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1.
Plast Reconstr Surg ; 143(6): 1215e-1223e, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31136482

RESUMO

BACKGROUND: Large calvarial defects represent a major reconstructive challenge, as they do not heal spontaneously. Infection causes inflammation and scarring, further reducing the healing capacity of the calvaria. Bone morphogenetic protein-2 (BMP2) has been shown to stimulate osteogenesis but has significant side effects in high doses. BMP2 has not been tested in combination with antiinflammatory cytokines such as interleukin-10. METHODS: Sixteen New Zealand White rabbits underwent 15 × 15-mm flap calvarectomies. The flap was incubated in Staphylococcus aureus and replaced, and infection and scarring were allowed to develop. The flap was subsequently removed and the wound débrided. A 15 × 15-mm square of acellular dermal matrix biopatterned with low-dose BMP2, interleukin-10, or a combination was implanted. Computed tomographic scans were taken over 42 days. Rabbits were then killed and histology was performed. RESULTS: Defects treated with BMP2 showed significantly (p < 0.05) greater osseous regeneration than untreated controls. Interleukin-10 did not significantly augment the healing achieved with BMP2, and interleukin-10 alone did not significantly increase healing compared with controls. Histology showed evidence of bone formation in defects treated with BMP2. Untreated controls and defects treated with interleukin-10 alone showed only fibrous tissue in the defect site. CONCLUSIONS: Low-dose BMP2 delivered directly to the scarred calvarial defect augments bony healing. Interleukin-10 at the dose applied did not significantly augment healing alone or in combination with BMP2. Healing had not finished at 42 days and analysis at later time points or the use of higher doses of BMP2 may yield greater healing.


Assuntos
Proteína Morfogenética Óssea 2/farmacologia , Regeneração Óssea/efeitos dos fármacos , Interleucina-10/farmacologia , Crânio/fisiologia , Animais , Proteína Morfogenética Óssea 2/administração & dosagem , Cicatriz/tratamento farmacológico , Combinação de Medicamentos , Interleucina-10/administração & dosagem , Masculino , Coelhos , Crânio/efeitos dos fármacos , Crânio/cirurgia , Infecções Estafilocócicas/fisiopatologia , Staphylococcus aureus , Retalhos Cirúrgicos , Tomografia Computadorizada por Raios X
2.
Vet Microbiol ; 231: 139-146, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30955801

RESUMO

The recent emergence of highly pathogenic porcine reproductive and respiratory syndrome virus 1 (PRRSV-1) strains has caused severe economic losses. The biological elements defining virulence and pathogenicity are still unclear. In vitro characteristics using natural target cells of PRRSV provide important information to understand the basis of virulence at the cellular level, and provide a mean to reduce animal experimentations to achieve this goal. Here, we compared PRRSV strains from two geographically different regions, with varying in vivo characteristics, in terms of their interactions with monocyte-derived macrophages (MDMs). The strains included Lena and BOR59 from Belarus, and ILI6 from Russia, as well as PR11 and PR40, both from Italy. As a reference, we used a cell culture-adapted version of Lelystad, LVP. MDMs were pre-treated with IFNγ, IL-4 or IFNß, in order to understand responses in polarized and antiviral MDMs. In general, independent of the geographical origin, the strains with high virulence infected a higher percentage of MDMs and replicated to higher titers. These virulence-dependent differences were most pronounced when the MDMs had been treated with IFNß. Differentiation between intermediate and low virulent PRRSV was difficult, due to variations between different experiments, but LVP differed clearly from all field strains. IFNα and IL-10 were not detected in any experiment, but PR40 induced TNF and IL-1ß. Taken together, these results validate the MDM model to understand pathogenicity factors of PRRSV and confirm the importance of the escape from type I and II IFN-mediated effects for PRRSV virulence.


Assuntos
Macrófagos/virologia , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/patogenicidade , Animais , Interferon-alfa/farmacologia , Interferon beta/farmacologia , Interleucina-10/farmacologia , Itália , Macrófagos/efeitos dos fármacos , Vírus da Síndrome Respiratória e Reprodutiva Suína/classificação , República de Belarus , Federação Russa , Suínos , Virulência
3.
Med Sci Monit ; 25: 2923-2934, 2019 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-31005957

RESUMO

BACKGROUND Rheumatoid arthritis model (CIA) rats were treated by tail vein injection of IL-10-modified bone marrow mesenchymal stem cells (BMSCs) to investigate its feasibility and intrinsic molecular mechanism. MATERIAL AND METHODS The CIA rat model was established by induction type II collagen, and IL-10-modified BMSCs was established by transfecting BMSCs with adenovirus. IL-10-modified BMSCs were used to treat the CIA rats. The therapeutic effect was evaluated by measuring the changes in body weight, ankle swelling, and forced swimming time, as well as observation of synovial hyperplasia and cartilage tissue repair by HE staining. Western blot analysis and ELISA were used to detect gene expression. RESULTS After 4 weeks and 8 weeks of treatment with IL10-BMSCs, the body weight, swelling value, resting time, and forced swimming struggle time of CIA rats were significantly higher than those of BMSCs-treated and -untreated CIA rats (P<0.05). Compared to BMSCs-treated CIA model rats, after treatment with IL10-BMSCs, the repair rate of osteoarticular cartilage was higher and the inhibition of synovial proliferation was better, and serum IL-17, IL-1ß, and TNF-alpha levels were lower. We found that the protein level of SIRT1 in peripheral blood mononuclear cells was lower, the protein level in spleen was higher, and phosphorylation of p65 protein in peripheral blood mononuclear cells was reduced. CONCLUSIONS The efficacy of tail vein injection of IL-10-modified BMSCs in treatment of CIA rats was superior to that of BMSCs alone, which may be related to the more pronounced suppression of IL-10-modified BMSCs in peripheral blood inflammation and spleen immune response.


