RESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: The traditional Chinese medicine (TCM) XYQFT is composed of 10 herbs. According to the NHIRD, XYQFT is one of the top ten most commonly used TCM prescriptions for asthma treatment. AIM OF THE STUDY: The aim of this study was to explore whether XYQFT reduces asthma symptoms in a mouse model of chronic asthma and determine the immunomodulatory mechanism of mast cells. MATERIALS AND METHODS: BALB/c mice were intratracheally (it) stimulated with 40 µL (2.5 µg/µL) of Dermatophagoides pteronyssinus (Der p) once a week for 6 consecutive weeks and orally administered XYQFT at 1 g/kg 30 min before Der p stimulation. Airway hypersensitivity, inflammatory cells in the BALF and total IgE in the blood were assessed in mice. In addition, RBL-2H3 cells (mast cells) were stimulated with DNP-IgE, after which different concentrations of XYQFT were added for 30 min to evaluate the effect of XYQFT on the gene expression and degranulation of DNP-stimulated RBL-2H3 cells. After the compounds in XYQFT were identified using LCâMS/MS, the PBD method was used to identify the chemical components that inhibited the expression of the GM-CSF and COX-2 genes in mast cells. RESULTS: The airway hypersensitivity assay demonstrated that XYQFT significantly alleviated Der p-induced airway hypersensitivity. Moreover, cell counting and typing of bronchoalveolar lavage fluid revealed a significant reduction in Der p-induced inflammatory cell infiltration with XYQFT treatment. ELISA examination further indicated a significant decrease in Der p-induced total IgE levels in serum following XYQFT administration. In addition, XYQFT inhibited the degranulation and expression of genes (IL-3, IL-4, ALOX-5, IL-13, GM-CSF, COX-2, TNF-α, and MCP-1) in RBL-2H3 cells after DNP stimulation. The compounds timosaponin AIII and genkwanin in XYQFT were found to be key factors in the inhibition of COX-2 and GM-CSF gene expression in mast cells. CONCLUSION: By regulating mast cells, XYQFT inhibited inflammatory cell infiltration, airway hypersensitivity and specific immunity in a mouse model of asthma. In addition, XYQFT synergistically inhibited the expression of the GM-CSF and COX-2 genes in mast cells through timosaponin AIII and genkwanin.
Asunto(s)
Asma , Ciclooxigenasa 2 , Medicamentos Herbarios Chinos , Factor Estimulante de Colonias de Granulocitos y Macrófagos , Mastocitos , Animales , Masculino , Ratones , Ratas , Antiasmáticos/farmacología , Asma/tratamiento farmacológico , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Línea Celular , Ciclooxigenasa 2/metabolismo , Ciclooxigenasa 2/genética , Modelos Animales de Enfermedad , Medicamentos Herbarios Chinos/farmacología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Inmunoglobulina E/sangre , Mastocitos/efectos de los fármacos , Mastocitos/metabolismo , Ratones Endogámicos BALB CRESUMEN
Metabolic adaptations can directly influence the scope and scale of macrophage activation and polarization. Here we explore the impact of type I interferon (IFNß) on macrophage metabolism and its broader impact on cytokine signaling pathways. We find that IFNß simultaneously increased the expression of immune-responsive gene 1 and itaconate production while inhibiting isocitrate dehydrogenase activity and restricting α-ketoglutarate accumulation. IFNß also increased the flux of glutamine-derived carbon into the tricarboxylic acid cycle to boost succinate levels. Combined, we identify that IFNß controls the cellular α-ketoglutarate/succinate ratio. We show that by lowering the α-ketoglutarate/succinate ratio, IFNß potently blocks the JMJD3-IRF4-dependent pathway in GM-CSF and IL-4 activated macrophages. The suppressive effects of IFNß on JMJD3-IRF4-dependent responses, including M2 polarization and GM-CSF-induced inflammatory pain, were reversed by supplementation with α-ketoglutarate. These results reveal that IFNß modulates macrophage activation and polarization through control of the cellular α-ketoglutarate/succinate ratio.
