluated with specific sandwich ELISA. The effective contribution of PKC beta in chemical allergen-induced cell activation was assessed by Western Blot analysis (PKC beta activation) and using a specific PKC beta inhibitor (PKC beta pseudosubstrate). In addition, to investigate if contact allergens are able to induce indeed dendritic cells (DCs) maturation, THP-1 cells were differentiated to immature DC and then exposed to contact allergen of different potency. Overall, our finding provides insights into the process of sensitization and strength of cell activation associated with allergens of different potency. Results obtained suggest that contact allergens of different potency are able to induce a different degree of activation of dendritic cells maturation involved in the process of ACD.


Assuntos
Alérgenos/classificação , Alternativas aos Testes com Animais , Células Dendríticas/efeitos dos fármacos , Dermatite Alérgica de Contato , Pele/efeitos dos fármacos , Xenobióticos/classificação , Alérgenos/toxicidade , Antígenos de Superfície/biossíntese , Biomarcadores/metabolismo , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Células Dendríticas/enzimologia , Células Dendríticas/imunologia , Dermatite Alérgica de Contato/enzimologia , Dermatite Alérgica de Contato/imunologia , Relação Dose-Resposta a Droga , Ativação Enzimática/efeitos dos fármacos , Humanos , Proteína Quinase C beta/metabolismo , Pele/enzimologia , Pele/imunologia , Xenobióticos/toxicidade
5.
Carbohydr Polym ; 229: 115457, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31826423

RESUMO

We previously demonstrated that porphyran, a sulfated polysaccharide extracted from Pyropia yezoensis, shows protective effects on LPS-induced septic shock in the mouse. However, the immune cell-mediated inhibitory effect of porphyran in LPS-induced activation of immune cells has not been well investigated. In this study, we found that treatment of porphyran suppressed LPS-induced upregulation of costimulatory molecule and C-C chemokine receptor type 7 (CCR7) expression in bone marrow-derived dendritic cells (BMDCs) in vitro and spleen DCs in vivo. Moreover, the LPS-induced expression of IL-6, IL-12, and TNF-α in the culture medium of BMDCs and serum dose-dependently decreased by porphyran treatment, which contributed to the inhibition of the intracellular cytokine production in spleen DCs. In addition, LPS-induced differentiation of helper T1 (Th1) and cytotoxic T1 (Tc1) cells was effectively suppressed by porphyran treatment in mice. The inhibitory effect of porphyran in LPS-induced immune activation was mediated by competitive binding of porphyran with LPS in spleen DCs. Thus, these results suggest that porphyran is a promising potential therapeutic agent in endotoxin-mediated inflammatory disease and septic shock.


Assuntos
Anti-Inflamatórios/farmacologia , Células Dendríticas/efeitos dos fármacos , Polissacarídeos Bacterianos/farmacologia , Sefarose/análogos & derivados , Animais , Células da Medula Óssea/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Citocinas/metabolismo , Imunidade Celular/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Camundongos Endogâmicos C57BL , Rodófitas/química , Sefarose/farmacologia , Baço/citologia , Linfócitos T Citotóxicos/metabolismo , Células Th1/metabolismo
6.
Immunology ; 159(1): 109-120, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31606893

RESUMO

Serpins are evolutionarily conserved serine protease inhibitors that are widely distributed in animals, plants and microbes. In this study, we reported the cloning and functional characterizations of two novel serpin genes, HlSerpin-a and HlSerpin-b, from the hard tick Haemaphysalis longicornis of China. Recombinant HlSerpin-a and HlSerpin-b displayed protease inhibitory activities against multiple mammalian proteases. Similar to other tick serpins, HlSerpin-a and HlSerpin-b suppressed the expression of inflammatory cytokines such as TNF-α, interleukin (IL)-6 and IL-1ß from lipopolysaccharide-stimulated mouse bone-marrow-derived macrophages (BMDMs) or mouse bone-marrow-derived dendritic cells (BMDCs). The minimum active region (reaction centre loop) of HlSerpin-a, named SA-RCL, showed similar biological activities as HlSerpin-a in the protease inhibition and immune suppression assays. The immunosuppressive activities of full-length HlSerpin-a and SA-RCL are impaired in Cathepsin G or Cathepsin B knockout mouse macrophages, suggesting that the immunomodulation functions of SA and SA-RCL are dependent on their protease inhibitory activity. Finally, we showed that both full-length HlSerpins and SA-RCL can relieve the joint swelling and inflammatory response in collagen-induced mouse arthritis models. These results suggested that HlSerpin-a and HlSerpin-b are two functional arthropod serpins, and the minimal reactive peptide SA-RCL is a potential candidate for drug development against inflammatory diseases.