Assuntos
Artrite Reumatoide/tratamento farmacológico , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/fisiologia , Animais , Artrite Experimental/tratamento farmacológico , Artrite Reumatoide/induzido quimicamente , Medula Óssea , Células da Medula Óssea , Cartilagem/efeitos dos fármacos , Colágeno Tipo II/farmacologia , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Mediadores da Inflamação/metabolismo , Interleucina-10/metabolismo , Interleucina-10/farmacologia , Interleucina-17/sangue , Interleucina-17/metabolismo , Interleucina-1beta/sangue , Interleucina-1beta/metabolismo , Leucócitos Mononucleares/metabolismo , Células-Tronco Mesenquimais/patologia , Ratos , Ratos Wistar , Sirtuína 1/metabolismo , Fator de Necrose Tumoral alfa/sangue , Fator de Necrose Tumoral alfa/metabolismo
4.
Neurosci Lett ; 703: 132-138, 2019 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-30904573

RESUMO

Many trigeminal neuropathic pain patients suffer severe chronic pain. The neuropathic pain might be related with cross-excitation of the neighboring neurons and satellite glial cells (SGCs) in the sensory ganglia and increasing the pain signals from the peripheral tissue to the central nervous system. We induced trigeminal neuropathic pain by infraorbital nerve constriction injury (IONC) in Sprague-Dawley rats. We tested cytokine (CXCL2 and IL-10) levels in trigeminal ganglia (TGs) after trigeminal neuropathic pain induction, and the effect of direct injection of the anti-CXCL2 and recombinant IL-10 into TG. We found that IONC induced pain behavior. Additionally, IONC induced satellite glial cell activation in TG and cytokine levels of TGs were changed after IONC. CXCL2 levels increased on day 1 of neuropathic pain induction and decreased gradually, with IL-10 levels showing the opposite trend. Recombinant IL-10 or anti-CXCL2 injection into TG decreased pain behavior. Our results show that IL-10 or anti-CXCL2 are therapy options for neuropathic pain.


Assuntos
Quimiocina CXCL2/metabolismo , Interleucina-10/metabolismo , Neuralgia/metabolismo , Gânglio Trigeminal/metabolismo , Animais , Anticorpos/farmacologia , Quimiocina CXCL2/imunologia , Constrição Patológica , Interleucina-10/farmacologia , Masculino , Neuralgia/fisiopatologia , Medição da Dor , Traumatismos dos Nervos Periféricos/fisiopatologia , Ratos Sprague-Dawley , Proteínas Recombinantes/farmacologia
5.
J Neuroinflammation ; 16(1): 66, 2019 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-30922332

RESUMO

BACKGROUND: Microglia are important for secreting chemical mediators of inflammatory responses in the central nervous system. Interleukin (IL)-10 and IL-1ß secreted by glial cells support neuronal functions, but the related mechanisms remain vague. Our goal was to demonstrate the efficacy of IL-10 in suppressing IL-1ß and in inflammasome activation in mice with epileptic seizure based on an epileptic-seizure mouse model. METHODS: In this study, mice in which epileptic seizures were induced by administering picrotoxin (PTX) were used as a case group, and mice injected with saline were employed as the control group. The expression of nucleic acids, cytokines, or signaling pathways was detected by reverse transcription-polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), flow cytometry, and Western blotting. RESULTS: Our results demonstrated that IL-10 inhibits IL-1ß production through two distinct mechanisms: (1) Treatment with lipopolysaccharides (LPS) results in IL-10 overexpression in microglia and reduced NLRP3 inflammasome activity, thus inhibiting caspase-1-related IL-1ß maturation; (2) next, autocrine IL-10 was found to subsequently promote signal transducer and activator of transcription-3 (STAT-3), reducing amounts of pro-IL-1ß. CONCLUSIONS: Our results indicate that IL-10 is potentially effective in the treatment of inflammation encephalopathy, and suggest the potential usefulness of IL-10 for treating autoimmune or inflammatory ailments.


Assuntos
Interleucina-10/farmacologia , Interleucina-1beta/metabolismo , Microglia/metabolismo , Convulsões/patologia , Animais , Encéfalo/patologia , Células Cultivadas , Convulsivantes/toxicidade , Citocinas/genética , Citocinas/metabolismo , Modelos Animais de Doenças , Regulação da Expressão Gênica/efeitos dos fármacos , Lipopolissacarídeos/toxicidade , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microglia/efeitos dos fármacos , Ácidos Nucleicos/genética , Ácidos Nucleicos/metabolismo , Picrotoxina/toxicidade , RNA Interferente Pequeno/farmacologia , Fator de Transcrição STAT3/metabolismo , Convulsões/induzido quimicamente , Convulsões/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética
6.
Int J Mol Sci ; 20(3)2019 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-30759730

