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1.
Bull Exp Biol Med ; 166(6): 744-746, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31020588

RESUMO

We studied the effects of Ergoferon on the production of antiviral cytokine IL-2 by type 1 CD4+T cells. Preincubation of Jurkat cells with Ergoferon increased IL-2 secretion by these cells after stimulation with phorbol 12-myristate 13-acetate/ionomycine in comparison with the placebo group. The data prove that Ergoferon is capable of activating cell cascades involved in the realization of the antiviral immune response.


Assuntos
Anticorpos/farmacologia , Fatores Imunológicos/farmacologia , Interleucina-2/genética , Expressão Gênica , Humanos , Interleucina-2/biossíntese , Interleucina-2/imunologia , Ionomicina/farmacologia , Células Jurkat , Ativação Linfocitária/efeitos dos fármacos , Acetato de Tetradecanoilforbol/farmacologia
2.
Mater Sci Eng C Mater Biol Appl ; 98: 472-481, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30813049

RESUMO

Cyclosporine A (CsA) is an extremely hydrophobic immunosuppressive drug, whose systemic administration to suppress the activity of T cells and T cell-based immune responses is frequently associated with a number of adverse drug reactions. Local delivery of CsA focused on a specific target organ has been proposed as a possible solution to this problem. In this study, we developed biodegradable sol-gel drug delivery systems, consisting of HA-Ca-Alg hydrogels combining hyaluronic acid calcium complex (HA-Ca) and sodium alginate (Alg-Na) components, for the local sustained delivery of CsA. A HA-Ca complex with very high degree of substitution was prepared by the acid-base reaction of hyaluronic acid and calcium acetate. The gelation was completed within about 2-45 min without external addition of calcium salts such as CaSO4 and CaCl2, indicating the high potential of the present hydrogel systems for drug delivery by injection in vivo. The HA-Ca system was characterized by high-resolution inductively coupled plasma-optical emission spectroscopy, 1H NMR, FT-IR, and thermogravimetric analysis methods. Moreover, the scanning electron microscopy analysis of the HA-Ca-Alg hydrogels showed an irregular porous morphology, with interconnected pores of 50-300 µm width. The sol-gel transition and the maximum viscosity (about 10,000 cP) of the HA-Ca-Alg hydrogels were characterized by examining the time evolution of the viscosity at 37 °C. The hydrolytic degradation of the HA-Ca-Alg hydrogel was also examined at 37 °C. CsA-encapsulated HA-Ca-Alg hydrogels exhibited sustained in vitro release of CsA over 14 days, which was confirmed through in vitro measurements of the activity of murine T cells over 2 weeks. These results show that the present injectable HA-Ca-Alg hydrogels can be used effectively for the sustained delivery of extremely hydrophobic immunosuppressive drugs, including CsA.


Assuntos
Materiais Biocompatíveis/química , Sistemas de Liberação de Medicamentos , Hidrogéis/química , Imunossupressores/administração & dosagem , Injeções , Alginatos/química , Animais , Cálcio/química , Proliferação de Células/efeitos dos fármacos , Ciclosporina/administração & dosagem , Ciclosporina/farmacologia , Feminino , Ácido Hialurônico/química , Interleucina-2/biossíntese , Camundongos Endogâmicos C57BL , Espectroscopia de Prótons por Ressonância Magnética , Espectroscopia de Infravermelho com Transformada de Fourier , Linfócitos T/citologia , Linfócitos T/efeitos dos fármacos , Termogravimetria , Viscosidade
3.
Biochem Biophys Res Commun ; 509(4): 918-924, 2019 02 19.
Artigo em Inglês | MEDLINE | ID: mdl-30642628

RESUMO

Transient receptor potential (TRP) channels are a family of non-selective cation channels that are functionally expressed in various organs and cells. Among them, transient receptor potential vanilloid (TRPV) 1 and TRPV4 channels are expressed in T cells, where they serve as Ca2+ channels for T-cell receptor signaling [Bertin et al., 2014, Majhi et al., 2015]. Here, we show that not only TRPV1 and TRPV4 channel inhibitors, but also a transient receptor potential melastatin (TRPM) 8 channel inhibitor can suppress murine T-cell activation. Mouse splenic lymphocytes pretreated with N-(3-aminopropyl)-2-[(3-methylphenyl)methoxy]-N-(2-thienylmethyl)benzamide hydrochloride (AMTB), a TRPM8 channel-selective inhibitor, showed significantly reduced IL-2 and IL-6 release from T cells after stimulation with anti-CD3ε/anti-CD28 antibodies or concanavalin A. AMTB also suppressed IL-2 mRNA expression and activation of extracellular signal-regulated kinase 1/2, which is involved in IL-2 production. Further, the increase of CD25 (IL-2 receptor alpha chain) expression after T-cell activation was suppressed by AMTB. TRPM8 channel was expressed in CD4+ T cells isolated from splenocytes, and we confirmed that the release of IL-2 from isolated CD4+ T cells was significantly suppressed by AMTB. In vitro re-stimulation of splenocytes from external antigen-immunized mice with the same antigen induced IL-2 and IL-6 production, which was significantly suppressed by AMTB. Thus, the TRPM8 channel inhibitor AMTB suppresses T-cell activation induced by various stimulants.


