RESUMO
BACKGROUND: Rutaecarpine (Rut) is a plant alkaloid abundant in Euodia ruticarpa which is a Chinese herbal medicine used for treating various cancers. However, the Rut administration effect on prostate cancer in vivo remains unclear. AIM: In the present study we established an allogenic TRAMP-C1 prostate cancer mouse model to evaluate the Rut administration effect and mechanism in vivo. METHODS: To unravel the Rut administration effect on prostate cancer in vivo, C57BL/6J male mice (8 weeks old) were randomly grouped (n = 9), subcutaneously loaded with TRAMP-C1 prostate cancer cells and immediately given daily by gavage with Rut dissolved in soybean oil at 7 mg (low dose), 35 mg (medium dose), and 70 mg/kg b.w./day (high dose) for successive 39 days. RESULTS: Rut administration significantly and dose-dependently reduced both tumor volume and solid prostate cancer weight in allogenic TRAMP-C1 male mice. Rut administration markedly increased (TNF-α+IFN-γ) (Th1-)/IL-10 (Th2-) cytokine secretion ratios by splenocytes and TNF-α (M1-)/IL-10 (M2-) cytokine secretion ratios by macrophages as compared to those of dietary control group, suggesting that Rut administration in vivo regulates the immune balance toward Th1- and M1-polarized characteristics. Decreased CD19+, CD4+ and CD8+ lymphocytes in the peripheral blood of allogenic TRAMP-C1 mice were significantly elevated by Rut administration. Tumor weights positively correlated with TNF-α secretions by splenocytes, suggesting that there is a tumor cachexia in the tumor-bearing mice. Tumor weights negatively correlated with IgG (Th1-antibody) levels in the sera, suggesting that Th1-polarized immune balance may inhibit prostate cancer cell growth. CONCLUSIONS: Our results evidenced that Rut administration suppresses prostate cancer cell growth in mice subcutaneously loaded with TRAMP-C1 cells and correlated the anti-cancer effects with Th1-polarized immune balance in vivo.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Alcaloides Indólicos/farmacologia , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/genética , Quinazolinas/farmacologia , Animais , Antígenos CD/imunologia , Peso Corporal , Caquexia/patologia , Linhagem Celular Tumoral , Transformação Celular Neoplásica , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Contagem de Linfócitos , Macrófagos Peritoneais/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Tamanho do Órgão , Baço/citologia , Baço/metabolismo , Equilíbrio Th1-Th2 , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Wheat allergy is a pathological event involving immunocompetent cells against ingested wheat allergen and is clearly associated with transdermal sensitization. However, the molecular mechanisms involved in the disease etiology are not completely understood. A complex cellular and tissue network linking to food allergy makes it difficult to understand the molecular mechanism of allergenicity. Animal models are valuable tools to deduce basic principles of human disease without invasive intervention trials. A mouse model of wheat allergy has provided insights into effects of skin exposure to wheat protein; it is a plausible route of human sensitization for wheat anaphylaxis. Further investigation of this model will capture the essential occurrence and flow of events, bringing useful clues to develop effective treatment and control strategies against wheat allergy. Here, we describe a method for analyzing the expression of cell surface molecules in single cells isolated from lymphoid tissue with flow cytometry. Sensitization by wheat extracts significantly increases antigen-specific T cells in the spleen. Collecting information regarding the contribution of immune cells to allergic sensitization in the development of wheat allergy would be useful in preventing and treating food allergies.
Assuntos
Modelos Animais de Doenças , Imunofenotipagem/métodos , Linfócitos/efeitos dos fármacos , Extratos Vegetais/imunologia , Triticum/imunologia , Hipersensibilidade a Trigo/imunologia , Administração Cutânea , Animais , Antígenos CD/genética , Antígenos CD/imunologia , Biomarcadores/metabolismo , Feminino , Farinha/análise , Citometria de Fluxo , Expressão Gênica , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Interferon gama/genética , Interferon gama/imunologia , Linfonodos/citologia , Linfonodos/efeitos dos fármacos , Linfonodos/imunologia , Linfócitos/citologia , Linfócitos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Extratos Vegetais/administração & dosagem , Análise de Célula Única , Baço/citologia , Baço/efeitos dos fármacos , Baço/imunologia , Adesivo Transdérmico , Triticum/química , Hipersensibilidade a Trigo/sangue , Hipersensibilidade a Trigo/genética , Hipersensibilidade a Trigo/patologiaRESUMO
Human mesenchymal stromal/stem cells (hMSCs) show great promise in cell therapy due to their immunomodulatory properties. The overall immunomodulatory response of hMSCs resembles the resolution of inflammation, in which lipid mediators and regulatory macrophages (Mregs) play key roles. We investigated the effect of hMSC cell-cell contact and secretome on macrophages polarized and activated toward Mreg phenotype. Moreover, we studied the effect of supplemented polyunsaturated fatty acids (PUFAs): docosahexaenoic acid (DHA) and arachidonic acid, the precursors of lipid mediators, on hMSC immunomodulation. Our results show that unlike hMSC cell-cell contact, the hMSC secretome markedly increased the CD206 expression in both Mreg-polarized and Mreg-activated macrophages. Moreover, the secretome enhanced the expression of programmed death-ligand 1 on Mreg-polarized macrophages and Mer receptor tyrosine kinase on Mreg-activated macrophages. Remarkably, these changes were translated into improved Candida albicans phagocytosis activity of macrophages. Taken together, these results demonstrate that the hMSC secretome promotes the immunoregulatory and proresolving phenotype of Mregs. Intriguingly, DHA supplementation to hMSCs resulted in a more potentiated immunomodulation with increased CD163 expression and decreased gene expression of matrix metalloproteinase 2 in Mreg-polarized macrophages. These findings highlight the potential of PUFA supplementations as an easy and safe method to improve the hMSC therapeutic potential.