RESUMO

The association between osteoarthritis (OA), obesity and metabolic syndrome suggests an interrelation between OA and diabetes mellitus (DM). Little is known about the role of anti-inflammatory cytokine interleukin (IL)-10 in the interrelation between OA and DM. Hence, the effects of IL-10 under hyperglycemia (HG) and hyperinsulinemia (HI) in human articular chondrocytes (hAC) and chondrosarcoma cell line Okayama University Medical School (OUMS)-27 were examined. HAC and OUMS-27, cultured in normoglycemic (NG) and HG conditions were stimulated with insulin and/or IL-10. Cell survival, metabolic activity, proliferation and extracellular matrix (ECM) synthesis were immunocytochemically examined. No significant differences in vitality of hAC neither in pure NG (NGw/o) nor HG (HGw/o) conditions were found. Applying HI and/or IL-10 in both conditions reduced significantly the vitality of hAC but not of OUMS-27. HG impaired significantly hAC metabolism. When combined with HI + IL-10 or IL-10 alone it decreased also significantly hAC proliferation compared to NGw/o. In OUMS-27 it induced only a trend of impaired proliferation compared to NGw/o. hAC but not OUMS-27 reduced significantly their collagen type (col) I, SOX9 and proteoglycan (PG) synthesis in HG combined with HI +/- IL-10 compared to NGw/o. IL-10 could not moderate HI and HG effects. In contrast to hAC OUMS-27 showed limited sensitivity as DM model.


Assuntos
Diabetes Mellitus/tratamento farmacológico , Interleucina-10/farmacologia , Osteoartrite/tratamento farmacológico , Idoso , Células Cultivadas , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Diabetes Mellitus/metabolismo , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/metabolismo , Feminino , Humanos , Hiperglicemia/tratamento farmacológico , Hiperglicemia/metabolismo , Hiperinsulinismo/tratamento farmacológico , Hiperinsulinismo/metabolismo , Interleucina-10/metabolismo , Masculino , Síndrome Metabólica/tratamento farmacológico , Síndrome Metabólica/metabolismo , Osteoartrite/metabolismo
7.
Methods Mol Biol ; 1960: 101-112, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30798525

RESUMO

Macrophages are innate immune cells, which have important roles in the inflammatory response to infections or tissue injury, and have an equally important role in the resolution of inflammation. Macrophages play a key part in directing the innate immune response and subsequent adaptive immune response. They can acquire a variety of distinct but also overlapping activation states, depending on the local microenvironment, in order to perform these functions. Stimuli, such as IFNγ and LPS, can promote an inflammatory activation state, which is associated with the production of reactive oxygen species, and pro-inflammatory cytokines and chemokines. Immune complexes and LPS can promote an anti-inflammatory activation state to prevent damage to the host, which is associated with the production of high levels of the anti-inflammatory cytokine IL-10 and low levels of pro-inflammatory cytokines. Wound-healing macrophages can be activated by IL-4 or IL-13 and have roles in tissue remodeling and the resolution of inflammation. Macrophages are present in nearly every tissue of the body and are important for maintaining homeostasis, but their dysfunction can also lead to diseases, such as inflammatory bowel disease. To study the role macrophages play in a complex in vivo environment, depletion and reconstitution experiments can be utilized. Clodronate liposomes are an effective and versatile way to deplete macrophages in vivo; they can allow selective depletion from tissues of interest and can be used on transgenic mice. However, clodronate liposomes deplete all types of macrophages as well as dendritic cells, so other strategies are required in parallel to determine whether macrophages or macrophages of a particular activation state are required. Reconstitution of macrophages by adoptive transfer can be performed, with or without prior depletion, to further suggest that the observed effect is macrophage dependent. Macrophages activated ex vivo or macrophages from transgenic mice can be adoptively transferred during disease models to determine whether a specific protein or activation state affects disease outcome. Macrophage contribution to health and disease can be effectively studied using depletion with clodronate liposomes and by macrophage reconstitution, as demonstrated in this chapter.


Assuntos
Macrófagos/metabolismo , Animais , Ácido Clodrônico/metabolismo , Inflamação/metabolismo , Interferon gama/farmacologia , Interleucina-10/farmacologia , Interleucina-13/farmacologia , Interleucina-4/farmacologia , Lipopolissacarídeos/farmacologia , Lipossomos/metabolismo , Macrófagos/efeitos dos fármacos , Camundongos
8.
Cancer Immunol Immunother ; 68(3): 489-502, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30637473

RESUMO

Tumor-associated macrophages (TAMs) are of major importance in cancer-related immune suppression, and tumor infiltration by CD163pos TAMs is associated with poor outcome in most human cancers. Therefore, therapeutic strategies for reprogramming TAMs from a tumor-supporting (M2-like) phenotype towards a tumoricidal (M1-like) phenotype are of great interest. Activation of the transcription factor STAT3 within the tumor microenvironment is associated with worse prognosis, and STAT3 activation promotes the immunosuppressive phenotype of TAMs. Therefore, we aimed to develop a drug for inhibition of STAT3 specifically within human TAMs by targeting the endocytic CD163 scavenger receptor, which is highly expressed on TAMs. Here, we report the first data on a CD163-targeted STAT3-inhibitory drug consisting of corosolic acid (CA) packaged within long-circulating liposomes (LCLs), which are CD163-targeted by modification with monoclonal anti-CD163 antibodies (αCD163)-CA-LCL-αCD163. We show, that activation of STAT3 (by phosphorylation) was inhibited by CA-LCL-αCD163 specifically within CD163pos cells, with minor effect on CD163neg cells. Furthermore, CA-LCL-αCD163 inhibited STAT3-regulated gene expression of IL-10, and increased expression of TNFα, thus indicating a pro-inflammatory effect of the drug on human macrophages. This M1-like reprogramming at the mRNA level was confirmed by significantly elevated levels of pro-inflammatory cytokines (IFNγ, IL-12, TNFα, IL-2) in the culture medium. Since liposomes are attractive vehicles for novel anti-cancer drugs, and since direct TAM-targeting may decrease adverse effects of systemic inhibition of STAT3, the present results encourage future investigation of CA-LCL-αCD163 in the in vivo setting.