Assuntos
Artrite Experimental/prevenção & controle , Proteínas de Artrópodes/farmacologia , Células Dendríticas/efeitos dos fármacos , Imunossupressores/farmacologia , Ixodidae/metabolismo , Articulações/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Serpinas/farmacologia , Animais , Artrite Experimental/imunologia , Artrite Experimental/metabolismo , Artrite Experimental/patologia , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/isolamento & purificação , Citocinas/metabolismo , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Imunossupressores/isolamento & purificação , Ixodidae/genética , Articulações/imunologia , Articulações/metabolismo , Articulações/patologia , Macrófagos/imunologia , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos DBA , Conformação Proteica , Células RAW 264.7 , Saliva/metabolismo , Serpinas/genética , Serpinas/isolamento & purificação , Relação Estrutura-Atividade
7.
APMIS ; 128(1): 10-19, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31642122

RESUMO

Atherogenesis is associated with chronic gut infections; however, the mechanisms are not clear. The aim of the study was to determine whether lipopolysaccharide of E. coli (E. coli LPS) may affect endothelial barrier and modify IL-10 expression in dendritic cells (DCs). Human umbilical vein endothelial cells (HUVECs) and monocyte-derived DCs were treated with E. coli LPS, apolipoprotein B100 (ApoB100) and 7-ketocholesterol (7-kCH) - harmful oxidized form of cholesterol. The effect of E. coli LPS, 7-kCH and ApoB100 on the barrier functions of HUVECs in real-time cell electric impedance sensing system (RTCA-DP) was assessed. Furthermore, the effect of 7-kCH and ApoB100 on barrier functions of HUVECs co-cultured with DCs previously treated with LPS was analyzed. Both E. coli LPS and 7-kCH decreased barrier functions of HUVECs and reduced tight junction protein mRNA expression, whereas ApoB100 increased endothelial barrier. In DCs, ApoB100 and E. coli LPS decreased IL-10 mRNA expression. In HUVECs co-cultured with DCs treated with LPS and subsequently pulsed with ApoB100 or 7-kCH, IL-10 mRNA expression was lower. E. coli LPS-exposed DCs diminished the protective effect of ApoB100 on endothelial integrity and led to the decrease in occludin mRNA expression. LPS potentially derived from gut microflora may destabilize endothelial barrier together with oxidized cholesterol and intensify the immunogenicity of ApoB100.


Assuntos
Células Dendríticas/efeitos dos fármacos , Escherichia coli/imunologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Interleucina-10/genética , Lipopolissacarídeos/imunologia , Junções Íntimas/efeitos dos fármacos , Apolipoproteína B-100/farmacologia , Células Cultivadas , Técnicas de Cocultura , Células Dendríticas/imunologia , Escherichia coli/química , Humanos , Cetocolesteróis/farmacologia , Lipopolissacarídeos/farmacologia , Ocludina/genética , Transdução de Sinais/efeitos dos fármacos , Proteínas de Junções Íntimas/genética
8.
Carbohydr Polym ; 229: 115403, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31826481