Assuntos
Ácido Araquidônico/farmacologia , Comunicação Celular/imunologia , Ácidos Docosa-Hexaenoicos/farmacologia , Macrófagos/imunologia , Células-Tronco Mesenquimais/imunologia , Fagocitose/efeitos dos fármacos , Antígenos CD/genética , Antígenos CD/imunologia , Antígenos de Diferenciação Mielomonocítica/genética , Antígenos de Diferenciação Mielomonocítica/imunologia , Antígeno B7-H1/genética , Antígeno B7-H1/imunologia , Candida albicans/crescimento & desenvolvimento , Candida albicans/imunologia , Comunicação Celular/efeitos dos fármacos , Polaridade Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Imunomodulação/efeitos dos fármacos , Interleucina-10/genética , Interleucina-10/imunologia , Interleucina-23/genética , Interleucina-23/imunologia , Ativação de Macrófagos/efeitos dos fármacos , Fator Estimulador de Colônias de Macrófagos/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/microbiologia , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/imunologia , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/imunologia , Células-Tronco Mesenquimais/citologia , Fenótipo , Cultura Primária de Células , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/imunologia , Receptores Imunológicos/genética , Receptores Imunológicos/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia , c-Mer Tirosina Quinase/genética , c-Mer Tirosina Quinase/imunologiaRESUMO
Hot water extract from biomass of heterotrophic mutant green alga Parachlorella kessleri HY1 (Chlorellaceae) was deproteinised, and three polysaccharidic fractions were obtained by preparative chromatography. The low-molecular fraction (1.5 × 104g mol-1) was defined mainly as branched O-2-ß-xylo-(1â3)-ß-galactofuranan where xylose is partially methylated at O-4. Two high-molecular fractions (3.05 × 105 and 9.84 × 104g mol-1) were complex polysaccharides containing α-l-rhamnan and xylogalactofuranan parts in different ratios. The polysaccharides were well soluble in hot water and, upon cooling, tended to self-segregate. Immunomodulatory activities of the obtained fractions were preliminary tested using ELISA, FACS and ImmunoSpot kits. The polysaccharides increased the TNF-α production in melanoma bearing mice with much higher intensity than in healthy mice. This was in agreement with the FACS results on T and B cells indicating their possibly secondary activation by innate immunity cells.
Assuntos
Linfócitos B/efeitos dos fármacos , Clorófitas/química , Fatores Imunológicos/farmacologia , Polissacarídeos/farmacologia , Linfócitos T/efeitos dos fármacos , Animais , Antígenos CD/genética , Antígenos CD/imunologia , Linfócitos B/imunologia , Linfócitos B/patologia , Sequência de Carboidratos , Regulação da Expressão Gênica/efeitos dos fármacos , Fatores Imunológicos/química , Fatores Imunológicos/isolamento & purificação , Interferon gama/genética , Interferon gama/imunologia , Interleucina-2/genética , Interleucina-2/imunologia , Interleucina-4/genética , Interleucina-4/imunologia , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/farmacologia , Melanoma/imunologia , Melanoma/patologia , Metilação , Camundongos , Camundongos Endogâmicos C57BL , Peso Molecular , Extratos Vegetais/química , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Cultura Primária de Células , Neoplasias Cutâneas/imunologia , Neoplasias Cutâneas/patologia , Solubilidade , Linfócitos T/imunologia , Linfócitos T/patologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia , Água , Xilose/química , Xilose/isolamento & purificaçãoRESUMO
Food allergy prevalence is increasing all over the world. Recent epidemiologic studies have shown the link between vitamin D3 insufficiency and food allergy occurrence. In this study, we investigated the effect of supplementation with cholecalciferol, a widely used form of vitamin D3, on food allergy using an experimental mouse model. In wild-type BALB/c mice which were sensitized and challenged with an experimental allergen, ovalbumin, a clinical symptom of food allergy, diarrhea, was significantly induced with the elevation of immunoglobulin E level and the increases of T helper 2 cytokine productions, such as interleukin-4, -5, and -13 (p<0.05), whereas no change in T helper 1 cytokine production, such as interferon-γ, was observed. It was also found that cell population of CD69+ CD4+ T cells was increased slightly in spleen and significantly in the mesenteric lymphnode with the diarrheal symptom (p<0.05). Treatment of cholecalciferol reduced the allergic diarrhea (p<0.05) with the decreasing tendency of CD69+ CD4+ T cells, suggesting that the cell population might be associated with the attenuating effect of cholecalciferol on diarrhea occurrence, although immunoglobulin E levels and cytokine productions were not significantly altered by the treatment of cholecalciferol. When given the mice anti-CD69 mAb treatment, significant improvement of allergic diarrhea symptom was observed (p<0.05), accompanying the decrease of CD69+ CD4+ T cells which suggested the contribution of these cells to the diarrhea symptom. Taken together, we suggest that administration of cholecalciferol might be useful to suppress symptomatic food allergy in association with the decrease of CD69+ CD4+ T cells.