Assuntos
Antígenos/metabolismo , Benzamidas/farmacologia , Concanavalina A/farmacologia , Ativação Linfocitária/efeitos dos fármacos , Receptores de Antígenos de Linfócitos T/metabolismo , Canais de Cátion TRPM/antagonistas & inibidores , Tiofenos/farmacologia , Animais , Linfócitos T CD4-Positivos/metabolismo , Interleucina-2/biossíntese , Interleucina-6/biossíntese , Camundongos , Baço/citologia
4.
J Leukoc Biol ; 105(1): 163-175, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30371945

RESUMO

IL-10 is a pleiotropic cytokine with immunoregulatory functions affecting various cell types. In a model of experimental infection with the protozoan Trypanosoma cruzi (T. cruzi), we found increased morbidity and lower parasite control in IL-10 deficient mice (IL-10 KO) compared to wild-type (WT) mice. Despite enhanced Mϕ function and dendritic cell activation, IL-10 KO mice were more susceptible to infection. The kinetics of T cells in spleen and peripheral blood revealed that infected IL-10 KO mice failed to increase the number of spleen and circulating total CD8+ T cells, a phenomenon observed from the second week of infection in WT mice. Total CD8+ T cells from IL-10 KO mice exhibited diminished proliferation, cytotoxic potential and IFN-γ production than their WT counterparts and T. cruzi-specific CD8+ T cells displayed reduced in vivo cytotoxicity. The absence of IL-10 selectively affected expansion, survival, and increased PD-1 expression of CD8+ T cells without altering these same parameters on CD4+ T cells. Increased inhibitory receptors expression and down-modulation of T-bet by CD8+ T cells from IL-10 KO infected mice were compatible with a T cell exhaustion phenotype. Collectively, these findings reveal that during acute infection, IL-10 plays a previously unrecognized stimulatory role on CD8+ T cells, the most relevant lymphocyte population for the control of intracellular T. cruzi stages. A clear knowledge of the underlying mechanisms that drive effector functions of cytotoxic T cells is critical to understand pathogen persistence and rational design of prophylactic strategies against T. cruzi.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Doença de Chagas/imunologia , Doença de Chagas/parasitologia , Interleucina-10/metabolismo , Ativação Linfocitária/imunologia , Trypanosoma cruzi/patogenicidade , Doença Aguda , Animais , Apoptose/efeitos dos fármacos , Linfócitos T CD8-Positivos/efeitos dos fármacos , Contagem de Células , Proliferação de Células/efeitos dos fármacos , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/metabolismo , Regulação para Baixo/efeitos dos fármacos , Interleucina-2/biossíntese , Ativação Linfocitária/efeitos dos fármacos , Subpopulações de Linfócitos/imunologia , Camundongos Endogâmicos BALB C , Camundongos Knockout , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Fenótipo , Receptor de Morte Celular Programada 1/metabolismo , Receptores de Interleucina-10/metabolismo , Proteínas Recombinantes/farmacologia , Baço/patologia , Virulência
5.
Nat Prod Res ; 33(14): 2024-2031, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29882435

RESUMO

Two new glycosides of phytolaccagenin and 2ß-hydroxyoleanoic acid, namely bonushenricoside A (3) and bonushenricoside B (5) together with four known saponins, respectively compounds 3-O-L-α-arabinopyranosyl-bayogenin-28-O-ß-glucopyranosyl ester (1), 3-O-ß-glucuronopyranosyl-2ß-hydroxygypsogenin-28-O-ß-glucopyranosyl ester (2), 3-O-ß-glucuronopyranosyl-bayogenin-28-O-ß-glucopyranosyl ester (4) and 3-O-ß-glucuronopyranosyl-medicagenic acid-28-ß-xylopyranosyl(1→4)-α-rhamnopyranosyl(1→2)-α-arabinopyranosyl ester (6) were isolated from the roots of Chenopodium bonus-henricus L. The structures of the compounds were determined by means of spectroscopic methods (1D and 2D NMR, IR and HRMS). The MeOH extract and compounds were tested for cytotoxic activity on five leukemic cell lines (HL-60, SKW-3, Jurkat E6-1, BV-173 and K-562). In addition, the ability of metanolic extract and saponins to modulate the interleukin-2 production in PHA/PMA stimulated Jurkat E6-1 cells was investigated as well.