Asunto(s)
Interferón Tipo I , Activación de Macrófagos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Ácidos Cetoglutáricos/metabolismo , Ácidos Cetoglutáricos/farmacología , Ácido SuccínicoRESUMEN
The consumption of probiotics and fermented foods has been very popular in recent decades. The primary aim of our study was to evaluate the effect of probiotics on the gut microbiota and the changes in inflammatory cytokines after an average of 6.7 weeks of probiotic administration among normal pregnant women. Thirty-two healthy pregnant women at 32 weeks of gestation were recruited and divided into two groups. The probiotic group ingested combined probiotics until after birth. The base characteristics of the probiotics and control groups showed no significant differences. The structure of the fecal microbiota at the genus level varied during the third trimester, and administration of probiotics had no influence on the composition of the fecal microbiota however, many highly abundant taxa and core microbiota at the genus level changed in the probiotic group when compared to the control group. The analysis of cytokines showed that IL-5, IL-6, TNF-α, and GM-CSF had equal levels between the baseline and control groups but were significantly increased after probiotic administration (baseline = control < probiotics). Additionally, levels of IL-1ß, IL-2, IL-12, and IFN-γ significantly increased among the three groups (baseline < control < probiotics). This result demonstrated that probiotics helped to shift the anti-inflammatory state to a pro-inflammatory state. The correlation analysis outcome suggested that the relationship between the microbiota and the cytokines was not strain-dependent. The gut microbiota varied during the third trimester. The probiotics demonstrated immunomodulation effects that helped to switch over to a pro-inflammatory immune state in the third trimester, which was important for labor.
Asunto(s)
Bifidobacterium longum/inmunología , Suplementos Dietéticos , Microbioma Gastrointestinal/inmunología , Lactobacillus delbrueckii/inmunología , Probióticos/administración & dosificación , Streptococcus thermophilus/inmunología , Adulto , Bifidobacterium longum/genética , Estudios de Casos y Controles , Heces/microbiología , Femenino , Microbioma Gastrointestinal/genética , Expresión Génica , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Humanos , Inmunidad Innata , Interferón gamma/genética , Interferón gamma/inmunología , Interleucina-12/genética , Interleucina-12/inmunología , Interleucina-1beta/genética , Interleucina-1beta/inmunología , Interleucina-2/genética , Interleucina-2/inmunología , Interleucina-5/genética , Interleucina-5/inmunología , Interleucina-6/genética , Interleucina-6/inmunología , Lactobacillus delbrueckii/genética , Aprendizaje Automático , Embarazo , Tercer Trimestre del Embarazo , Streptococcus thermophilus/genética , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
In this study, an immunologically active novel microparticulate mushroom ß-glucan (PRA-1p) was prepared using an alkali-soluble glucan PRA-1 by an emulsification and cross-linking method. PRA-1 was a hyperbranched (1â3),(1â6)-ß-d-glucan with a degree of branching of 0.89, isolated from the sclerotia of Polyporus rhinocerus. PRA-1 had a rod-like conformation, while PRA-1p exhibited a monodisperse and homogeneous spherical conformation with a diameter ranging from 0.3 to 2.0 µm in water. PRA-1p significantly induced nitric oxide and reactive oxygen species production as well as morphological changes of murine macrophages (RAW 264.7 cells) and upregulated their phagocytic activity. Furthermore, PRA-1p treatment markedly enhanced the secretion of cytokines, including cutaneous T cell-attracting chemokine 27, granulocyte-colony-stimulating factor, monocyte chemoattractant protein 1, macrophage inflammatory protein 1α, macrophage inflammatory protein 2, regulated on activation, normal T cell expressed and secreted, soluble tumor necrosis factor receptor 1, and tissue inhibitors of metalloproteinases. Activation of RAW 264.7 cells triggered by PRA-1p was associated with activation of inducible nitric oxide synthase, nuclear factor κB, extracellular signal-regulated kinase, and protein kinase B. This work suggests that novel PRA-1p derived from the mushroom sclerotia of P. rhinocerus has potential application as an immunostimulatory agent.