RESUMO

We examined the efficacy of fucoidan-N-(2-hydroxy-3-trimethylammonium)propylchitosan nanoparticles (FUC-HTCC NPs) as adjuvants for anthrax vaccine adsorbed (AVA). Positively and negatively surface-charged FUC-HTCC NPs were prepared via polyelectrolyte complexation by varying the mass ratio of FUC and HTCC. When cultured with L929 cells or JAWS II dendritic cells, both charged NPs showed high cell viability and low cytotoxicity, observed via MTT assay and lactate dehydrogenase release assay, respectively. In addition, we have monitored excellent NPs uptake efficacy by dendritic cells and observed that combining FUC-HTCC NPs with AVA significantly increases the magnitude of IgG-anti-protective antigen titers in A/J mice compared to that by CpG oligodeoxynucleotides plus AVA or AVA alone, and PA-specific IgG1 and IgG2a analysis confirmed that FUC-HTCC NPs strongly stimulated humoral immunity. Furthermore, FUC-HTCC NPs plus AVA provided a superior survival rate (100%) of A/J mice compared to CpG oligodeoxynucleotides plus AVA (75%) or AVA alone (50%) following anthrax lethal toxin challenge. The findings support FUC-HTCC NPs as a potential adjuvant of AVA for rapid induction of protective immunity.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Vacinas contra Antraz/administração & dosagem , Quitosana/administração & dosagem , Nanopartículas/administração & dosagem , Polissacarídeos/administração & dosagem , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/metabolismo , Feminino , Camundongos , Oligodesoxirribonucleotídeos
9.
Int J Nanomedicine ; 14: 8235-8249, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31802864

RESUMO

Background: The effective induction of an antigen-specific T cell immune response through dendritic cell activation is one of the key goals of tumor immunotherapy. Methods: In this study, efficient antigen-delivery carriers using silica-coated magnetic nanoparticles were designed and, their antigen-specific T cell immune response through dendritic cell activation investigated. Results: The results showed that the silica-coated magnetic nanoparticles with conjugated ovalbumin enhanced the production of cytokines and antigen uptake in bone marrow-derived dendritic cells. Also, this induced an antigen-specific cytotoxic T lymphocyte (CTL) immune response and activated antigen-specific Th1 cell responses, including IL-2 and IFN-γ production and proliferation. We proved that the immune-stimulatory effects of silica-coated magnetic nanoparticles with conjugated ovalbumin were efficient in inhibiting of tumor growth in EG7-OVA (mouse lymphoma-expressing ovalbumin tumor-bearing mice model). Conclusion: Therefore, the silica-coated magnetic nanoparticles with conjugated ovalbumin are expected to be useful as efficient anti-cancer immunotherapy agents.


Assuntos
Imunoterapia , Nanopartículas de Magnetita/química , Neoplasias/imunologia , Neoplasias/terapia , Ovalbumina/química , Dióxido de Silício/química , Animais , Antígenos/administração & dosagem , Galinhas , Citocinas/biossíntese , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Feminino , Imunidade , Nanopartículas de Magnetita/toxicidade , Camundongos Endogâmicos C57BL
10.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(11): 979-985, 2019 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-31878993

RESUMO

Objective To investigate the role of mitogen activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway in the salidroside (SAL)-regulated antitumor immunity of dendritic cells (DCs). Methods Lewis lung cancer-bearing mouse model was established and treated with normal saline (NS) (0.2 mL/d), SAL [500 mg/(kg.d)] and cyclophosphamide (CTX) [10 mg/(kg.d)] in corresponding groups. Then their survival and tumor quality were recorded. Meanwhile, the bone marrow-derived DCs from the Lewis lung cancer-bearing mice were isolated in vitro and cultured for 7 days before collection. After sorted with magnetic beads, cells were co-cultured with PBS, SAL (0.05 mg/mL), LPS (200 ng/mL) or SAL combined with ERK inhibitor U0126 separately for 48 hours, followed by detection of the expression of phosphorylated ERK (p-ERK) by flow cytometry. In addition, after the stimulation of DCs by SAL, the expression of p38 MAPK and phosphorylated c-jun N-terminal kinase (p-JNK) was detected by flow cytometry. Moreover, after sorted with magneticbeads, DCs were co-cultured with PBS, SAL (0.05 mg/mL), LPS (200 ng/mL) or SAL combined with U0126. Then, the supernatant was collected, and IL-12p70 secretion ability of DCs was detected by ELISA. Finally, sensitized bone marrow-derived DCs from Lewis lung cancer-bearing mice were stimulated with SAL and co-cultured with spleen T lymphocytes purified by nylon fiber column from C57BL/6 mice as effector cells. 3LL target cells were then added at the target ratio of 10:1, 25:1, 50:1, followed by the detection of cell viability of cytotoxic T lymphocytes (CTLs) using CCK-8 assay. Results SAL could significantly inhibit tumor growth and improve the survival rate. Compared with PBS group, the phosphorylation of ERK protein on DC was enhanced significantly, and then reduced significantly after U0126 treatment. The phosphorylation of p38MAPK and c-jun N-terminal kinase (JNK) protein was not statistically affected by SAL. The level of IL-12p70 significantly increased in SAL group, and became remarkably higher after U0126 treatment. Finally, the killing ability of CTL was significantly enhanced by SAL. Conclusion SAL can inhibit the tumor growth of Lewis lung cancer-bearing mice and activate the ERK signaling pathway, thereby promoting IL-12p70 secretion in DCs and enhancing the cytotoxicity of CTL.