Assuntos
Chenopodium/química , Saponinas/isolamento & purificação , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Glicosídeos/química , Humanos , Interleucina-2/biossíntese , Leucemia/tratamento farmacológico , Leucemia/patologia , Estrutura Molecular , Extratos Vegetais/química , Saponinas/química , Análise Espectral
6.
Bioorg Med Chem ; 27(2): 442-446, 2019 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-30579802

RESUMO

A new flavoalkaloid racemate, leucoflavonine (1), together with its flavonoid precursor pectolinarigenin (2), was isolated from the leaves of Leucosceptrum canum collected from Tibet. Its structure was established by comprehensive spectroscopic analysis. Chrial separation of the enantiomers of 1 was achieved, and their absolute configurations were determined as S-(+)- and R-(-)-leucoflavonines ((+)-1a and (-)-1b) by comparison of their computational and experimental optical rotations. Biological assays indicated that both (+)-1a and (-)-1b exhibited inhibitory activity against acetylchlorinesterase (AChE) in vitro (IC50 = 68.0 ±â€¯8.6 and 18.3 ±â€¯1.8 µM, respectively). Moreover, (-)-1b displayed cytotoxicity against human hepatoma cells HepG2 (IC50 = 52.9 ±â€¯3.6 µM), and inhibited the production of interleukelin-2 (IL-2) in Jurkat cells (IC50 = 16.5 ±â€¯0.9 µM), while (+)-1a showed no obvious activity in these assays.


Assuntos
Acetilcolinesterase/metabolismo , Alcaloides/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Inibidores da Colinesterase/farmacologia , Flavonas/farmacologia , Interleucina-2/biossíntese , Lamiaceae/química , Folhas de Planta/química , Alcaloides/química , Alcaloides/isolamento & purificação , Animais , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Proliferação de Células/efeitos dos fármacos , Inibidores da Colinesterase/química , Inibidores da Colinesterase/isolamento & purificação , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Enguias , Flavonas/química , Células Hep G2 , Humanos , Células Jurkat , Camundongos , Estrutura Molecular , Células RAW 264.7 , Relação Estrutura-Atividade
7.
Bull Exp Biol Med ; 166(2): 225-228, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30488211
8.
Sci Rep ; 8(1): 16123, 2018 10 31.
Artigo em Inglês | MEDLINE | ID: mdl-30382149

RESUMO

Interleukin-2 (IL-2) is a key regulator of adaptive immune responses but its regulation is incompletely understood. We previously found that PDL1-dependent signals were pivotal for liver sinusoidal endothelial cell-mediated priming of CD8 T cells, which have a strongly reduced capacity to produce IL-2. Here, we show that the expression of the ARF-like GTPase Arl4d is PD-L1-dependently induced in such LSEC-primed T cells, and is associated with reduced IL-2 secretion and Akt phosphorylation. Conversely, Arl4d-deficient T cells overproduced IL-2 upon stimulation. Arl4d-deficiency in CD8 T cells also enhanced their expansion and effector function during viral infection in vivo. Consistent with their increased IL-2 production, Arl4d-deficient T cells showed enhanced development into KLRG1+CD127- short-lived effector cells (SLEC), which is dependent on IL-2 availability. Thus, our data reveal a PD-L1-dependent regulatory circuitry that involves the induction of Arl4d for limiting IL-2 production in T cells.


Assuntos
Fatores de Ribosilação do ADP/metabolismo , Antígeno B7-H1/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Interleucina-2/biossíntese , Fatores de Ribosilação do ADP/deficiência , Adenoviridae/fisiologia , Animais , Linfócitos T CD8-Positivos/virologia , Diferenciação Celular , Proliferação de Células , Células Dendríticas/metabolismo , Células Endoteliais/metabolismo , Interleucina-2/metabolismo , Fígado/citologia , Ativação Linfocitária/imunologia , Camundongos Endogâmicos C57BL , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo
9.
In Vivo ; 32(6): 1353-1359, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30348688

RESUMO

BACKGROUND/AIM: Novel information on the role of endogenous compounds in regulating physiological and pathological process are of interest, as it may lead to the development of better strategies for disease management. The role of angiotensin II and the signaling of type 1 angiotensin II receptor (AGT1R) in T-lymphocyte activation and interleukin-2 (IL-2) production are largely unknown. MATERIALS AND METHODS: Jurkat T-cells were treated with AGT1R inhibitor candesartan and stimulated with phorbol myristate acetate (PMA) and ionomycin. T-Cell activation, associated cytokine production and levels of signaling proteins were evaluated by flow cytometry and western blot analysis. RESULTS: Candesartan significantly suppressed PMA and ionomycin-induced CD25 expression and IL-2 production. Regarding the molecular mechanism involved, we showed that such suppressive effects of blocking of AGT1R by candesartan resulted in the significant inhibition of ERK activation in PMA-stimulated Jurkat T-cells. The effect of ERK inhibition on T-cell activation was further confirmed. Treatment with FR180204, a specific ERK inhibitor, reduced T-cell activation and IL-2 secretion. CONCLUSION: AGT1R signaling is essential for T-cell activation and IL-2 production, and the inhibition of this pathway suppressed T-cell activation via an ERK-dependent mechanism.