Assuntos
Macrófagos/fisiologia , Monócitos/fisiologia , Receptores de Superfície Celular/antagonistas & inibidores , Fator de Transcrição STAT3/antagonistas & inibidores , Triterpenos/administração & dosagem , Antígenos CD , Antígenos de Diferenciação Mielomonocítica , Células Cultivadas , Citocinas/biossíntese , Composição de Medicamentos , Humanos , Interleucina-10/farmacologia , Lipossomos , Macrófagos/efeitos dos fármacos , Monócitos/efeitos dos fármacos , Triterpenos/toxicidade
9.
Biochem Biophys Res Commun ; 509(4): 877-885, 2019 02 19.
Artigo em Inglês | MEDLINE | ID: mdl-30642634

RESUMO

Obesity is known to be induced by the accumulation of hypertrophy and hyperplasia of newly created fat in adipose tissues through differentiation of adipocyte precursor cells. Some cytokines are excessively produced in adipose tissues that can negatively regulate differentiation of adipocytes. Impaired adipogenesis is known to contribute to obesity-related diseases. Interleukin-10 (IL-10) is involved in the development of type 2 diabetes, insulin sensitivity, and immune response in obesity state. However, effects of IL-10 on adipogenesis remain unclear. The objective of this study was to determine the inhibitory effect of IL-10 on adipocyte differentiation and mechanisms involved in such effect. The effect of IL-10 on adipogenesis was analyzed by Western blot analysis, Oil Red O staining, qRT-PCR, and flow cytometry. We also examined the part of Wnt5a in adipogenesis using gene interfering technique. IL-10 suppressed lipid accumulation and adipocyte differentiation related gene expression. The inhibitory effect of IL-10 on the differentiation of adipocytes occurred at an early phase. IL-10 treatment caused a G0/G1 phase cell cycle arrest and altered expression levels of cell cycle proteins (CDK2, p21, and p27), thereby preventing re-entry into cell cycle. Additionally, IL-10 treatment reduced Wnt5a expression. Inhibition of Wnt5a by siRNA significantly attenuated lipid accumulation and expression of adipocyte differentiation-related genes. Taken together, these results indicate that IL-10 can inhibit the early phase of adipogenesis via suppressing Wnt5a signaling pathway in 3T3-L1 preadipocytes.


Assuntos
Adipócitos/metabolismo , Adipogenia/efeitos dos fármacos , Interleucina-10/farmacologia , Transdução de Sinais/fisiologia , Proteína Wnt-5a/metabolismo , Células 3T3-L1 , Adipócitos/citologia , Animais , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Camundongos
10.
Cancer Immunol Immunother ; 67(11): 1695-1707, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30128739

RESUMO

Tumor-mediated immunosuppression via regulatory T-cells is a key player among the various immune-escape mechanisms in multiple myeloma. We analyzed the generation, distribution, function and immunophenotype of CD8+CD28- regulatory T-cells in patients with multiple myeloma. Functionality of CD8+CD28- T-cells was assessed by immunological assays using ex vivo generated antigen-specific T-cells from patients with plasma cell dyscrasias and healthy donors. Detailed analysis of distribution, immunophenotype and cytotoxic potential of CD8+CD28- T-cells was performed by flow cytometry and ELISA. We found that the amount of CD8+CD28- T-cells was directly correlated with the suppression of antigen-specific T-cell responses in patients with plasma cell dyscrasia. Analyzing the CD8+CD28- T-cells in detail, increased numbers of these cells were observed in the bone marrow (i.e., tumor microenvironment) of patients with plasma cell dyscrasia. Furthermore, we identified the expression of lymphocyte function-associated antigen 1 (LFA-1) as a marker of immunosuppression and defined the CD8+CD28-CD57+LFA-1high population as the relevant immunosuppressive compartment. These regulatory T-cells act as immunosuppressors via soluble factors and incubation with IL-10 augmented their immunosuppressive capacity. The immunosuppressive regulatory network of IL-10 and the CD8+CD28-CD57+LFA-1high regulatory T-cells show unique characteristics and contribute to the tumor immune escape mechanism in patients with multiple myeloma.