Asunto(s)
Factores Inmunológicos/química , Factores Inmunológicos/farmacología , Extractos Vegetales/química , Extractos Vegetales/farmacología , Polyporus/química , beta-Glucanos/química , beta-Glucanos/farmacología , Animales , Quimiocina CCL27/genética , Quimiocina CCL27/inmunología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Factores Inmunológicos/aislamiento & purificación , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Ratones , FN-kappa B/genética , FN-kappa B/inmunología , Óxido Nítrico/inmunología , Fagocitosis/efectos de los fármacos , Extractos Vegetales/aislamiento & purificación , Polyporus/inmunología , Células RAW 264.7 , Especies Reactivas de Oxígeno/inmunología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , beta-Glucanos/aislamiento & purificaciónRESUMEN
The aim of the present study was chemical clarification of in vitro Peyer's patch-immunomodulating polysaccharides in sugar cane molasses, and evaluation of in vivo modulating activity on immune function of T lymphocytes in Peyer's patches and on microenvironment of hemopoietic system. Five kinds of glucans, comprising of dextranase-sensitive and activity-related d-glucosyl moieties, were purified as in vitro Peyer's patch-immunomodulating polysaccharides from the molasses. Oral administration of a glucan-enriched subfraction induced IL-2 and GM-CSF-producing T lymphocytes in Peyer's patches, resulting in enhancement of IL-6 production in a hemopoietic microenvironment to boost neutrophil numbers in the peripheral blood stream. Oral administration of purified glucan or glucan-enrich sub-fraction of sugar cane reduced the number of Plasmodium berghei- or P. yoelii-infected erythrocytes in a murine infection model, using polysaccharide alone or via co-administration with the antimalarial drug, artesunate. These results suggested that Peyer's patch-immunomodulating glucans enhanced protective immunity through axis of Peyer's patches-hemopoietic system.
Asunto(s)
Glucanos/farmacología , Hematopoyesis/efectos de los fármacos , Factores Inmunológicos/farmacología , Malaria/tratamiento farmacológico , Ganglios Linfáticos Agregados/efectos de los fármacos , Saccharum/química , Administración Oral , Animales , Células de la Médula Ósea/efectos de los fármacos , Células de la Médula Ósea/inmunología , Femenino , Expresión Génica/efectos de los fármacos , Glucanos/química , Glucanos/aislamiento & purificación , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Hematopoyesis/inmunología , Humanos , Factores Inmunológicos/química , Factores Inmunológicos/aislamiento & purificación , Interleucina-2/genética , Interleucina-2/inmunología , Interleucina-6/genética , Interleucina-6/inmunología , Malaria/genética , Malaria/inmunología , Malaria/parasitología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos ICR , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Ganglios Linfáticos Agregados/inmunología , Extractos Vegetales/química , Plasmodium berghei/efectos de los fármacos , Plasmodium berghei/crecimiento & desarrollo , Plasmodium yoelii/efectos de los fármacos , Plasmodium yoelii/crecimiento & desarrollo , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunologíaRESUMEN
The ability to generate large numbers of distinct types of human dendritic cells (DCs) in vitro is critical for accelerating our understanding of DC biology and harnessing them clinically. We developed a DC differentiation method from human CD34+ precursors leading to high yields of plasmacytoid DCs (pDCs) and both types of conventional DCs (cDC1s and cDC2s). The identity of the cells generated in vitro and their strong homology to their blood counterparts were demonstrated by phenotypic, functional, and single-cell RNA-sequencing analyses. This culture system revealed a critical role of Notch signaling and GM-CSF for promoting cDC1 generation. Moreover, we discovered a pre-terminal differentiation state for each DC type, characterized by high expression of cell-cycle genes and lack of XCR1 in the case of cDC1. Our culture system will greatly facilitate the simultaneous and comprehensive study of primary, otherwise rare human DC types, including their mutual interactions.