Assuntos
Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Interleucina-2/biossíntese , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/imunologia , Receptor Tipo 1 de Angiotensina/metabolismo , Linfócitos T/efeitos dos fármacos , Linfócitos T/fisiologia , Benzimidazóis/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Citocinas/biossíntese , Humanos , Células Jurkat , Sistema de Sinalização das MAP Quinases , Tetrazóis/farmacologia
10.
Scand J Immunol ; 88(4): e12711, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30270447

RESUMO

In this study, the frequency and function of CD4+CD25+CD45RA+ regulatory T cells (Treg) and intracellular IL-2 signalling molecules in patients with type 2 diabetes mellitus (T2DM) were investigated. Tregs and responder T cells (Tresp, CD4+CD25- T cells) were sorted and suppression assays were performed using flow cytometry. Phosphorylation of signal transducer and activator of transcription-5 (pSTAT5) were assessed using flow cytometry. Gene expression of FOXP3 was performed with the SYBR green Real Time PCR method. Production of IL-2 from cultured cells was assessed using ELISA. We observed a functional defect of CD4+CD25+CD45RA+ Tregs in T2DM patients with higher proliferation of Tresp cells, in response to anti-CD3 and anti CD28 stimulation in the presence of Tregs in vitro. The results showed that the proliferation of Tresps in the absence of Treg cells was higher in T2DM patients than in healthy controls. Decreased FOXP3 mRNA expression and pSTAT5 were observed within the Tregs of the patients, whereas the level of secreted IL-2 from PBMCs culture was not statically different between T2DM patients and healthy individuals. Changes in intracellular IL-2 pathways and FOXP3 gene expression may contribute to the defect of Tregs in T2DM patients. These findings indicating that the purified CD4+CD25+CD45RA+ Treg cells have reduced functional capacity together with impaired IL-2 pathway in T2DM, and the Tregs could be used for a potential novel therapeutic target.


Assuntos
Diabetes Mellitus Tipo 2/imunologia , Subunidade alfa de Receptor de Interleucina-2/metabolismo , Interleucina-2/biossíntese , Linfócitos T Reguladores/imunologia , Adulto , Antígenos CD4/metabolismo , Feminino , Fatores de Transcrição Forkhead/biossíntese , Fatores de Transcrição Forkhead/genética , Humanos , Interleucina-2/metabolismo , Antígenos Comuns de Leucócito/metabolismo , Masculino , Pessoa de Meia-Idade , Fosforilação , RNA Mensageiro/genética , Fator de Transcrição STAT5/metabolismo , Proteínas Supressoras de Tumor/metabolismo
11.
J Int Med Res ; 46(9): 3938-3947, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29976100

RESUMO

Objective Alcohol is a hypnotic that modifies immune function, specifically the cytokines interferon gamma (IFN-γ) and interleukin 2 (IL-2). We evaluated the association between unscheduled napping and acute alcohol-induced augmentation of IFN-γ and IL-2 expression. Methods In this prospective, observational pilot study, volunteers completed questionnaires on sleep quality, alcohol use, and hangover characteristics. Actigraph recordings began three nights before and continued for four nights after study initiation. Napping was recorded by actigraphy and self-reporting. A weight-based dose of 100-proof vodka was consumed, and the blood alcohol content (BAC) and phytohemagglutinin-M stimulated cytokine level were measured before and 20 minutes, 2 hours, and 5 hours after binge consumption. Results Ten healthy volunteers participated (mean age, 34.4 ± 2.3 years; mean body mass index, 23.9 ± 4.6 kg/m2; 60% female). The mean 20-minute BAC was 137.7 ± 40.7 mg/dL. Seven participants took an unscheduled nap. The ex vivo IFN-γ and IL-2 levels significantly increased at all time points after binge consumption in the nappers, but not in the non-nappers. Conclusion Augmented IFN-γ and IL-2 levels are associated with unscheduled napping after binge alcohol consumption. Further studies are needed to clarify the associations among alcohol consumption, sleep disruption, and inflammatory mediators.


Assuntos
Consumo de Bebidas Alcoólicas/imunologia , Bebedeira/imunologia , Inflamação/imunologia , Interferon gama/biossíntese , Interleucina-2/biossíntese , Sono/imunologia , Actigrafia , Adulto , Consumo de Bebidas Alcoólicas/efeitos adversos , Bebedeira/complicações , Feminino , Inquéritos Epidemiológicos , Voluntários Saudáveis , Humanos , Imunidade Inata/imunologia , Masculino , Projetos Piloto , Estudos Prospectivos
12.
BMC Anesthesiol ; 18(1): 87, 2018 07 18.
Artigo em Inglês | MEDLINE | ID: mdl-30021515