Assuntos
Antígenos CD , Antígenos de Diferenciação de Linfócitos T , Linfócitos T CD4-Positivos , Colecalciferol/farmacologia , Hipersensibilidade Alimentar , Lectinas Tipo C , Animais , Antígenos CD/imunologia , Antígenos CD/metabolismo , Antígenos de Diferenciação de Linfócitos T/imunologia , Antígenos de Diferenciação de Linfócitos T/metabolismo , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Citocinas/análise , Citocinas/metabolismo , Diarreia/imunologia , Diarreia/metabolismo , Modelos Animais de Doenças , Feminino , Hipersensibilidade Alimentar/imunologia , Hipersensibilidade Alimentar/metabolismo , Lectinas Tipo C/imunologia , Lectinas Tipo C/metabolismo , Linfonodos/efeitos dos fármacos , Linfonodos/imunologia , Linfonodos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/efeitos adversos , Baço/efeitos dos fármacos , Baço/imunologia , Baço/metabolismoRESUMO
The chemokine receptor XCR1 is known to be selectively expressed by cross-presenting dendritic cells (DCs), while its ligand XCL1/lymphotactin is mainly produced by activated CD8+ T cells and natural killer cells. Recent studies have shown that XCL1-antigen fusion proteins efficiently induce CD8+ T cell responses by preferentially delivering antigens to XCR1+ DCs. However, XCL1 per se was found to be a poor adjuvant for induction of CD8+ T cell responses. XCL1 is unique because of its lack of one of the two disulfide bonds commonly conserved in all other chemokines and thus has an unstable structure with a relatively weak chemokine activity. In the present study, we generated a variant form of murine XCL1 termed mXCL1-V21C/A59C that contained a second disulfide bond to stabilize its chemokine structure. We confirmed that mXCL1-V21C/A59C had much more potent chemotactic and calcium mobilization activities than the wild type XCL1 (mXCL1-WT). Intradermal injection of mXCL1-V21C/A59C, but not that of mXCL1-WT, significantly increased the accumulation of XCR1+CD103+ DCs in the injection site, and most of the accumulated XCR1+CD103+ DCs were found to take up co-injected ovalbumin (OVA). Furthermore, recruited XCR1+CD103+ DCs efficiently migrated to the draining lymph nodes and stayed for a prolonged period of time. Consequently, mXCL1-V21C/A59C strongly induced OVA-specific CD8+ T cells. The combination of OVA and mXCL1-V21C/A59C well protected mice from E.G7-OVA tumor growth in both prophylactic and therapeutic protocols. Finally, memory CTL responses were efficiently induced in mice immunized with OVA and mXCL1-V21C/A59C. Although intradermal injection of OVA and polyinosinic-polycytidylic acid (poly(I:C)) as an adjuvant also induced CD8+ T cell responses to OVA, poly (I:C) poorly recruited XCR1+CD103+ DCs in the injection site and failed to induce significant memory CTL responses to OVA. Collectively, our findings demonstrate that a highly active form of XCL1 is a promising vaccine adjuvant for cross-presenting DCs to induce antigen-specific effector and memory CD8+ T cells.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Quimiocinas C/imunologia , Apresentação Cruzada/imunologia , Células Dendríticas/imunologia , Memória Imunológica/imunologia , Linfocinas/imunologia , Sialoglicoproteínas/imunologia , Adjuvantes Imunológicos/farmacologia , Animais , Antígenos/imunologia , Antígenos CD/imunologia , Cálcio/imunologia , Linhagem Celular , Apresentação Cruzada/efeitos dos fármacos , Células Dendríticas/efeitos dos fármacos , Memória Imunológica/efeitos dos fármacos , Cadeias alfa de Integrinas/imunologia , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Ovalbumina/imunologiaRESUMO
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, ÐÐ 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/imunologiaRESUMO
Honokiol is a biphenolic isolate extracted from the bark of the magnolia tree that has been used in traditional Chinese and Japanese medicine, and has more recently been investigated for its anti-inflammatory and antibacterial properties. Honokiol has previously been demonstrated to improve survival in sepsis models that have rapid 100% lethality. The purpose of this study was to determine the impact of Honokiol on the host response in a model of sepsis that more closely approximates human disease. Male and female C57BL/6 mice underwent cecal ligation and puncture to induce polymicrobial intra-abdominal sepsis. Mice were then randomized to receive an injection of either Honokiol (120âmg/kg/day) or vehicle and were sacrificed after 24âh for functional studies or followed 7 days for survival. Honokiol treatment after sepsis increased the frequency of CD4 T cells and increased activation of CD4 T cells as measured by the activation marker CD69. Honokiol also increased splenic dendritic cells. Honokiol simultaneously decreased frequency and number of CD8 T cells. Honokiol decreased systemic tumor necrosis factor without impacting other systemic cytokines. Honokiol did not have a detectable effect on kidney function, lung physiology, liver function, or intestinal integrity. In contrast to prior studies of Honokiol in a lethal model of sepsis, Honokiol did not alter survival at 7 days (70% mortality for Honokiol vs. 60% mortality for vehicle). Honokiol is thus effective in modulating the host immune response and inflammation following a clinically relevant model of sepsis but is not sufficient to alter survival.