Assuntos
Carcinoma Pulmonar de Lewis/imunologia , Células Dendríticas/efeitos dos fármacos , Glucosídeos/farmacologia , Sistema de Sinalização das MAP Quinases , Fenóis/farmacologia , Animais , Células Cultivadas , Células Dendríticas/imunologia , Interleucina-12/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Linfócitos T Citotóxicos/imunologia
11.
Pathologe ; 40(Suppl 3): 259-264, 2019 Dec.
Artigo em Alemão | MEDLINE | ID: mdl-31720747

RESUMO

Hyperosmolar micromilieu has been observed in physiologic (kidney medulla, lymphatic tissue) and pathologic (renal allorejection, solid tumors) conditions. Hyperosmolarity can modulate gene expression and alter the stimulatory profile of macrophages and dendritic cells. We have reported that dendritic cells upon exposure to hypertonic stimuli shift their profile towards a macrophage-M2-like phenotype, resulting in attenuated local alloreactivity during acute kidney graft rejection. Moreover, we showed that a hyperosmotic microenvironment affects the cross-priming capacity of dendritic cells. Using ovalbumin as a model antigen, we showed that exposure of dendritic cells to hyperosmolarity strongly inhibits activation of antigen-specific T cells despite enhancement of antigen uptake, processing, and presentation; it can reduce dendritic cell-T cell contact time. We have identified TRIF as key mediator of this phenomenon. Moreover, we detected a hyperosmolarity-triggered, TRIF-dependent clustering of MHC class I­antigen complexes, but not of unloaded MHCI molecules, providing a possible explanation for a reduced T cell activation. Our findings identify dendritic cells as important players in hyperosmolarity-triggered immune imbalance and suggest that targeting local hyperosmolarity in tumor micromilieu may contribute to an enhanced specific anti-tumor immune response.


Assuntos
Proteínas Adaptadoras de Transporte Vesicular/metabolismo , Células Dendríticas , Antígenos de Histocompatibilidade Classe I , Apresentação Cruzada , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Antígenos de Histocompatibilidade Classe I/metabolismo , Humanos , Ativação Linfocitária , Osmorregulação , Ovalbumina/imunologia , Linfócitos T
12.
J Agric Food Chem ; 67(48): 13333-13343, 2019 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-31703480

RESUMO

Fluoride (F) widely exists in the water and food. Recent studies reported that F induced testicular toxicity via inflammation reaction. This study was aimed to explore the mechanism of F-induced inflammation in testis. 100 healthy male mice (BALB/cJ strain) were randomly divided into five groups including: control, experimental autoimmune orchitis (EAO), and three F groups (25, 50, and 100 mg/L sodium fluoride (NaF)). After 150 d, the results showed a significant increase in testicular cytokines levels including of IL-17A, IL-6, IFN-γ, and TNF-α in NaF and EAO groups compared with control group. Interestingly, the presence of specific antisperm autoantibodies in antitesticular autoantibodies and the notable recruitment of immunocyte (T cells and dendritic cells) were also observed in NaF and EAO groups. In addition, findings showed that in NaF and EAO groups macrophages and T cells both significantly secreted IL-17A, and the protein and mRNA levels of cytokines (IL-6 and TGF-ß) were significantly increased. From these results, it can be concluded that autoimmune orchitis and IL-17A are implicated in F-induced testicular inflammation.