RESUMO

BACKGROUND: Interleukin 2 (IL-2) influences the development and severity of pain due to its antinociceptive and immunomodulatory effects. Its production is influenced by the increased expression of c-Cbl (Casitas B-lineage lymphoma proto-oncogene) and Cbl-b E3 ubiquitin ligases. We evaluated the effects on IL-2-mediated changes in c-Cbl and Cbl-b expression in a rat model of chronic neuropathic pain. METHODS: Peripheral neuropathy was induced in adult male Sprague-Dawley rats weighing 250-300 g by chronic spinal nerve ligation. Half of the spinal cord ipsilateral to the nerve injury was harvested at 1, 3, and 6 weeks, and the expression levels of IL-2, c-Cbl, Cbl-b, phospholipase C-γ1 (PLC-γ1), ZAP70, and protein kinase Cθ (PKCθ), as well as ubiquitin conjugation, were evaluated. RESULTS: Total IL-2 mRNA levels were significantly decreased at 3 and 6 weeks after nerve injury compared to those in sham-operated rats. The mRNA levels of c-Cbl and Cbl-b, as well as the level of ubiquitin conjugation, were significantly increased at 3 and 6 weeks. In contrast, the levels of phosphorylated ZAP70 and PLC-γ1 were decreased at 3 and 6 weeks after spinal nerve ligation. Ubiquitination of PLC-γ1 and PKCθ was increased at 3 and 6 weeks. CONCLUSIONS: Our results suggest that ubiquitin and the E3 ubiquitin ligases c-Cbl and Cbl-b function as neuroimmune modulators in the subacute phase of neuropathic pain after nerve injury.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/biossíntese , Interleucina-2/biossíntese , Doenças do Sistema Nervoso Periférico/metabolismo , Proteínas Proto-Oncogênicas c-cbl/biossíntese , Ubiquitina-Proteína Ligases/biossíntese , Animais , Ligadura , Masculino , Fosfolipase C gama/biossíntese , Proteína Quinase C-theta/biossíntese , Ratos , Medula Espinal/metabolismo , Nervos Espinhais/lesões , Ubiquitina/metabolismo , Ubiquitinação , Proteína-Tirosina Quinase ZAP-70/biossíntese
13.
J Int Med Res ; 46(7): 2792-2802, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29938552

RESUMO

Objective To evaluate how isoflurane affects T-cell function by assaying interleukin (IL)-2 production and the expression of two Casitas B-lineage lymphoma (Cbl) family proto-oncogenes (c-Cbl and Cbl-b) in rat peripheral blood mononuclear cells (PBMCs). Methods Adult male Sprague-Dawley rats were randomly allocated to those that underwent blood collection after brief isoflurane anesthesia (control group), immediately after 4 hours of isoflurane general anesthesia (4I group), and 1 day after 4 hours of isoflurane general anesthesia (1D 4I group). IL-1, IL-2, and IL-6 mRNA levels and c-Cbl and Cbl-b levels in PBMCs were determined by polymerase chain reaction. Ubiquitination of protein kinase Cθ (PKCθ) and phospholipase C-γ1 (PLC-γ1) in PBMCs was assessed by immunoprecipitation. Results The IL-2 mRNA level in rat PBMCs was significantly lower in the 4I and 1D 4I groups than in the control group. c-Cbl, Cbl-b, and ubiquitin expression was significantly increased and zeta-chain-associated protein kinase 70, PLC-γ1, and PKCθ protein levels were significantly decreased in the 4I group. Ubiquitination of PLC-γ1 and PKCθ was significantly increased in the 4I group. Conclusion Isoflurane influences ubiquitin, c-Cbl, and Cbl-b expression in rat PBMCs, indicating suppression of receptor tyrosine kinase signaling pathways. These results suggest that isoflurane suppresses T-cell function.


Assuntos
Anestésicos Inalatórios/farmacologia , Interleucina-2/biossíntese , Isoflurano/farmacologia , Leucócitos Mononucleares/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-cbl/biossíntese , Linfócitos T/efeitos dos fármacos , Proteínas Adaptadoras de Transdução de Sinal/biossíntese , Animais , Leucócitos Mononucleares/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Linfócitos T/metabolismo , Ubiquitina/metabolismo
14.
Mol Immunol ; 101: 167-175, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29944986

RESUMO

Liver sinusoidal endothelial cells (LSECs) are organ resident APCs capable of antigen presentation and subsequent tolerization of T cells under physiological conditions. In this study, we investigated whether LSEC pretreatment with NOD-like receptor (NLR) agonists can switch the cells from a tolerogenic to an immunogenic state and promote the development of T cell immunity. LSECs constitutively express NOD1, NOD2 and RIPK2. Stimulation of LSECs with DAP induced the activation of NF-κB and MAP kinases and upregulated the expression of chemokines (CXCL2/9, CCL2/7/8) and cytokines (IFN-γ, TNF-α and IL-2). Pretreatment of LSECs with DAP induced significantly increased IFN-γ and IL-2-production by HBV-stimulated CD8+ T cells primed by DAP-treated LSECs. Consistently, a significant reduction in the HBV DNA and HBsAg level occurred in mice receiving T cells primed by DAP-treated LSECs. MDP stimulation had no impact on LSECs or HBV-stimulated CD8+ T cells primed with MDP-treated LSECs except for the upregulation of PD-L1. DAP stimulation in vitro could promote LSEC maturation and activate HBV-specific T cell responses. These results are of particular relevance for the regulation of the local innate immune response against HBV infections.