Assuntos
Compostos de Bifenilo/farmacologia , Linfócitos T CD4-Positivos/imunologia , Lignanas/farmacologia , Ativação Linfocitária/efeitos dos fármacos , Sepse/tratamento farmacológico , Fator de Necrose Tumoral alfa/imunologia , Animais , Antígenos CD/imunologia , Antígenos de Diferenciação de Linfócitos T/imunologia , Linfócitos T CD4-Positivos/patologia , Linfócitos T CD8-Positivos/imunologia , Modelos Animais de Doenças , Feminino , Inflamação/tratamento farmacológico , Inflamação/imunologia , Inflamação/patologia , Lectinas Tipo C/imunologia , Contagem de Linfócitos , Masculino , Camundongos , Distribuição Aleatória , Sepse/imunologia , Sepse/patologiaRESUMO
Dendritic cells (DCs) in mesenteric lymph nodes (MLNs) induce Foxp3+ regulatory T cells to regulate immune responses to beneficial or non-harmful agents in the intestine, such as commensal bacteria and foods. Several studies in MLN DCs have revealed that the CD103+ DC subset highly induces regulatory T cells, and another study has reported that MLN DCs from programmed death ligand 1 (PD-L1) -deficient mice could not induce regulatory T cells. Hence, the present study investigated the expression of these molecules on MLN CD11c+ cells. Four distinct subsets expressing CD103 and/or PD-L1 were identified, namely CD11b+ CD103+ PD-L1High , CD11b- CD103+ PD-L1High , CD11b- CD103+ PD-L1Low and CD11b+ CD103- PD-L1Int . Among them, the CD11b- CD103+ PD-L1High DC subset highly induced Foxp3+ T cells. This subset expressed Aldh1a2 and Itgb8 genes, which are involved in retinoic acid metabolism and transforming growth factor-ß (TGF-ß) activation, respectively. Exogenous TGF-ß supplementation equalized the level of Foxp3+ T-cell induction by the four subsets whereas retinoic acid did not, which suggests that high ability to activate TGF-ß is determinant for the high Foxp3+ T-cell induction by CD11b- CD103+ PD-L1High DC subset. Finally, this subset exhibited a migratory DC phenotype and could take up and present orally administered antigens. Collectively, the MLN CD11b- CD103+ PD-L1High DC subset probably takes up luminal antigens in the intestine, migrates to MLNs, and highly induces regulatory T cells through TGF-ß activation.
Assuntos
Antígenos CD/imunologia , Antígeno B7-H1/imunologia , Antígeno CD11b/imunologia , Comunicação Celular , Células Dendríticas/imunologia , Cadeias alfa de Integrinas/imunologia , Intestinos/imunologia , Linfonodos/imunologia , Linfócitos T Reguladores/imunologia , Administração Oral , Aldeído Desidrogenase/imunologia , Aldeído Desidrogenase/metabolismo , Família Aldeído Desidrogenase 1 , Animais , Antígenos CD/metabolismo , Antígeno B7-H1/metabolismo , Antígeno CD11b/metabolismo , Comunicação Celular/efeitos dos fármacos , Movimento Celular , Células Cultivadas , Técnicas de Cocultura , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/metabolismo , Fatores de Transcrição Forkhead/imunologia , Fatores de Transcrição Forkhead/metabolismo , Imunidade nas Mucosas , Cadeias alfa de Integrinas/metabolismo , Cadeias beta de Integrinas/imunologia , Cadeias beta de Integrinas/metabolismo , Mucosa Intestinal/metabolismo , Intestinos/citologia , Intestinos/efeitos dos fármacos , Linfonodos/citologia , Linfonodos/efeitos dos fármacos , Linfonodos/metabolismo , Mesentério , Camundongos Endogâmicos BALB C , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Fenótipo , Retinal Desidrogenase , Transdução de Sinais , Linfócitos T Reguladores/efeitos dos fármacos , Linfócitos T Reguladores/metabolismo , Fator de Crescimento Transformador beta/imunologia , Fator de Crescimento Transformador beta/metabolismo , Tretinoína/farmacologiaRESUMO
Cancers are able to grow by subverting immune suppressive pathways, to prevent the malignant cells as being recognized as dangerous or foreign. This mechanism prevents the cancer from being eliminated by the immune system and allows disease to progress from a very early stage to a lethal state. Immunotherapies are newly developing interventions that modify the patient's immune system to fight cancer, by either directly stimulating rejection-type processes or blocking suppressive pathways. Extracellular adenosine generated by the ectonucleotidases CD39 and CD73 is a newly recognized "immune checkpoint mediator" that interferes with anti-tumor immune responses. In this review, we focus on CD39 and CD73 ectoenzymes and encompass aspects of the biochemistry of these molecules as well as detailing the distribution and function on immune cells. Effects of CD39 and CD73 inhibition in preclinical and clinical studies are discussed. Finally, we provide insights into potential clinical application of adenosinergic and other purinergic-targeting therapies and forecast how these might develop in combination with other anti-cancer modalities.
Assuntos
5'-Nucleotidase/metabolismo , Anticorpos Monoclonais/uso terapêutico , Antígenos CD/metabolismo , Apirase/metabolismo , Imunoterapia/métodos , Neoplasias/terapia , 5'-Nucleotidase/imunologia , Animais , Antígenos CD/imunologia , Apirase/imunologia , Ensaios Clínicos como Assunto , Terapia Combinada , Avaliação Pré-Clínica de Medicamentos , Proteínas Ligadas por GPI/imunologia , Proteínas Ligadas por GPI/metabolismo , Humanos , Neoplasias/imunologia , Evasão Tumoral , Microambiente TumoralRESUMO
Activating innate immunity by an adjuvant is required in vaccine development. The study aims to investigate adjuvant effects of aqueous extracts of Artemisia rupestris L. (AEAR) in vivo and in vitro. ICR mice were subcutaneously administered with antigen and AEAR at various doses to evaluate their immune responses of antibodies, dendritic cells (DCs), regulatory T cells (Treg), splenic lymphocyte, and cytokine. The evaluation results showed that AEAR could largely increase titers of antigen-specific antibodies (IgG, IgG1, and IgG2a) and T cell proliferation. AEAR also increased expression of IFN-γ in CD8+T cells as well as IL-4 and INF-γ expression in CD4+T cells. Expression levels of MHC-II, CD40, CD80, and CD86 on DCs were significantly elevated, whereas the Treg frequency was significantly decreased. AEAR (200µg) showed remarkable adjuvant activity. Furthermore, AEAR enhanced MHC-II, CD40, CD80, and CD86 expression as well as the yields of TNF-α and IL-12 on DCs through toll-like receptor4 (TLR4) in vitro. Those results indicated that AEAR could serve as an efficacious immune stimulator for vaccines because it significantly enhanced specific immune responses by promoting DCs maturation and reduced Treg through TLR4 signaling pathway.