Assuntos
Doenças Autoimunes/induzido quimicamente , Doenças Autoimunes/imunologia , Fluoretos/efeitos adversos , Interleucina-17/imunologia , Orquite/imunologia , Testículo/imunologia , Animais , Doenças Autoimunes/genética , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Humanos , Interleucina-17/genética , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Orquite/induzido quimicamente , Orquite/genética , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Testículo/efeitos dos fármacos , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia
13.
J Agric Food Chem ; 67(43): 12137-12143, 2019 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-31566976

RESUMO

This study evaluated the immunomodulatory effects of two high-molecular-weight and structurally different mushroom polysaccharides, an alkali-soluble polysaccharide (mPRSon) and a water-soluble polysaccharide-protein complex (PRW), isolated previously from the sclerotia of Pleurotus rhinocerus, on the maturation of murine bone-marrow-derived dendritic cells (BMDCs). The effects of mPRSon and PRW on the expression of morphological change, surface molecules, phagocytic activity, and cytokine release in BMDCs were determined by flow cytometry and a mouse cytokine array. The results showed that both mPRSon and PRW could induce phenotypic and functional maturation of BMDCs. At the same time, mPRSon upregulated the expression of membrane phenotypic marker CD86 and PRW markedly upregulated CD40, CD80, and CD86. In addition, mPRSon could bind to the dectin-1 receptor and stimulate the release of MIP-1α, MIP-2, and IL-2, while PRW could bind to complement receptor 3 and toll-like receptor 2 with an upregulation of the expression of IL-2, IL-6, MIP-1α, MIP-2, RANTES, IL-12p40p70, IL-12p70, TIMP-1, IFN-γ, KC, MCP-1, and GCSF. The study provides additional information on how structural differences in sclerotial polysaccharides influence their immunomodulatory activities on BMDCs involving different PAMP receptors. It is anticipated that more understanding of the interactions between the sclerotial polysaccharides and their receptors in immune cells can facilitate their future application for cancer immunotherapy.


Assuntos
Células da Medula Óssea/efeitos dos fármacos , Células Dendríticas/efeitos dos fármacos , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Polyporus/química , Polissacarídeos/química , Polissacarídeos/farmacologia , Animais , Células da Medula Óssea/imunologia , Células Cultivadas , Citocinas/genética , Citocinas/imunologia , Células Dendríticas/imunologia , Fatores Imunológicos/isolamento & purificação , Camundongos , Camundongos Endogâmicos BALB C , Extratos Vegetais/isolamento & purificação , Polissacarídeos/isolamento & purificação
14.
Int J Nanomedicine ; 14: 7053-7064, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31564865

RESUMO

Background: Food allergy (FA) is a significant public health problem. The therapeutic efficacy for FA is unsatisfactory currently. The breakdown of intestinal immune tolerance is associated with the pathogenesis of FA. Therefore, it is of great significance to develop novel therapeutic methods to restore immune tolerance in treating FA. Methods: We proposed an oral administration strategy to treat FA by co-delivering food allergen epitope fragment (peptide: IK) and adjuvant R848 (TLR7 ligand) in the mPEG-PDLLA nanoparticles (PPLA-IK/R848 NPs). The generation of tolerogenic dendritic cells (DCs) and regulatory T cells (Tregs) induced by PPLA-IK/R848 NPs were evaluated in vitro and in vivo. The therapeutic effects of PPLA-IK/R848 NPs were also assessed in an OVA-induced FA model. Results: PPLA-IK/R848 NPs could efficiently deliver IK to DCs to drive DCs into the tolerogenic phenotypes and promote the differentiation of Tregs in vitro and in vivo, significantly inhibited FA responses through the recovery of intestinal immune tolerance. Conclusion: Oral administration of PPLA-IK/R848 NPs could efficiently deliver IK and R848 to intestinal DCs and stimulate DCs into allergen tolerogenic phenotype. These tolerogenic DCs could promote the differentiation of Tregs, which significantly protected mice from food allergic responses. This study provided an efficient formulation to alleviate FA through the recovery of immune tolerance.