Assuntos
Antígenos CD28/imunologia , Antígenos CD8/imunologia , Linfócitos T CD8-Positivos/imunologia , Interleucina-10/farmacologia , Mieloma Múltiplo/imunologia , Paraproteinemias/imunologia , Subpopulações de Linfócitos T/imunologia , Linfócitos T Reguladores/imunologia , Linfócitos T CD8-Positivos/efeitos dos fármacos , Estudos de Casos e Controles , Células Cultivadas , Humanos , Ativação Linfocitária , Mieloma Múltiplo/metabolismo , Mieloma Múltiplo/patologia , Paraproteinemias/metabolismo , Paraproteinemias/patologia , Subpopulações de Linfócitos T/efeitos dos fármacos , Linfócitos T Citotóxicos/efeitos dos fármacos , Linfócitos T Citotóxicos/imunologia , Linfócitos T Reguladores/efeitos dos fármacos
11.
Med Sci Monit ; 24: 4433-4439, 2018 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-29949812

RESUMO

BACKGROUND The aim of this study was to investigate the effects of TNF-α and IL-10 on the expression of ICAM-1 and CD31 in human coronary artery endothelial cells (HCAEC). MATERIAL AND METHODS HCAEC was treated with 0, 2.5 µg/l, 5 µg/l, and 10 µg/l of TNF-α for 2 h, 6 h, and 10 h, and with 0 µg/l, 10 µg/l, 100 µg/l, and 200 µg/l of IL-10 for 5 h, 10 h and 15 h, respectively. RNA inference of TNF-αR was performed with siRNA. Real-time PCR, Western blot analysis, and ELSA were used to detect the mRNA level and protein level of ICAM-1 and CD31. RESULTS TNF-α significantly increased the mRNA and protein expression of ICAM-1 (P<0.05), and 2.5 µg/l TNF-α had the most obvious effect. RNAi of TNF-aR reduced the induction of TNF-α on the mRNA and protein expression of ICAM-1 (P<0.05). TNF-α significantly decreased the CD31 in the supernatant (P<0.05), and 2.5 µg/l TNF-a had the most obvious effect. IL-10 significantly decreased the ICAM-1 protein level. IL-10 decreased the mRNA expression and the protein expression of CD31. The effect on mRNA was not significant (P>0.05), while the effect on the protein expression was significant (P<0.05). CONCLUSIONS TNF-α and IL-10 treatment can affect the expression of ICAM-1 and CD31 in HCAEC.


Assuntos
Vasos Coronários/citologia , Células Endoteliais/metabolismo , Molécula 1 de Adesão Intercelular/metabolismo , Interleucina-10/farmacologia , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Células Endoteliais/efeitos dos fármacos , Humanos
12.
PLoS One ; 13(5): e0197223, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29763436

RESUMO

Bandaging of limb wounds in horses leads to formation of exuberant granulation tissue (EGT) that retards healing due to protracted inflammation, aberrant vascularisation and delayed epithelialisation. EGT is not observed if wounds are left undressed or when wounds are on the body. A previous study showed that short-term administration of proteins derived from orf virus dampened inflammation and promoted epithelialisation of open wounds in horses. Here, we investigated the impact of orf virus interleukin-10 and vascular endothelial growth factor-E on the development and resolution of EGT. Excisional wounds were created on the forelimb of four horses, and bandages were maintained until full healing to induce EGT formation. Matching body wounds were created to ensure EGT was limited to the limb, and to differentiate the effects of the viral proteins on normal healing and on EGT formation. Viral proteins or the hydrogel vehicle control were administered topically to site-matched wounds at day 1, with repeat administration at day 8. Wound healing and EGT formation were monitored macroscopically. Wound margin samples were harvested at 2, 7 and 14 days, and at full healing, with histology used to observe epithelialisation, immunofluorescence used to detect inflammatory cells, angiogenesis and cell death, and qPCR to measure expression of genes regulating inflammation and angiogenesis. Limb wounds developed EGT, and exhibited slower healing than body wounds. Viral protein treatment did not accelerate healing at either location nor limit EGT formation in limb wounds. Treatment of limb wounds did however increase epithelialisation and angiogenesis, without dampening inflammatory cell infiltration or gene expression. The healed wounds also had less occlusion and death of blood vessels and fewer epidermal rete ridges following viral protein treatment. These findings indicate that the viral protein treatment does not suppress wound inflammation or EGT formation, but does promote vascular and epidermal repair and EGT resolution.


Assuntos
Membro Posterior , Cavalos , Hidrogéis/farmacologia , Interleucina-10/farmacologia , Proteínas Virais/farmacologia , Cicatrização/efeitos dos fármacos , Ferimentos e Lesões , Animais , Membro Posterior/metabolismo , Membro Posterior/patologia , Ferimentos e Lesões/tratamento farmacológico , Ferimentos e Lesões/metabolismo , Ferimentos e Lesões/patologia
13.
Virology ; 517: 199-207, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29502802

RESUMO

Monocytes are considered refractory to porcine reproductive and respiratory syndrome virus type 1 (PRRSV-1) infection. However, monocytes are only short-lived in blood, being able to differentiate into macrophages and dendritic cells (DC). It was therefore merited to revisit PRRSV-1 interaction with monocytes, particularly those treated with cytokines influencing monocyte biology. Thus, several factors were screened, particularly those modulating monocyte differentiation and expression of putative PRRSV-1 receptors (CD169 and CD163). M-CSF, known to stimulate macrophage differentiation, did not increase their susceptibility to PRRSV-1. Nor did GM-CSF or IL-4, known drivers for monocyte-derived DC (MoDC) differentiation. In contrast, monocyte treatment with IL-10 or the corticosteroid, dexamethasone, known to be potent suppressors of monocyte differentiation, was correlated with increased susceptibility to PRRSV-1 infection. While this effect was strongly correlated to CD163 and CD169 expression, our data suggest that receptor expression is not the only factor driving successful infection of PPRSV-1 in monocytes.