Assuntos
Adjuvantes Imunológicos/farmacologia , Artemisia/química , Ovalbumina/imunologia , Transdução de Sinais/efeitos dos fármacos , Linfócitos T Reguladores/efeitos dos fármacos , Receptor 4 Toll-Like/imunologia , Animais , Antígenos CD/genética , Antígenos CD/imunologia , Células Dendríticas/citologia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Feminino , Regulação da Expressão Gênica , Antígenos de Histocompatibilidade Classe II/genética , Antígenos de Histocompatibilidade Classe II/imunologia , Imunoglobulina G/biossíntese , Interferon gama/genética , Interferon gama/imunologia , Interleucina-12/genética , Interleucina-12/imunologia , Interleucina-4/genética , Interleucina-4/imunologia , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos ICR , Ovalbumina/administração & dosagem , Extratos Vegetais/química , Baço/citologia , Baço/efeitos dos fármacos , Baço/imunologia , Linfócitos T Reguladores/citologia , Linfócitos T Reguladores/imunologia , Receptor 4 Toll-Like/genética , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
BACKGROUND: Semaphorin 7A (Sema7A) is expressed by several different classes of lymphoid and myeloid cells and is a potent immunomodulator. We examined the role of Sema7A in modulating cellular immune responses and to provide experimental data validating the therapeutic potential of Sema7A in rheumatoid arthritis (RA). METHODS: Soluble Sema7A (sSema7A) levels in the serum and synovial fluid from patients with RA or osteoarthritis, as well as cytokine secretions, were analyzed with an enzyme-linked immunosorbent assay. The cell surface levels and transcripts of Sema7A were evaluated in T cells and monocytes from patients with RA. The effect of Sema7A on the functions of primary T cells isolated from the peripheral blood of healthy donors was observed. Detection of the activation of the signal mediator focal adhesion kinase was performed by Western blotting. Shedding of sSema7A was evaluated in monocytes. The introduction of anti-Sema7A antibody to mice with collagen-induced arthritis (CIA) was observed in vivo. RESULTS: Upregulation of sSema7A levels in both the serum and synovial fluid of patients with RA was correlated with disease activity markers. sSema7A markedly increased Th1/Th17 cytokine secretion and induced evident upregulation of T-bet and retinoic acid receptor-related orphan nuclear receptor γt levels in T cells. Cell surface Sema7A was cleaved by a disintegrin and metalloprotease 17 (ADAM17) in monocytes. Interleukin-6 and tumor necrosis factor-α stimulated ADAM17 secretion in synovial macrophages. Blocking of ß1-integrin abrogated the Sema7A-mediated cytokine secretion. Treatment with an anti-Sema7A antibody significantly attenuated CIA. CONCLUSIONS: These findings indicate that Sema7A as a potent activator of T cells and monocytes in the immune response contributes to the inflammation and progression of RA, suggesting its therapeutic potential in the treatment of RA.
Assuntos
Antígenos CD/imunologia , Antígenos CD/metabolismo , Artrite Reumatoide/imunologia , Artrite Reumatoide/metabolismo , Semaforinas/imunologia , Semaforinas/metabolismo , Animais , Antígenos CD/análise , Artrite Experimental/imunologia , Artrite Experimental/metabolismo , Western Blotting , Separação Celular , Ensaio de Imunoadsorção Enzimática , Proteínas Ligadas por GPI/análise , Proteínas Ligadas por GPI/imunologia , Proteínas Ligadas por GPI/metabolismo , Técnicas de Silenciamento de Genes , Humanos , Camundongos , Camundongos Endogâmicos DBA , Monócitos/imunologia , Monócitos/metabolismo , Reação em Cadeia da Polimerase , Semaforinas/análise , Linfócitos T/imunologia , Linfócitos T/metabolismoRESUMO
OBJECTIVE: To explore the therapeutic effect and possible mechanism of Huangqi glycoprotein (HQGP) on the development of experimental autoimmune encephalomyelitis (EAE). METHODS: Female C57BL/6 mice were immunized subcutaneously with myelin oligodendrocyte glycoprotein peptide-35-55 (MOG35-55) and divided into HQGP-treated group and EAE control group. Clinical score and body mass were recorded every other day. Inflammatory cell infiltrations of spinal cord were observed by HE and immunofluorescence staining. The cell viability of splenic mononuclear cells (MNCs) was detected by MTT assay. The release of NO was measured by Griess method. The levels of TNF-α, IL-6 and IFN-γ were determined by ELISA. The subtypes of CD4(+)T cells were analyzed by flow cytometry. RESULTS: HQGP treatment delayed the onset of EAE, attenuated the clinical symptoms and inhibited CD68(+) macrophage infiltration into the central nervous system. HQGP inhibited the viability of splenic MNCs, downregulated the secretion of NO, TNF-α, IL-6, and increased the secretion of IFN-γ. In addition, HQGP treatment effectively increased the numbers of CD4(+)CD25(+) T cells, CD4(+)IL-10(+) T cells and CD4(+)IFN-γ(+) T cells. CONCLUSION: HQGP relieve the inflammation of EAE via reducing the number of T cell subsets and inhibiting the secretion of inflammatory cytokines.