Asunto(s)
Linaje de la Célula/inmunología , Células Dendríticas/inmunología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Péptidos y Proteínas de Señalización Intercelular/genética , Proteínas de la Membrana/genética , Receptor Notch1/genética , Antígenos CD34/genética , Antígenos CD34/inmunología , Proteínas de Unión al Calcio , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/inmunología , Diferenciación Celular/efectos de los fármacos , Células Dendríticas/citología , Células Dendríticas/efectos de los fármacos , Expresión Génica , Perfilación de la Expresión Génica , Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Humanos , Imidazoles/farmacología , Inmunofenotipificación , Péptidos y Proteínas de Señalización Intercelular/inmunología , Lipopolisacáridos/farmacología , Proteínas de la Membrana/inmunología , Poli I-C/farmacología , Cultivo Primario de Células , Receptor Notch1/inmunología , Receptores Acoplados a Proteínas G/deficiencia , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/inmunología , Transducción de Señal , Análisis de la Célula IndividualRESUMEN
Oxyresveratrol and its glycoside are important natural active materials. As an effective tyrosine kinase inhibitor, oxyresveratrol may prevent herpes virus infection, inflammation and oxidative stress, as well as protect nerves. In addition, it is known to inhibit cell apoptosis following cerebral ischemia. In recent years, oxyresveratrol and its glycoside have been widely investigated, and their useful biological activities have been explored, indicating that they may be worthy of further comprehensive research. The aim of the present study was to evaluate the photoprotective effects of oxyresveratrol and its ability to abrogate inflammation and oxidative stress in a rat model of spinal cord injury (SCI). The authors identified that oxyresveratrol significantly reversed the SCIinduced inhibition of Basso, Beattie, and Bresnahan scores, inhibited the SCImediated increase in spinal cord water content, significantly suppressed SCIinduced nuclear factorκB/p65, tumor necrosis factorα, interleukin (IL)1ß and IL6 activities and reversed the malondialdehyde, superoxide dismutase, glutathione (GSH) and GSH peroxidase activities in SCI rats. SCIinduced granulocytemacrophage colonystimulating factor (GMCSF), inducible nitric oxide synthase (iNOS) and cyclooxygenase2 (COX2) protein expression was significantly suppressed by oxyresveratrol, and SCImediated inhibition of nuclear factor (erythroidderived 2)like 2 (Nrf2) protein expression was significantly increased by oxyresveratrol. In conclusion, these results suggest that the effects of oxyresveratrol restores SCI, and abrogates inflammation and oxidative stress in rat model of SCI via the GMCSF, iNOS, COX2 and Nrf2 signaling pathway.
Asunto(s)
Antioxidantes/farmacología , Locomoción/efectos de los fármacos , Fármacos Neuroprotectores/farmacología , Extractos Vegetales/farmacología , Traumatismos de la Médula Espinal/tratamiento farmacológico , Médula Espinal/efectos de los fármacos , Estilbenos/farmacología , Animales , Modelos Animales de Enfermedad , Esquema de Medicación , Femenino , Regulación de la Expresión Génica , Glutatión Peroxidasa/genética , Glutatión Peroxidasa/metabolismo , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Inflamación , Inyecciones Intraperitoneales , Interleucina-1beta/antagonistas & inhibidores , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/antagonistas & inhibidores , Interleucina-6/genética , Interleucina-6/metabolismo , Malondialdehído/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Estrés Oxidativo , Ratas , Ratas Sprague-Dawley , Médula Espinal/metabolismo , Médula Espinal/patología , Traumatismos de la Médula Espinal/genética , Traumatismos de la Médula Espinal/metabolismo , Traumatismos de la Médula Espinal/patología , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Factor de Transcripción ReIA/antagonistas & inhibidores , Factor de Transcripción ReIA/genética , Factor de Transcripción ReIA/metabolismo , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
We studied in vivo modifying effect of autotransfusion of human bone marrow mesenchymal stromal cells on ROS generation and production of cytokines (TNFα,TNFß, IL-1α, IL-10, IFNγ, and GM-CSF) and PGE2 by mononuclear cells of patients (N=21) with chronic heart failure. These parameters were evaluated prior to (control) and after (immediately and on day 14) intravenous administration of stromal cells in doses of 100-200×106. Immediately after autotransfusion, significant increase of in vitro zymosan-induced chemiluminescence of blood mononuclear cells from 10 patients was observed. At later terms after autotransfusion (day 14), inhibition of chemiluminescent activity of blood mononuclear cells was revealed in 50% patients. We discuss possible mechanisms of involvement of transplanted autologous bone marrow mesenchymal stromal cells in reprogramming of blood mononuclear phagocytes from the pro- to anti-inflammatory phenotype under conditions of their in vivo interaction manifesting in transition from activation to inhibition of ROS-producing activity of macrophages and significant suppression of in vitro LPS-induced production of TNFα and GM-CSF by blood mononuclears against the background of significantly elevated TNFß, IL-10, and IL-1α concentrations.