Assuntos
Diferenciação Celular , Células Endoteliais/metabolismo , Imunidade Celular , Fígado/citologia , Proteína Adaptadora de Sinalização NOD1/metabolismo , Linfócitos T/citologia , Linfócitos T/imunologia , Acetilmuramil-Alanil-Isoglutamina/farmacologia , Animais , Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/metabolismo , Diferenciação Celular/efeitos dos fármacos , Ácido Diaminopimélico/farmacologia , Células Endoteliais/efeitos dos fármacos , Vírus da Hepatite B/efeitos dos fármacos , Vírus da Hepatite B/fisiologia , Imunidade Celular/efeitos dos fármacos , Interleucina-2/biossíntese , Ligantes , Camundongos Endogâmicos C57BL , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Proteína Serina-Treonina Quinase 2 de Interação com Receptor/metabolismo , Regulação para Cima/efeitos dos fármacos
15.
Biochem Biophys Res Commun ; 502(2): 226-231, 2018 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-29792863

RESUMO

Human infections by type B influenza virus constitute about 25% of all influenza cases. The viral hemagglutinin is comprised of two subunits, HA1 and HA2. While HA1 is constantly evolving in an unpredictable fashion, the HA2 subunit is highly conserved, making it a potential candidate for a universal vaccine. However, immunodominant epitopes in the HA2 subunit remain largely unknown. To delineate MHC Class I epitopes, we first identified 9-mer H-2Kd-restricted CD8 T cell epitopes in the HA2 domain by in silico analyses, followed by evaluating the immunodominance of these peptides in mice challenged with the virus. Of three peptides selected through in silico analysis, the universally conserved peptide, YYSTAASSL (B/HA2-190), possessed the highest predicted binding affinity to MHC Class I and was most effective in inducing IL-2 and TNF-α in mouse splenocytes. Importantly, the peptide demonstrated best capability of stimulating peptide-specific ex-vivo cytotoxicity against target cells. Taken together, this finding would be of value for assessment of cell-mediated immune responses elicited by vaccines based on the highly conserved HA2 stalk domain.


Assuntos
Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Vírus da Influenza B/imunologia , Animais , Antígenos CD8/química , Simulação por Computador , Feminino , Antígenos H-2/imunologia , Glicoproteínas de Hemaglutininação de Vírus da Influenza/química , Humanos , Imunidade Celular , Epitopos Imunodominantes/química , Vírus da Influenza B/química , Vacinas contra Influenza/química , Vacinas contra Influenza/imunologia , Influenza Humana/imunologia , Influenza Humana/virologia , Interleucina-2/biossíntese , Camundongos , Camundongos Endogâmicos DBA , Modelos Imunológicos , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/virologia , Subunidades Proteicas , Linfócitos T Citotóxicos/imunologia , Fator de Necrose Tumoral alfa/biossíntese
16.
Nat Immunol ; 19(5): 497-507, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29662170

RESUMO

The transcription factor c-Maf induces the anti-inflammatory cytokine IL-10 in CD4+ T cells in vitro. However, the global effects of c-Maf on diverse immune responses in vivo are unknown. Here we found that c-Maf regulated IL-10 production in CD4+ T cells in disease models involving the TH1 subset of helper T cells (malaria), TH2 cells (allergy) and TH17 cells (autoimmunity) in vivo. Although mice with c-Maf deficiency targeted to T cells showed greater pathology in TH1 and TH2 responses, TH17 cell-mediated pathology was reduced in this context, with an accompanying decrease in TH17 cells and increase in Foxp3+ regulatory T cells. Bivariate genomic footprinting elucidated the c-Maf transcription-factor network, including enhanced activity of NFAT; this led to the identification and validation of c-Maf as a negative regulator of IL-2. The decreased expression of the gene encoding the transcription factor RORγt (Rorc) that resulted from c-Maf deficiency was dependent on IL-2, which explained the in vivo observations. Thus, c-Maf is a positive and negative regulator of the expression of cytokine-encoding genes, with context-specific effects that allow each immune response to occur in a controlled yet effective manner.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Regulação da Expressão Gênica/imunologia , Redes Reguladoras de Genes/imunologia , Interleucina-2/biossíntese , Proteínas Proto-Oncogênicas c-maf/imunologia , Animais , Interleucina-2/imunologia , Camundongos
17.
Anticancer Res ; 38(3): 1327-1333, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29491056

RESUMO

BACKGROUND/AIM: ß-Glucans are well-established immunomodulators with strong effects across all immune reactions. Due to the extensive amount of studies, glucans are steadily progressing from a non-specific immunomodulator to a licensed drug. However, direct comparisons of higher numbers of different glucans are rare. MATERIALS AND METHODS: In this study, we used 16 different glucans isolated from yeasts, mushroom, algae, and oat and compared their effects on phagocytosis, IL-2 production, antibody secretion, and inhibition of three experimental cancer models. RESULTS: Our results showed significant differences among tested glucans, showing that despite the fact that glucans in general have strong stimulating effects on most aspects of the immune system, it is necessary to choose the right glucan. CONCLUSION: Based on our studies, we can conclude that highly purified and active glucans have significant pleiotropic effects.