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Encefalomielite Autoimune Experimental/prevenção & controle , Glicoproteínas/farmacologia , Fatores Imunológicos/farmacologia , Animais , Antígenos CD/imunologia , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/imunologia , Antígenos de Diferenciação Mielomonocítica/metabolismo , Astragalus propinquus/química , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Citocinas/imunologia , Citocinas/metabolismo , Encefalomielite Autoimune Experimental/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Imunofluorescência , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos Endogâmicos C57BL , Glicoproteína Mielina-Oligodendrócito/imunologia , Óxido Nítrico/imunologia , Óxido Nítrico/metabolismo , Medula Espinal/efeitos dos fármacos , Medula Espinal/imunologia , Medula Espinal/metabolismo , Baço/efeitos dos fármacos , Baço/imunologia , Baço/metabolismo , Subpopulações de Linfócitos T/efeitos dos fármacos , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismoRESUMO
Increased risk of allergy during early life indicates deficient immune regulation in this period of life. To date, the cause for inefficient neonatal immune regulation has never been elucidated. We aimed to define the ontogeny of oral tolerance and to identify necessary conditions specific for this stage of life. Ovalbumin (OVA) was administered orally to mice through breast milk and efficiency of systemic tolerance to OVA was assessed in adulthood using a model of allergic airway inflammation. Oral tolerance induction was fully efficient starting third week of life. Inefficiency in neonates was a consequence of abnormal antigen transfer across the gut barrier and retinaldehyde dehydrogenase expression by mesenteric lymph node CD103(+) neonatal dendritic cells, resulting in inefficient T-cell activation. Neonates' serum retinol levels were three times lower than in adult mice, and vitamin A supplementation was sufficient to rescue neonatal defects and allow tolerance induction from birth. The establishment of oral tolerance required the differentiation of Th1 lymphocytes in both vitamin A-supplemented neonates and 3-week-old unsupplemented mice. This knowledge should guide the design of interventions for allergy prevention that are adapted to the neonatal stage of life such as vitamin A supplementation.
Assuntos
Tolerância Imunológica/efeitos dos fármacos , Ovalbumina/farmacologia , Células Th1/imunologia , Deficiência de Vitamina A/prevenção & controle , Vitamina A/administração & dosagem , Administração Oral , Animais , Animais Recém-Nascidos , Animais Lactentes , Antígenos CD/genética , Antígenos CD/imunologia , Células Dendríticas/citologia , Células Dendríticas/imunologia , Expressão Gênica , Cadeias alfa de Integrinas/genética , Cadeias alfa de Integrinas/imunologia , Linfonodos/citologia , Linfonodos/imunologia , Ativação Linfocitária , Mesentério/citologia , Mesentério/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Linfócitos T Reguladores/citologia , Linfócitos T Reguladores/imunologia , Células Th1/citologia , Vitamina A/imunologia , Vitamina A/metabolismo , Deficiência de Vitamina A/imunologia , Deficiência de Vitamina A/fisiopatologiaRESUMO
In general, detection of peritoneal carcinomatosis (PC) occurs at the late stage when there is no treatment option. In the present study, we designed novel drug delivery systems that are functionalized with anti-CD133 antibodies. The C1, C2 and C3 complexes with cisplatin were introduced into nanotubes, either physically or chemically. The complexes were reacted with anti-CD133 antibody to form the labeled product of A0-o-CX-chem-CD133. Cytotoxicity screening of all the complexes was performed on CHO cells. Data showed that both C2 and C3 Pt-complexes are more cytotoxic than C1. Flow-cytometry analysis showed that nanotubes conjugated to CD133 antibody have the ability to target cells expressing the CD133 antigen which is responsible for the emergence of resistance to chemotherapy and disease recurrence. The shortest survival rate was observed in the control mice group (K3) where no hyperthermic intraperitoneal chemotherapy procedures were used. On the other hand, the longest median survival rate was observed in the group treated with A0-o-C1-chem-CD133. In summary, we designed a novel drug delivery system based on carbon nanotubes loaded with Pt-prodrugs and functionalized with anti-CD133 antibodies. Our data demonstrates the effectiveness of the new drug delivery system and provides a novel therapeutic modality in the treatment of melanoma.