Asunto(s)
Insuficiencia Cardíaca/terapia , Leucocitos Mononucleares/inmunología , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/inmunología , Especies Reactivas de Oxígeno/inmunología , Dinoprostona/inmunología , Dinoprostona/metabolismo , Regulación de la Expresión Génica , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Insuficiencia Cardíaca/genética , Insuficiencia Cardíaca/inmunología , Insuficiencia Cardíaca/patología , Humanos , Interferón gamma/genética , Interferón gamma/inmunología , Interleucina-10/genética , Interleucina-10/inmunología , Interleucina-1alfa/genética , Interleucina-1alfa/inmunología , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/patología , Lipopolisacáridos/farmacología , Linfotoxina-alfa/genética , Linfotoxina-alfa/inmunología , Células Madre Mesenquimatosas/citología , Cultivo Primario de Células , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal , Trasplante Autólogo , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Antimalarial drug resistance is the main therapeutic challenge to the control of the disease, making the search for new compounds as alternative treatments of central importance. Propolis has a long history of medicinal use due to its antifungal, antibacterial and antiprotozoal properties. The present study therefore aimed to evaluate the antimalarial activity of the Saudi propolis methanolic extract against Plasmodium chabaudi infection in mice. To this end, albino mice were divided into five groups: the first group was the normal control; the second, third, fourth and fifth groups were infected intraperitoneally with 106 P. chabaudi-parasitized erythrocytes. The last three groups of mice were gavaged with 100 µl of propolis extract (PE) at a dose of 25, 50 and 100 mg PE/kg, respectively, once daily for 7 days. PE significantly suppressed the parasitaemia and showed significant efficacy in ameliorating anaemic conditions in P. chabaudi-infected mice in a dose-dependent manner. Histological investigation of the spleen tissue of treated and untreated mice further supports the antimalarial potential of PE. In addition, our study proved that Saudi PE reduced oxidative damage by decreasing the malondialdehyde (MDA) and increasing the catalase (CAT) activity and the glutathione (GSH) levels. Also, Saudi PE increased the level of some pro-inflammatory cytokines such as IFN-γ, TNF-α, GM-CSF and G-CSF, with the most effective dose being 100 mg PE/kg. In conclusion, PE showed antimalarial and antioxidant activities and provided protection against spleen tissue damage in P. chabaudi-infected mice.
Asunto(s)
Antimaláricos/administración & dosificación , Malaria/tratamiento farmacológico , Extractos Vegetales/administración & dosificación , Plasmodium chabaudi/efectos de los fármacos , Própolis/administración & dosificación , Sustancias Protectoras/administración & dosificación , Bazo/efectos de los fármacos , Animales , Femenino , Glutatión/metabolismo , Factor Estimulante de Colonias de Granulocitos/genética , Factor Estimulante de Colonias de Granulocitos/metabolismo , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Humanos , Malaria/genética , Malaria/metabolismo , Malaria/parasitología , Malondialdehído/metabolismo , Ratones , Parasitemia/tratamiento farmacológico , Parasitemia/genética , Parasitemia/metabolismo , Parasitemia/parasitología , Plasmodium chabaudi/fisiología , Arabia Saudita , Bazo/metabolismo , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Paeonia lactiflora root (baishao in Chinese) is a commonly used herb in traditional Chinese medicines (TCM). Two isomers, paeoniflorin (PF) and albiflorin (AF), are isolated from P. lactiflora. The present study aimed to investigate the protective effects of PF and AF on myelosuppression induced by chemotherapy in mice and to explore the underlying mechanisms. The mouse myelosuppression model was established by intraperitoneal (i.p.) injection of cyclophosphamide (CP, 200 mg·kg(-1)). The blood cell counts were performed. The thymus index and spleen index were also determined and bone morrow histological examination was performed. The levels of tumor necrosis factor-α (TNF-α) in serum and colony-stimulating factor (G-CSF) in plasma were measured by Enzyme-Linked Immunosorbent Assays (ELISA) and the serum levels of interleukin-3 (IL-3), granulocyte-macrophagecolony-stimulatingfactor (GM-CSF), and interleukin-6 (IL-6) were measured by radioimmunoassay (RIA). The levels of mRNA expression protein of IL-3, GM-CSF and G-CSF in spleen and bone marrow cells were determined respectively. PF and AF significantly increased the white blood cell (WBC) counts and reversed the atrophy of thymus. They also increased the serum levels of GM-CSF and IL-3 and the plasma level of G-CSF and reduced the level of TNF-α in serum. PF enhanced the mRNA level of IL-3 and AF enhanced the mRNA levels of GM-CSF and G-CSF in the spleen. PF and AF both increased the protein levels of GM-CSF and G-CSF in bone marrow cells. In conclusion, our results demonstrated that PF and AF promoted the recovery of bone marrow hemopoietic function in the mouse myelosuppression model.