Assuntos
Carcinoma Pulmonar de Lewis/tratamento farmacológico , Modelos Animais de Doenças , Neoplasias Mamárias Experimentais/tratamento farmacológico , beta-Glucanas/farmacologia , Agaricales/química , Animais , Formação de Anticorpos/efeitos dos fármacos , Avena/química , Carcinoma Pulmonar de Lewis/patologia , Linhagem Celular Tumoral , Feminino , Humanos , Interleucina-2/biossíntese , Neoplasias Mamárias Experimentais/patologia , Camundongos Endogâmicos BALB C , Fagocitose/efeitos dos fármacos , Baço/citologia , Baço/efeitos dos fármacos , Baço/metabolismo , Leveduras/química , beta-Glucanas/isolamento & purificação
18.
Georgian Med News ; (Issue): 141-150, 2018 Feb.
Artigo em Russo | MEDLINE | ID: mdl-29578440

RESUMO

The objective of this researsh is to study the effects of Eminium Regelii phytopreparation (ERP) on immune status and free radical oxidation in the tissues of the adrenal glands and immunocompetent organs after combined exposure to 6 Gy dose of gamma irradiation and coal dust (remote period). The study was realized on 30 white laboratory male rats of the Wistar line, weighing 240±20g, that were divided into equal 3 groups: I group - intact, ІІ group - were exposured to combined effects of coal dust and gamma irradiation, III group - were exposured to combined effects and in parallel taking phytopreparation Eminium Regel. The animals of II and III groups were irradiated 90 days prior to the study at the TERAGAM 60Co radiotherapy unit ("ISOTREND spol. S.r.o.", Czech Republic) in dose of 6 Gy once. Experimental animals received phytopreparation of ER 2.5 mg/kg per day on calculate of body mass for 14 days. The results of the conducted studies showed that in the long-term period after the actions of the sublethal dose of gamma radiation and coal dust, significant changes were revealed that were characterized by a decrease in immunological reactivity, increased lipoperoxidation and inhibition of antioxidant defense activity of the organism. After exposure to ER, oxidative stress was alleviated, sufficient restoration of antioxidant protection and immune system indices, which were disrupted by the combined effects of a single high dose of radiation and a prolonged three-month inhalation of coal dust.


Assuntos
Antioxidantes/farmacologia , Araceae/química , Carvão Mineral/toxicidade , Raios gama/efeitos adversos , Material Particulado/antagonistas & inibidores , Protetores contra Radiação/farmacologia , Glândulas Suprarrenais/efeitos dos fármacos , Glândulas Suprarrenais/imunologia , Glândulas Suprarrenais/efeitos da radiação , Animais , Antígenos CD/biossíntese , Antígenos CD/imunologia , Antioxidantes/isolamento & purificação , Linfócitos B/efeitos dos fármacos , Linfócitos B/imunologia , Linfócitos B/efeitos da radiação , Esquema de Medicação , Poeira/análise , Interferon gama/biossíntese , Interferon gama/imunologia , Interleucina-2/biossíntese , Interleucina-2/imunologia , Interleucina-6/biossíntese , Interleucina-6/imunologia , Peroxidação de Lipídeos/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos da radiação , Fígado/efeitos dos fármacos , Fígado/imunologia , Fígado/efeitos da radiação , Linfonodos/efeitos dos fármacos , Linfonodos/imunologia , Linfonodos/efeitos da radiação , Masculino , Material Particulado/toxicidade , Extratos Vegetais/química , Protetores contra Radiação/isolamento & purificação , Ratos , Ratos Wistar , Baço/efeitos dos fármacos , Baço/imunologia , Baço/efeitos da radiação , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Linfócitos T/efeitos da radiação , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/imunologia
19.
Microb Pathog ; 117: 206-218, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29476787

RESUMO

Here, we studied the in vivo expression of Th1 (IL2 and IFN gamma) and Th2 (IL4 and IL10) - cytokines and antiviral molecules - IRF3 and ISG15 in peripheral blood mononuclear cells in relation to antigen and antibody dynamics under Peste des petits ruminants virus (PPRV) vaccination, infection and challenge in both sheep and goats. Vaccinated goats were seropositive by 9 days post vaccination (dpv) while in sheep idiosyncratic response was observed between 9 and 14 dpv for different animals. Expression of PPRV N gene was not detected in PBMCs of vaccinated and vaccinated challenged groups of both species, but was detected in unvaccinated infected PBMCs at 9 and 14 days post infection. The higher viral load at 9 dpi coincided with the peak clinical signs of the disease. The peak in viral replication at 9 dpi correlated with significant expression of antiviral molecules IRF3, ISG15 and IFN gamma in both the species. With the progression of disease, the decrease in N gene expression also correlated with the decrease in expression of IRF3, ISG15 and IFN gamma. In the unvaccinated infected animals ISG15, IRF3, IFN gamma and IL10 expression was higher than vaccinated animals. The IFN gamma expression predominated over IL4 in both vaccinated and infected animals with the infected exhibiting a stronger Th1 response. The persistent upregulation of this antiviral molecular signature - ISG15 and IRF3 even after 2 weeks post vaccination, presumably reflects the ongoing stimulation of innate immune cells.