Assuntos
Alérgenos/imunologia , Células Dendríticas/imunologia , Sistemas de Liberação de Medicamentos , Epitopos/imunologia , Hipersensibilidade Alimentar/tratamento farmacológico , Hipersensibilidade Alimentar/imunologia , Imidazóis/administração & dosagem , Imidazóis/uso terapêutico , Tolerância Imunológica , Linfócitos T Reguladores/imunologia , Sequência de Aminoácidos , Animais , Diferenciação Celular , Células Dendríticas/efeitos dos fármacos , Imidazóis/química , Imidazóis/farmacologia , Tolerância Imunológica/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Peptídeos/química , Poliésteres/química , Polietilenoglicóis/química , Linfócitos T Reguladores/efeitos dos fármacos
15.
Int J Mol Sci ; 20(18)2019 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-31533251

RESUMO

Dendritic cells (DCs) and leukemia-derived DC (DCleu) are potent stimulators of various immunoreactive cells and they play a pivotal role in the (re-) activation of the immune system. As a potential treatment tool for patients with acute myeloid leukemia, we developed and analyzed two new PGE1-containing protocols (Pici-PGE1, Kit M) to generate DC/DCleu ex vivo from leukemic peripheral blood mononuclear cells (PBMCs) or directly from leukemic whole blood (WB) to simulate physiological conditions. Pici-PGE1 generated significantly higher amounts of DCs from leukemic and healthy PBMCs when compared to control and comparable amounts as the already established protocol Pici-PGE2. The proportions of sufficient DC-generation were even higher after DC/DCleu-generation with Pici-PGE1. With Kits, it was possible to generate DCs and DCleu directly from leukemic and healthy WB without induction of blast proliferation. The average amounts of generated DCs and DCleu-subgroups were comparable with all Kits. The PGE1 containing Kit M generated significantly higher amounts of mature DCs when compared to the PGE2-containing Kit K and increased the anti-leukemic-activity. In summary PGE1-containing protocols were suitable for generating DC/DCleu from PBMCs as well as from WB, which reliably (re-) activated immunoreactive cells, improved the overall ex vivo anti-leukemic activity, and influenced cytokine-release-profiles.


Assuntos
Alprostadil/farmacologia , Células Dendríticas/citologia , Células Dendríticas/efeitos dos fármacos , Adulto , Idoso , Biomarcadores , Diferenciação Celular/efeitos dos fármacos , Citocinas/metabolismo , Feminino , Citometria de Fluxo , Humanos , Imunomodulação/efeitos dos fármacos , Imunofenotipagem , Leucemia Mieloide Aguda/metabolismo , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Masculino , Pessoa de Meia-Idade , Picibanil/farmacologia , Adulto Jovem
16.
Food Chem Toxicol ; 133: 110793, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31473338

RESUMO

The toxicity of dietary E 171, a food grade titanium dioxide was evaluated. A recent study reported rats receiving E 171 in water developed inflammation and aberrant crypt foci (ACF) in the gastrointestinal tract. Here, rats received food containing E 171 (7 or 100 days). The 100-day study included feeding E 171 after dimethylhydrazine (DMH) or vehicle only pretreatment. Food consumption was similar between treatment groups with maximum total cumulative E 171 exposure being 2617 mg/kg in 7 days and 29,400 mg/kg in 100 days. No differences were observed due to E 171 in the percentage of dendritic, CD4+ T or Treg cells within Peyer's patches or the periphery, or in cytokine production in plasma, sections of jejunum, and colon in 7- or 100-day E 171 alone fed rats. Differences were observed for IL-17A in colon (400 ppm E 171 + DMH) and IL-12p70 in plasma (40 ppm E 171 + DMH). E 171 had no effect on histopathologic evaluations of small and large intestines, liver, spleen, lungs, or testes, and no effects on ACF, goblet cell numbers, or colonic gland length. Dietary E 171 administration (7- or 100-day), even at high doses, produced no effect on the immune parameters or tissue morphology.


Assuntos
Aditivos Alimentares/toxicidade , Mucosa Intestinal/efeitos dos fármacos , Titânio/toxicidade , 1,2-Dimetilidrazina/farmacologia , Animais , Linfócitos T CD4-Positivos/efeitos dos fármacos , Carcinogênese/efeitos dos fármacos , Carcinógenos/farmacologia , Citocinas/metabolismo , Células Dendríticas/efeitos dos fármacos , Aditivos Alimentares/química , Masculino , Tamanho da Partícula , Nódulos Linfáticos Agregados/efeitos dos fármacos , Ratos Wistar , Linfócitos T Reguladores/efeitos dos fármacos , Titânio/química
17.
Eur Cytokine Netw ; 30(2): 43-58, 2019 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-31486403