Assuntos
Dexametasona/farmacologia , Interleucina-10/farmacologia , Monócitos/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Animais , Antígenos CD/genética , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/genética , Antígenos de Diferenciação Mielomonocítica/metabolismo , Células Cultivadas , Meios de Cultura , Monócitos/efeitos dos fármacos , Vírus da Síndrome Respiratória e Reprodutiva Suína/classificação , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Lectina 1 Semelhante a Ig de Ligação ao Ácido Siálico/genética , Lectina 1 Semelhante a Ig de Ligação ao Ácido Siálico/metabolismo , Suínos , Cultura de Vírus , Replicação Viral
14.
Inflammation ; 41(3): 751-759, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29427162

RESUMO

Fibroblast growth factor 21 (FGF-21) has been previously judged as a major metabolic regulator. In this paper, we show that FGF-21 has a potential role in anti-inflammation and immunoregulation. In vivo, treatment with exogenous FGF-21 can alleviate LPS-induced inflammation. In vitro, FGF-21 inhibited LPS-induced IL-1ß expression in THP-1 cells. Furthermore, besides the NF-κB pathway, the mechanism of action of FGF-21 was observed to involve the elevation of IL-10 in the ERK1/2 pathway. This study clearly indicates that FGF21 can be used as an attractive target for the management of inflammatory disorders. This piece of research indicates that FGF-21 could have much value in the management of inflammatory disorders.


Assuntos
Fatores de Crescimento de Fibroblastos/farmacologia , Inflamação/tratamento farmacológico , Interleucina-10/metabolismo , Linhagem Celular , Fatores de Crescimento de Fibroblastos/fisiologia , Humanos , Inflamação/induzido quimicamente , Interleucina-10/farmacologia , Interleucina-1beta/antagonistas & inibidores , Interleucina-1beta/metabolismo , Lipopolissacarídeos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos
15.
BMC Infect Dis ; 18(1): 80, 2018 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-29439673

RESUMO

BACKGROUND: Natural killer (NK) cells play cytotoxic roles by targeting tumor cells or virus infected cells, they also play regulatory roles by secreting cytokines and chemokines. Transforming growth factor (TGF)-ß and interleukin (IL)-10 are important immunosuppressive cytokines potentially related to the immune dysregulation that occurs in the infection of human immunodeficiency virus (HIV). NK cells are an important source of TGF-ß and a main early producer of IL-10 in response to viral infection. Here, we evaluated the percentages of IL-10+ and TGF-ß+ NK cells in HIV-infected patients relative to healthy controls (HCs). METHODS: Study participants (n = 63) included 31 antiretroviral treatment (ART)-naïve HIV-infected patients, 17 ART-treated HIV-infected patients, and 15 HIV-negative HCs. Expression of IL-10 or TGF-ß in NK cells was examined by flow cytometry, and the influences of recombinant IL-10 (rIL-10) or recombinant TGF-ß (rTGF-ß) on NK cell function were investigated in vitro. RESULTS: Compared with HCs, ART-naïve HIV-infected patients had increased percentages of IL-10+ (2.0% vs. 0.4%, p = 0.015) and TGF-ß+ (4.5% vs. 2.1%, p = 0.022) NK cells, and ART-treated patients also had a higher percentage of IL-10+ NK cells (2.5% vs. 0.4%, p = 0.002). The percentages of IL-10+ and TGF-ß+ NK cells were positively correlated (r = 0.388; p = 0.010). The results of in vitro experiments demonstrated that rIL-10 and rTGF-ß inhibited NK cell CD107a expression (p = 0.037 and p = 0.024, respectively), IFN-γ secretion (p = 0.006, p = 0.016, respectively), and granzyme B release after stimulation (p = 0.014, p = 0.040, respectively). CONCLUSIONS: Our data suggest that the percentages of IL-10+ or TGF-ß+ NK cells are increased in HIV-infected patients, and that rIL-10 and/or rTGF-ß can inhibit NK cell functions in vitro, providing a potential therapeutic target for strategies aimed at combating HIV infection.


Assuntos
Infecções por HIV/patologia , Interleucina-10/metabolismo , Células Matadoras Naturais/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Adulto , Antirretrovirais/uso terapêutico , Contagem de Linfócito CD4 , Linfócitos T CD4-Positivos/citologia , Estudos de Casos e Controles , Células Cultivadas , Granzimas/metabolismo , Infecções por HIV/tratamento farmacológico , Infecções por HIV/imunologia , Humanos , Interferon gama/metabolismo , Interleucina-10/genética , Interleucina-10/farmacologia , Células Matadoras Naturais/citologia , Células Matadoras Naturais/efeitos dos fármacos , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Proteína 1 de Membrana Associada ao Lisossomo/metabolismo , Masculino , Perforina/metabolismo , RNA Viral/sangue , Proteínas Recombinantes/farmacologia , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/farmacologia , Adulto Jovem
16.
Exerc Immunol Rev ; 24: 36-44, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29461970