Asunto(s)
Antineoplásicos/efectos adversos , Hidrocarburos Aromáticos con Puentes/administración & dosificación , Ciclofosfamida/efectos adversos , Medicamentos Herbarios Chinos/administración & dosificación , Glucósidos/administración & dosificación , Enfermedades Hematológicas/prevención & control , Monoterpenos/administración & dosificación , Paeonia/química , Animales , Factor Estimulante de Colonias de Granulocitos/genética , Factor Estimulante de Colonias de Granulocitos/metabolismo , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Enfermedades Hematológicas/etiología , Enfermedades Hematológicas/genética , Enfermedades Hematológicas/metabolismo , Humanos , Interleucina-3/genética , Interleucina-3/metabolismo , Interleucina-6/metabolismo , Masculino , Ratones , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Granulocyte macrophage-colony stimulating factor (GM-CSF) is a potent immunomodulatory cytokine that is known to facilitate vaccine efficacy by promoting the development and prolongation of both humoral and cellular immunity. In the past years we have generated a novel codon-optimized GM-CSF gene as an adjuvant. The codon-optimized GM-CSF gene significantly increased protein expression levels in all cells tested and helped in generating a strong immune responses against HIV-1 Gag and HPV-associated cancer. Here, we review the literature dealing with the adjuvant activity of GM-CSF both in animal models and clinical trials. We anticipate that the codon-optimized GM-CSF gene offers a practical molecular strategy for potentiating immune responses to tumor cell-based vaccinations as well as other immunotherapeutic strategies.
Asunto(s)
Vacunas contra el SIDA/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Vacunas contra el Cáncer/inmunología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/administración & dosificación , Vacunas contra el SIDA/administración & dosificación , Adyuvantes Inmunológicos/genética , Animales , Vacunas contra el Cáncer/administración & dosificación , Ensayos Clínicos como Asunto , Evaluación Preclínica de Medicamentos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Humanos , Modelos AnimalesRESUMEN
This study was designed to investigate the protective effects of the centella triterpene saponins (EXT) on cyclophosphamide (CYP)-induced hepatotoxicity and immunosuppression in rats. The phytochemical profile of EXT was analyzed for centella saponins by using high-performance liquid chromatographic (HPLC). Therapeutic efficacy of EXT (250 mg/kg/day p.o) on hematological profile of blood, liver function markers, and cytokine profiles in CYP (10 mg/kg/day p.o)-treated rats. In addition, weights of immune organs (spleen and thymus) and histopathological changes in the liver, intestine, and spleen were also evaluated. The active principles in EXT were identified as madecassoside, asiaticoside, and asiatic acid by HPLC analysis. Upon administration of EXT, enhanced levels of glutamate pyruvate transaminase, alkaline phosphatase, and lipid peroxidation were found reduced while the levels of reduced glutathione and hematological parameters and relative weights of immune organs were restored to normal in CYP-treated rats. The hepatic mRNA level of TNF-α, which was increased during CYP administration, was significantly decreased by the EXT treatment. The decreased levels of mRNA expression of other cytokines like IFN-γ, IL-2, GM-CSF, after CYP treatment, were also found elevated upon administration of the EXT. Histopathological examination of the intestine, liver, and spleen indicated that the extract could attenuate the CYP-induced hepatic and immune organ damage. These results indicated that EXT modulated the immune and hepatic system function of rats against CYP-induced immunosuppression and hepatotoxicity by restoring the cytokine production, antioxidant system, and multiorgan injury. Thus, triterpene saponins may provide protective and/or therapeutic alternative against the immune-mediated liver diseases.