Assuntos
Cisplatino/administração & dosagem , Cisplatino/química , Sistemas de Liberação de Medicamentos/métodos , Hipertermia Induzida/métodos , Nanotubos de Carbono/química , Neoplasias Peritoneais/terapia , Antígeno AC133 , Animais , Anticorpos/administração & dosagem , Anticorpos/química , Anticorpos/imunologia , Antígenos CD/química , Antígenos CD/imunologia , Terapia Combinada , Modelos Animais de Doenças , Glicoproteínas/química , Glicoproteínas/imunologia , Imunotoxinas/administração & dosagem , Imunotoxinas/química , Imunotoxinas/imunologia , Injeções Intraperitoneais , Melanoma Experimental/tratamento farmacológico , Melanoma Experimental/terapia , Camundongos , Camundongos Endogâmicos C57BL , Peptídeos/química , Peptídeos/imunologia , Neoplasias Peritoneais/tratamento farmacológico , Taxa de SobrevidaRESUMO
In recent years, several studies have shown that Rhodiola rosea can enhance cellular immunity and humoral immune function in mice, and thus, it has become a research hotspot. However, its underlying mechanism of action has remained elusive. The present study investigated whether Rhodiola rosea was able to downregulate the expression of tumor necrosis factor-α-inducible protein 8-like 2 (TIPE2), thereby inhibiting the expression of apoptotic genes, attenuating T-lymphocyte apoptosis and improving immunity in septic mice. A mouse model of caecal ligation and puncture (CLP)-induced sepsis was established, and animals in the treatment group were pre-treated with an intraperitoneal injection of Rhodiola rosea extract, while animals in the control group and sham-operated group were injected with an equivalent amount of normal saline. TIPE2, B-cell lymphoma 2 (Bcl-2), Fas and Fas ligand (FasL) mRNA and protein levels in thymic T cells were determined using reverse transcription quantitative polymerase chain reaction and western blot analysis, respectively. Furthermore, the thymus T-lymphocyte apoptosis rate, thymus T-lymphocyte count and thymus T-lymphocyte sub-sets were assessed using flow cytometry. Levels of T-helper cell type 1 (Th1) cytokines [Interleukin (IL)-2, IL-12 and interferon (IFN)-γ] and Th2 cytokines (IL-4 and IL-10) were determined using ELISA. The results showed that, compared to that in the CLP group, the expression of TIPE2, Fas and FasL in the treatment group was significantly decreased, while the expression of Bcl-2 was increased (P<0.05). The thymus lymphocyte count in the CLP group was significantly higher compared with that in the treatment group (P<0.05). Furthermore, the apoptotic rate of thymus T-lymphocytes in the treatment group was significantly lower than that in the CLP group (P<0.05). In addition, treatment with Rhodiola rosea rescued decreased in the counts of the CD3(+) T and CD4(+) T sub-sets of thymus T lymphocytes in the CLP group (P<0.05), while not affecting the increased levels of Th2 cytokines (IL-4 and IL-10) in the CLP group compared with those in the control groups. In addition, the Th1 cytokines (IL-12, IL-2 and IFN-γ) were significantly increased (P<0.05) in the CLP group, and treatment with Rhodiola rosea led to further increases. The thymus index of septic mice treated with Rhodiola rosea as well as their survival rate were improved as compared with those in the CLP group. These findings suggested that Rhodiola rosea has protective effects against sepsis by decreasing apoptosis, increasing Th1 cytokines and enhancing the host's immunity via the regulation of TIPE2 expression.
Assuntos
Fatores Imunológicos/uso terapêutico , Peptídeos e Proteínas de Sinalização Intracelular/genética , Extratos Vegetais/uso terapêutico , Rhodiola/química , Sepse/tratamento farmacológico , Linfócitos T/efeitos dos fármacos , Timo/efeitos dos fármacos , Animais , Antígenos CD/análise , Antígenos CD/imunologia , Apoptose/efeitos dos fármacos , Citocinas/análise , Citocinas/imunologia , Regulação para Baixo/efeitos dos fármacos , Feminino , Deleção de Genes , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Peptídeos e Proteínas de Sinalização Intracelular/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Ratos , Sepse/genética , Sepse/imunologia , Sepse/patologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Linfócitos T/patologia , Timo/imunologia , Timo/metabolismo , Timo/patologiaRESUMO
The Fc receptor I for IgA (FcαRI) down-regulates humoral immune responses and modulates the risk of autoimmunity. This study aimed to investigate whether FcαRI targeting can affect progression of pristine-induced lupus nephritis. In the first experiment (early intervention), four groups of animals were evaluated: untreated FcαRI/FcRγ transgenic (Tg) mice and Tg mice administered control antibody (Ctr Fab), saline and anti-FcαRI Fab [macrophage inflammatory protein (MIP)-8a], respectively, three times a week for 29 weeks, after being injected once intraperitoneally with 0·5 ml pristane. In the second experiment, antibody injection started after the onset of nephritis and was carried out for 2 months, with similar groups as described above. MIP-8a improved proteinuria, decreased the amounts of glomerular injury markers, serum interleukin (IL)-6, IL-1 and monocyte chemoattractant protein (MCP)-1, and F4/80 macrophages in the interstitium and glomeruli, in both experiments. When MIP-8a was used as early intervention, a decrease in mouse serum anti-nuclear antibody (ANA) titres and reduced deposition of immunoglobulins in glomeruli were observed. This effect was associated with reduced serum titres of immunoglobulin (Ig)G2a but not IgG1, IgG2b and IgG3. Furthermore, pathological analysis showed lower glomerular activity index and less fibronectin in MIP-8a treated mice. This study suggests that FcαRI targeting could halt disease progression and lupus activation by selective inhibition of cytokine production, leucocyte recruitment and renal inflammation. Our findings provide a basis for the use of FcαRI as a molecular target for the treatment of lupus.