RESUMO

Acute respiratory distress syndrome (ARDS) is defined as hypoxemic respiratory failure with intense pulmonary inflammation, involving hyperactivation of endothelial cells and neutrophils. Given the anti-inflammatory effects of aerobic exercise (AE), this study investigated whether AE performed daily for 5 weeks would inhibit extra-pulmonary LPS-induced ARDS. C57Bl/6 mice were distributed into Control, Exercise, LPS and Exercise+LPS groups. AE was performed on a treadmill for 5x/week for four weeks before LPS administration. 24hours after the final AE physical test, animals received 100ug of LPS intra-peritoneally. In addition, whole blood cell culture, neutrophils and human endothelial cells were preincubated with IL-10, an anti-inflammatory cytokine induced by exercise. AE reduced total protein levels (p<0.01) and neutrophil accumulation in bronchoalveolar lavage (BAL) (p<0.01) and lung parenchyma (p<0.01). AE reduced BAL inflammatory cytokines IL-1ß, IL-6 and GM-CSF (p<0.001), CXCL1/KC, IL-17, TNF-alpha and IGF-1 (p<0.01). Systemically, AE reduced IL-1ß, IL-6 and IFN-gamma (p<0.001), CXCL1/KC (p<0.01) and TNF-alpha (p<0.05). AE increased IL-10 levels in serum (p<0.001) and BAL (p<0.001). Furthermore, AE increased superoxide dismutase SOD (p<0.01) and decreased superoxide anion accumulation in the lungs (p<0.01). Lastly, pre-incubation with IL-10 significantly reduced LPS-induced activation of whole blood cells, neutrophils and HUVECs, as observed by reduced production of IL-1ß, IL-6, IL-8 and TNF-alpha. Our data suggest that AE inhibited LPS-induced lung inflammation by attenuating inflammatory cytokines and oxidative stress markers in mice and human cell culture via enhanced IL-10 production.


Assuntos
Interleucina-10/imunologia , Estresse Oxidativo , Condicionamento Físico Animal , Pneumonia/imunologia , Síndrome do Desconforto Respiratório do Adulto/imunologia , Lesão Pulmonar Aguda , Animais , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Citocinas/sangue , Citocinas/imunologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Interleucina-10/farmacologia , Lipopolissacarídeos , Pulmão/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neutrófilos/imunologia , Pneumonia/induzido quimicamente , Síndrome do Desconforto Respiratório do Adulto/induzido quimicamente
17.
Mol Med Rep ; 17(4): 5700-5707, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29436649

RESUMO

Liver fibrosis is characterized by the excessive deposition of extracellular matrix (ECM) components, and activated hepatic stellate cells (HSCs) are a primary source of ECM. Several studies have revealed that the induction of HSC senescence may reduce liver fibrosis. The effect of interleukin­10 (IL­10) on the senescence of activated HSCs is not fully understood. Therefore, the present study examined its effects and potential mechanisms in activated primary rat HSCs. Collagenase perfusion and density gradient centrifugation methods were used to isolate rat HSCs. HSCs were identified by autofluorescence, Oil Red O staining and immunocytochemical analysis. Activated HSCs were treated with 0, 10, 20 or 40 ng/ml IL­10 for 24 h. Senescence­associated ß­galactosidase (SA­ß­Gal) staining, flow cytometry analysis and a cell counting kit­8 assay were performed to detect the senescence, apoptosis and viability of rat HSCs, respectively. Reverse transcription­quantitative polymerase chain reaction, western blot analysis and enzyme linked immunosorbent assays were used to detect the expression of senescence­associated proteins and cytokines. Freshly isolated rat HSCs exhibited a striking blue­green autofluorescence and HSC retinoid droplets were stained bright red by Oil Red O. Immunocytochemical analysis demonstrated the cytoplasmic expression of HSC markers desmin and α­smooth muscle actin. The number of SA­ß­Gal positive HSCs, the apoptotic rate and the expression levels of p53, p21 and tumor necrosis factor­α were significantly increased following IL­10 treatment. HSC viability and IL­6 and IL­8 expression levels were significantly decreased compared with the control group. In summary, primary rat HSCs were successfully isolated and IL­10 was demonstrated to promote the senescence of activated primary rat HSCs through the upregulation of p53 and p21 expression.


Assuntos
Senescência Celular/efeitos dos fármacos , Células Estreladas do Fígado/efeitos dos fármacos , Células Estreladas do Fígado/metabolismo , Interleucina-10/farmacologia , Proteínas Proto-Oncogênicas p21(ras)/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Animais , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Citocinas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Mediadores da Inflamação , Masculino , Proteínas Proto-Oncogênicas p21(ras)/genética , Ratos , Transdução de Sinais/efeitos dos fármacos , Proteína Supressora de Tumor p53/genética
18.
PLoS One ; 13(1): e0181912, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29357362

RESUMO

N. meningitidis induces extensive gene expression changes in human monocytes, suggesting that complex networks of signaling pathways are activated during meningococcal sepsis. These effects are modulated by the anti-inflammatory cytokine interleukin-10 (IL-10). To further study changes in signal transduction suggested by mRNA data, we used kinase substrate arrays to identify composite kinase activities induced by lysates from a primary human monocyte model system. Cell lysates were prepared from monocytes treated with the following experimental conditions: 106 N. meningitidis/mL, 25 ng/mL IL-10, 106 N. meningitidis/mL in combination with 25 ng/mL IL-10, and vehicle. Lysates were subjected to kinase activity profiling with Tyrosine Kinase PamChip® arrays containing 144 kinase peptide substrates. In our experimental model, we were not able to detect a statistically significant large-scale change in ex vivo array peptide phosphorylation by lysates from monocytes treated for 15 minutes. Targets of the IL-10 anti-inflammatory response were not identified. A profound inhibition of array peptide phosphorylation by monocytes treated for 60 minutes was identified, suggesting low activity of a large number of kinases associated with different signaling pathways and immune cell functions, including STAT3 activity, Nf-κB and VEGF signaling, and PTEN signaling activity. The peptide representing ZBTB16, which was reduced in phosphorylation by lysates from all three experimental conditions, was in Ingenuity Pathway Analysis identified to be linked to reduced cytokine release and mRNA levels of tumor necrosis factor (TNF), IL-6, and CXCL10. Further studies should investigate changes in tyrosine kinase-mediated signal transduction in human immune cells, in order to evaluate the potential clinical application of kinome profiling in the study of systemic inflammatory responses to pathogens.