Assuntos
Citocinas/biossíntese , Regulação da Expressão Gênica/imunologia , Peste dos Pequenos Ruminantes/imunologia , Vírus da Peste dos Pequenos Ruminantes/imunologia , Tropismo/imunologia , Vacinação/veterinária , Vacinas Virais/imunologia , Eliminação de Partículas Virais/imunologia , Animais , Anticorpos Antivirais/sangue , Antígenos Virais/sangue , Antivirais/farmacologia , Citocinas/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/genética , Genes Virais/genética , Doenças das Cabras/imunologia , Doenças das Cabras/prevenção & controle , Doenças das Cabras/virologia , Cabras , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia , Fator Regulador 3 de Interferon/biossíntese , Fator Regulador 3 de Interferon/genética , Interferon gama/biossíntese , Interferon gama/genética , Interleucina-10/biossíntese , Interleucina-10/genética , Interleucina-2/biossíntese , Interleucina-2/genética , Interleucina-4/biossíntese , Interleucina-4/genética , Cinética , Leucócitos Mononucleares/imunologia , Peste dos Pequenos Ruminantes/patologia , Peste dos Pequenos Ruminantes/prevenção & controle , Peste dos Pequenos Ruminantes/virologia , Vírus da Peste dos Pequenos Ruminantes/genética , Vírus da Peste dos Pequenos Ruminantes/patogenicidade , Ruminantes/imunologia , Ovinos , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/prevenção & controle , Doenças dos Ovinos/virologia , Fatores de Tempo , Vacinas Atenuadas/imunologia , Carga Viral , Replicação Viral
20.
Biotechnol Bioeng ; 115(3): 565-576, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29178403

RESUMO

Anti-CD20 recombinant antibodies are among the most promising therapeutics for the treatment of B-cell malignancies such as non-Hodgkin lymphomas. We recently demonstrated that an immunocytokine (2B8-Fc-hIL2), obtained by fusing an anti-CD20 scFv-Fc antibody derived from C2B8 mAb (rituximab) to the human interleukin 2 (hIL-2), can be efficiently produced in Nicotiana benthamiana plants. The purified immunocytokine (IC) bearing a typical plant protein N-glycosylation profile showed a CD20 binding activity comparable to that of rituximab and was efficient in eliciting antibody-dependent cell-mediated cytotoxicity (ADCC) of human PBMC against Daudi cells, indicating its fuctional integrity. In this work, the immunocytokine devoid of the typical xylose/fucose N-glycosylation plant signature (IC-ΔXF) and the corresponding scFv-Fc-ΔXF antibody not fused to the cytokine, were obtained in a glyco-engineered ΔXylT/FucT N. benthamiana line. Purification yields from agroinfiltrated plants amounted to 20-35 mg/kg of leaf fresh weight. When assayed for interaction with FcγRI and FcγRIIIa, IC-ΔXF exhibited significantly enhanced binding affinities if compared to the counterpart bearing the typical plant protein N-glycosylation profile (IC) and to rituximab. The glyco-engineered recombinant molecules also exhibited a strongly improved ADCC and complement-dependent cytotoxicity (CDC). Notably, our results demonstrate a reduced C1q binding of xylose/fucose carrying IC and scFv-Fc compared to versions that lack these sugar moieties. These results demonstrate that specific N-glycosylation alterations in recombinant products can dramatically affect the effector functions of the immunocytokine, resulting in an overall improvement of the biological functions and consequently of the therapeutic potential.


Assuntos
Interleucina-2 , Leucócitos Mononucleares/metabolismo , Plantas Geneticamente Modificadas , Polissacarídeos , Proteínas Recombinantes de Fusão , Anticorpos de Cadeia Única , Tabaco , Humanos , Interleucina-2/biossíntese , Interleucina-2/química , Interleucina-2/genética , Interleucina-2/farmacologia , Leucócitos Mononucleares/citologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Polissacarídeos/biossíntese , Polissacarídeos/genética , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Engenharia de Proteínas , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteínas Recombinantes de Fusão/farmacologia , Anticorpos de Cadeia Única/biossíntese , Anticorpos de Cadeia Única/genética , Anticorpos de Cadeia Única/isolamento & purificação , Anticorpos de Cadeia Única/farmacologia , Tabaco/genética , Tabaco/metabolismo
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