Assuntos
Anticorpos Monoclonais/farmacologia , Nefrite Lúpica/prevenção & controle , Terapia de Alvo Molecular/métodos , Receptores Fc/antagonistas & inibidores , Animais , Anticorpos Monoclonais/imunologia , Antígenos CD/genética , Antígenos CD/imunologia , Autoanticorpos/sangue , Autoanticorpos/imunologia , Citocinas/sangue , Citocinas/imunologia , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Rim/efeitos dos fármacos , Rim/patologia , Rim/ultraestrutura , Nefrite Lúpica/induzido quimicamente , Nefrite Lúpica/imunologia , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Microscopia Eletrônica , Receptores Fc/genética , Receptores Fc/imunologia , Terpenos , Fatores de TempoRESUMO
Accumulating evidences from animal studies have indicated that both endogenous and exogenous IL-27, an IL-12 family of cytokine, can increase antitumor T-cell activities and inhibit tumor growth. IL-27 can modulate Treg responses, and program effector T cells into a unique T-effector stem cell (TSEC) phenotype, which enhances T-cell survival in the tumor microenvironment. However, animal studies also suggest that IL-27 induces molecular pathways such as IL-10, PD-L1 and CD39, which may downregulate tumor-specific T-cell responses. In this review paper, we will discuss the Yin and Yang aspects of IL-27 in the induction of tumor-specific T-cell responses, and the potential impacts of these functions of IL-27 in the design of cancer immunotherapy.
Assuntos
Imunoterapia/métodos , Interleucinas/imunologia , Células Progenitoras Linfoides/imunologia , Neoplasias/terapia , Linfócitos T Reguladores/imunologia , Microambiente Tumoral/imunologia , Antígenos CD/imunologia , Apirase/imunologia , Antígeno B7-H1/imunologia , Humanos , Interleucina-10/imunologia , Células Progenitoras Linfoides/patologia , Neoplasias/imunologia , Neoplasias/patologia , Linfócitos T Reguladores/patologiaRESUMO
In this study, the new stable sulfur-containing compounds onionins A2 (1) and A3 (2) were isolated from the acetone extracts of the bulbs of Allium cepa L. and identified as the stereoisomers of onionin A1 discovered in our previous study. Their chemical structures, 3,4-dimethyl-5-(1E-propenyl)-tetrahydrothiophene-2-sulfenic acid-S-oxides, were characterized using various spectroscopic techniques. In addition, 1 and 2 together with onionin A1 were successfully isolated from the leaves of the Welsh onion, Allium fistulosum L. The onion-extracted fractions showed good potential to inhibit the polarization of M2 activated macrophages, indicating their possible ability to inhibit tumor cell proliferation.
Assuntos
Fatores Imunológicos/química , Macrófagos/efeitos dos fármacos , Cebolas/química , Ácidos Sulfênicos/química , Tiofenos/química , Antígenos CD/análise , Antígenos CD/imunologia , Antígenos de Diferenciação Mielomonocítica/análise , Antígenos de Diferenciação Mielomonocítica/imunologia , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Antineoplásicos Fitogênicos/farmacologia , Células Cultivadas , Humanos , Fatores Imunológicos/isolamento & purificação , Fatores Imunológicos/farmacologia , Macrófagos/imunologia , Raízes de Plantas/química , Receptores de Superfície Celular/análise , Receptores de Superfície Celular/imunologia , Ácidos Sulfênicos/isolamento & purificação , Ácidos Sulfênicos/farmacologia , Tiofenos/isolamento & purificação , Tiofenos/farmacologiaRESUMO
Conventional chemotherapy of ovarian cancer often fails because of initiation of drug resistance and/or side effects and trace of untouched remaining cancerous cells. This highlights an urgent need for advanced targeted therapies for effective remediation of the disease using a cytotoxic agent with immunomodulatory effects, such as shikonin (SHK). Based on preliminary experiments, we found SHK to be profoundly toxic in ovarian epithelial cancer cells (OVCAR-5 and ID8 cells) as well as in normal ovarian IOSE-398 cells, endothelial MS1 cells, and lymphocytes. To limit its cytotoxic impact solely to tumor cells within the tumor microenvironment (TME), we aimed to engineer SHK as polymeric nanoparticles (NPs) with targeting moiety toward tumor microvasculature. To this end, using single/double emulsion solvent evaporation/diffusion technique with sonication, we formulated biodegradable NPs of poly(lactic-co-glycolic acid) (PLGA) loaded with SHK. The surface of NPs was further decorated with solubilizing agent polyethylene glycol (PEG) and tumor endothelial marker 1 (TEM1)/endosialin-targeting antibody (Ab) through carbodiimide/N-hydroxysuccinimide chemistry. Having characterized the physicochemical and morphological properties of NPs, we studied their drug-release profiles using various kinetic models. The biological impact of NPs was also evaluated in tumor-associated endothelial MS1 cells, primary lymphocytes, and epithelial ovarian cancer OVCAR-5 cells. Based on particle size analysis and electron microscopy, the engineered NPs showed a smooth spherical shape with size range of 120 to 250 nm and zeta potential value of -30 to -40 mV. Drug entrapment efficiency was ~80%-90%, which was reduced to ~50%-60% upon surface decoration with PEG and Ab. The liberation of SHK from NPs showed a sustained-release profile that was best fitted with Wagner log-probability model. Fluorescence microscopy and flow cytometry analysis showed active interaction of Ab-armed NPs with TEM1-positive MS1 cells, but not with TEM1-negative MS1 cells. While exposure of the PEGylated NPs for 2 hours was not toxic to lymphocytes, long-term exposure of the Ab-armed and PEGylated NPs was significantly toxic to TEM1-positive MS1 cells and OVCAR-5 cells. Based on these findings, we propose SHK-loaded Ab-armed PEGylated PLGA NPs as a novel nanomedicine for targeted therapy of solid tumors.