Assuntos
Monócitos/enzimologia , Neisseria meningitidis/fisiologia , Proteínas Tirosina Quinases/metabolismo , Citocinas/metabolismo , Expressão Gênica , Humanos , Técnicas In Vitro , Mediadores da Inflamação/metabolismo , Interleucina-10/farmacologia , Monócitos/efeitos dos fármacos , Monócitos/microbiologia , Fosforilação , RNA Mensageiro/genética , Proteínas Recombinantes/farmacologia
19.
Mol Immunol ; 93: 206-215, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29207327

RESUMO

In response to environmental stimuli such as granulocyte-macrophage or macrophage colony stimulating factor (GM-CSF/M-CSF), macrophages (MΦ) can acquire distinct functional phenotypes that control inflammatory processes on the one hand and contribute to a broad spectrum of pathologies on the other. Potential intervention strategies will require an understanding of the signalling processes that are associated with macrophage polarization. In the present study, we show that M-MΦ produce more IFN-ß and IL-10 and a lot less TNF-α than do GM-MΦ in response to LPS. To define the molecular mechanisms that underlie the biosynthesis of TNF-α we carried out a detailed investigation of the LPS-induced activation of the canonical and non-canonical myeloid differentiation primary response 88 (MyD88)-dependent signal transduction pathways as well as the TIR-domain-containing adapter-inducing interferon-ß (TRIF)-dependent pathway. Our results show that all three pathways are activated in both cell types and that the activation is more pronounced in M-MΦ. While IL-10 was found to interfere with TNF-α production in M-MΦ, we exclude a decisive role for IFN-ß in this respect. Furthermore, we demonstrate that TNF-α mRNA is markedly destabilized in M-MΦ and that expression of the mRNA destabilizing protein tristetraprolin is greatly enhanced in these cells. Collectively, our study suggests that differential effects of LPS on TNF-α mRNA turnover and on signal transduction pathways influence the amount of TNF-α finally produced by GM-MΦ and M-MΦ.


Assuntos
Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Fator de Necrose Tumoral alfa/biossíntese , Proteínas Adaptadoras de Transporte Vesicular/fisiologia , Células Cultivadas , Regulação da Expressão Gênica , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Humanos , Interferon beta/farmacologia , Interleucina-10/farmacologia , Ativação de Macrófagos/efeitos dos fármacos , Fator Estimulador de Colônias de Macrófagos/farmacologia , Macrófagos/metabolismo , Fator 88 de Diferenciação Mieloide/fisiologia , Estabilidade de RNA , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Tristetraprolina/metabolismo , Fator de Necrose Tumoral alfa/genética
20.
Neuroreport ; 29(2): 84-91, 2018 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-29112674

RESUMO

The goal of this study was to evaluate the effects of anti-inflammatory cytokine, interleukin-10 (IL-10), and calpain inhibitor, PD150606, on the expression of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor subunits in rat hippocampal slices exposed to repeated brief hypoxic episodes. We studied both individual and combinatory effects of PD150606 and IL-10 on the expression of AMPA receptor subunits under hypoxic conditions for GluA1 and GluA2 as well as their phosphorylated forms - pSer831-GluA1 and pSer880-GluA2. Additionally, we studied whether brief hypoxic episodes and IL-10 may affect mRNA expression of transcriptional factors such as hypoxia-inducible factor-1α and nuclear factor κB (NF-κB). Western blotting analysis of hippocampal slice homogenates revealed that IL-10 and PD150606, both individually and in combination, ameliorate hypoxia-induced decrease in the expression of GluA1 and pSer831-GluA1, with different level of efficiency measured at 10, 50, and 90 min after hypoxia induction. Interestingly, brief hypoxic episodes did not induce any changes in the expression of GluA2 and pSer880-GluA2 subunits, whereas PD150606 showed biphasic effect, decreasing the expression of GluA2 and pSer880-GluA2 at 10 min and potentiating it at 90 min after hypoxia induction. IL-10 alone did not show any effect but was able to reverse PD150606 action on the expression of pSer880-GluA2 at 10 min and further potentiated it for GluA2 at 90 min after hypoxia. Finally, PCR analysis revealed that modulation of GluA1 and GluA2 expressions by hypoxia, and IL-10 was not associated with changes in the expression of hypoxia-inducible factor-1α and nuclear factor-κB (NF-κB) transcriptional factors.


Assuntos
Acrilatos/farmacologia , Fármacos do Sistema Nervoso Central/farmacologia , Hipóxia/tratamento farmacológico , Interleucina-10/farmacologia , Receptores de AMPA/metabolismo , Animais , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/fisiologia , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Hipocampo/patologia , Hipóxia/metabolismo , Hipóxia/patologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Imuno-Histoquímica , Masculino , NF-kappa B/metabolismo , RNA Mensageiro/metabolismo , Ratos Wistar , Receptores de AMPA/genética , Fatores de Tempo , Técnicas de Cultura de Tecidos
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