RESUMO

The present study demonstrates that monocyte-derived dendritic cells (moDCs) produced in vitro using a GM-CSF and IFN-α differentiation protocol encompass a rare (∼5%) subpopulation of cells showing classical dendritic cell morphology and capable of natural internalization of extracellular self-DNA. We established that DEFB, HMGB1, LL-37 and RAGE antigens, which mediate the process of DNA internalization, are expressed on the surface of moDCs similar to plasmacytoid dendritic cells. However, in constrast to the latter subpopulation, these cells do not produce interleukin (IL)-37. Nonetheless, the process of DNA internalization was not in direct relation to the presence of the above antigens on the surface of these cells. Dendritic cells were sorted into total and non-DNA-internalizing populations and cytokine production was analyzed at 24-48 hours post-DNA treatment. We show that massive secretion of cytokines by dendritic cells is associated with the dsDNA-internalizing subpopulation. A total pool of IFN-moDCs secrete pro-inflammatory "first-wave" cytokines (IL-2, IL-6, IL-8, TNF-α) at both 24 and 48 hours time points. The anti-inflammatory cytokines IL-4 and IL-10 were found to be modestly induced, whereas GM-CSF, G-CSF, and IFN-γ production was strongly induced. Treatment of moDCs with dsDNA results in the up-regulated transcription of IFN-α, IFN-ß, IFN-γ, IL-8, IL-10, and VEGF by 6 hours. Combined dsDNA + chloroquine treatment has a synergistic effect on transcription of only one of the genes tested, with the pro-inflammatory cytokine IFN-ß displaying the strongest fold induction by 24 hours.


Assuntos
DNA/metabolismo , Células Dendríticas/citologia , Endocitose , Espaço Extracelular/metabolismo , Monócitos/citologia , Antígenos de Neoplasias/metabolismo , Peptídeos Catiônicos Antimicrobianos/metabolismo , Cloroquina/farmacologia , Citocinas/metabolismo , Sondas de DNA/metabolismo , Células Dendríticas/efeitos dos fármacos , Endocitose/efeitos dos fármacos , Feminino , Proteína HMGB1/metabolismo , Humanos , Interferons/farmacologia , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Monócitos/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Rodaminas/metabolismo , beta-Defensinas/metabolismo
18.
Cancer Immunol Immunother ; 68(10): 1605-1619, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31531696

RESUMO

The main effectors in tumor control are the class I MHC molecule-restricted CD8+ cytotoxic T lymphocytes (CTLs). Tumor-specific CTL induction can be regulated by dendritic cells (DCs) expressing both tumor-derived epitopes and co-stimulatory molecules. Immunosuppressive tolerogenic DCs, having down-regulated co-stimulatory molecules, are seen within the tumor mass and can suppress tumor-specific CTL induction. The tolerogenic DCs expressing down-regulated XCR1+CD141+ appear to be induced by tumor-derived soluble factors or dexamethasone, while the immunogenic DCs usually express XCR1+CD141+ molecules with a cross-presentation function in humans. Thus, if tolerogenic DCs can be reactivated into immunogenic DCs with sufficient co-stimulatory molecules, tumor-specific CD8+ CTLs can be primed and activated in vivo. In the present study, we converted human tolerogenic CD141+ DCs with enhanced co-stimulatory molecule expression of CD40, CD80, and CD86 through stimulation with non-toxic mycobacterial lipids such as mycolic acid (MA) and lipoarabinomannan (LAM), which synergistically enhanced both co-stimulatory molecule expression and interleukin (IL)-12 secretion by XCR1+CD141+ DCs. Moreover, MA and LAM-stimulated DCs captured tumor antigens and presented tumor epitope(s) in association with class I MHCs and sufficient upregulated co-stimulatory molecules to prime naïve CD3+ T cells to become CD8+ tumor-specific CTLs. Repeat CD141+ DC stimulation with MA and LAM augmented the secretion of IL-12. These findings provide us a new method for altering the tumor environment by converting tolerogenic DCs to immunogenic DCs with MA and LAM from Mycobacterium tuberculosis.


Assuntos
Células Dendríticas/imunologia , Lipopolissacarídeos/farmacologia , Mycobacterium/química , Ácidos Micólicos/farmacologia , Neoplasias/imunologia , Linfócitos T Citotóxicos/imunologia , Antígenos de Superfície/análise , Linhagem Celular Tumoral , Células Dendríticas/efeitos dos fármacos , Humanos , Interleucina-12/biossíntese , Mycobacterium bovis
19.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 35(7): 583-588, 2019 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-31537241

RESUMO

Objective To explore the effect of prostaglandin E2 (PGE2) on the phenotype and chemotaxis of mouse bone marrow-derived dendritic cells (BMDCs). Methods BMDCs isolated from murine bone marrow and cultured in vitro were divided into 6 groups (0 µg/mL PGE2 group, 1 µg/mL PGE2 group, 5 µg/mL PGE2 group, 0 µg/mL PGE2 plus LPS group, 1 µg/mL PGE2 plus LPS group, 5 µg/mL PGE2 plus LPS group). The expression of surface makers CD40, CD86, major histocompatibility complex II (MHCII), CC chemokine receptor 7 (CCR7) were detected by flow cytometry. The expression of CCR7 protein was detected by Western blot analysis. The migration ability of BMDCs was detected by TranswellTM assay. The survival rate of BMDCs was detected by CCK-8 assay. Results PGE2 of 1 µg/mL increased the expression of surface molecules on BMDCs and promoted the migration ability of BMDCs. PGE2 of 5 µg/mL reduced the expression of surface molecules on BMDCs and inhibited the migration ability of BMDCs. The change of PGE2 concentration did not affect the survival rate of BMDCs. Conclusion PGE2 was demonstrated a dual regulatory effect on the migration of BMDCs. Low concentration of PGE2 can promote the migration ability of BMDCs, while high concentration of PGE2 shows contrary effect.


Assuntos
Medula Óssea , Movimento Celular , Células Dendríticas/citologia , Dinoprostona/farmacologia , Animais , Antígeno B7-2/metabolismo , Antígenos CD40/metabolismo , Células Cultivadas , Células Dendríticas/efeitos dos fármacos , Antígenos de Histocompatibilidade Classe II/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Receptores CCR7/metabolismo
20.
Int J Mol Sci ; 20(15)2019 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-31382455

RESUMO

Recently, nanofibers (NFs) formed from antigenic peptides conjugated to ß-sheet-forming peptides have attracted much attention as a new generation of vaccines. However, studies describing how the hydrophilic-hydrophobic balance of NF components affects cellular interactions of NFs are limited. In this report, three different NFs were prepared by self-assembly of ß-sheet-forming peptides conjugated with model antigenic peptides (SIINFEKL) from ovalbumin and hydrophilic oligo-ethylene glycol (EG) of differing chain lengths (6-, 12- and 24-mer) to investigate the effect of EG length of antigen-loaded NFs on their cellular uptake, cytotoxicity, and dendritic cell (DC)-stimulation ability. We used an immortal DC line, termed JAWS II, derived from bone marrow-derived DCs of a C57BL/6 p53-knockout mouse. The uptake of NFs, consisting of the EG 12-mer by DCs, was the most effective and activated DC without exhibiting significant cytotoxicity. Increasing the EG chain length significantly reduced cellular entry and DC activation by NFs. Conversely, shortening the EG chain enhanced DC activation but increased toxicity and impaired water-dispersibility, resulting in low cellular uptake. These results show that the interaction of antigen-loaded NFs with cells can be tuned by the EG length, which provides useful design guidelines for the development of effective NF-based vaccines.


Assuntos
Adjuvantes Imunológicos/farmacologia , Antígenos/farmacologia , Células Dendríticas/efeitos dos fármacos , Ovalbumina/farmacologia , Peptídeos/farmacologia , Adjuvantes Imunológicos/química , Sequência de Aminoácidos , Animais , Antígenos/química , Linhagem Celular , Células Cultivadas , Células Dendríticas/imunologia , Etilenoglicol/química , Etilenoglicol/farmacologia , Interações Hidrofóbicas e Hidrofílicas , Camundongos Endogâmicos C57BL , Nanofibras/química , Nanofibras/ultraestrutura , Ovalbumina/química , Peptídeos/química , Conformação Proteica em Folha beta
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