Asunto(s)
Antineoplásicos Alquilantes/toxicidad , Ciclofosfamida/toxicidad , Hígado/efectos de los fármacos , Extractos Vegetales/farmacología , Saponinas/farmacología , Triterpenos/farmacología , Fosfatasa Alcalina/metabolismo , Animales , Centella/química , Citoprotección , Evaluación Preclínica de Medicamentos , Recuento de Eritrocitos , Expresión Génica/efectos de los fármacos , Expresión Génica/inmunología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Interferón gamma/genética , Interferón gamma/metabolismo , Interleucina-2/genética , Interleucina-2/metabolismo , Intestinos/efectos de los fármacos , Intestinos/patología , Recuento de Leucocitos , Peroxidación de Lípido , Hígado/patología , Masculino , Tamaño de los Órganos/efectos de los fármacos , Recuento de Plaquetas , Ratas Wistar , Bazo/efectos de los fármacos , Bazo/patología , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
1. The potential use of CpG oligodeoxynucleotides and/or Astragalus polysaccharide (APS) as adjuvants for the culture of chicken bone-marrow-derived dendritic cells (chBM-DCs) was investigated. 2. Chicken dendritic cells (DCs) were isolated and cultured in the presence of recombinant chicken granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4. The chBM-DC displayed typical DC morphology and expressed DC surface markers (MHC-II and CD11c). 3. Cultured chBM-DC showed effective T-cell activation in vitro, based on a mixed lymphocyte response (MLR). Flow cytometry analysis showed an increased proportion of cells expressing CD40 and CD80 in the APS-stimulated culture, compared to the control culture. In the MLR, the APS- and CpG-stimulated chBM-DC could activate T-cells more than control chBM-DC. Real-time PCR assays showed that CpG can activate the TLR21 and an inflammatory response, while APS just reduced the expression of IRF-3. 4. The results demonstrated that in vitro the adjuvant CpG can stimulate chBM-DC to mature by activation of the TLR-signalling pathway, whereas the adjuvant APS stimulates maturation of chBM-DC in vitro to a lesser degree and by another mechanism.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Planta del Astrágalo/química , Células de la Médula Ósea/efectos de los fármacos , Pollos , Células Dendríticas/efectos de los fármacos , Oligodesoxirribonucleótidos/farmacología , Polisacáridos/farmacología , Animales , Proteínas Aviares/genética , Proteínas Aviares/metabolismo , Células Cultivadas , Células Dendríticas/inmunología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Interleucina-4/genética , Interleucina-4/metabolismo , Extractos Vegetales/farmacología , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMEN
A water soluble extract from the medicinal mushroom Agrocybe aegerita has been shown to stimulate splenocyte proliferation, cytotoxic activity, and tumor rejection effect in tumor-bearing mouse models. In the present study, the crude extract was separated into a protein component fraction (Yp), mainly containing lectins and serine proteinase, and a small molecule component fraction (Ys), mainly containing triethylene glycol, α-bisabolol, n-hexadecanoic acid, and so on. The antitumor activity of the fractions was investigated in a tumor-bearing BALB/c mouse model. Repeat administration of Yp and Ys significantly inhibited tumor growth (P<.001), but little toxicity was observed. Moreover, the protein fraction Yp performed better than Ys in both antitumor and lifespan-prolonging activity. The cytokine expression levels in serum and splenocytes from extract-treated mice were selectively screened by enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction, and the results showed that Yp upregulated the mRNA level of Th2 cytokine interleukin-10 (P<.01), and Ys increased the mRNA level of granulocyte-macrophage colony-stimulating factor (P<.01) and anti-inflammatory cytokine transforming growth factor-ß (P<.01). All these data suggest that Yp and Ys can inhibit tumor growth via different mechanisms, which promotes the understanding of antitumor properties of medicinal fungi.
Asunto(s)
Agrocybe/química , Antineoplásicos Fitogénicos/administración & dosificación , Citocinas/genética , Neoplasias/tratamiento farmacológico , Extractos Vegetales/administración & dosificación , Proteínas de Plantas/administración & dosificación , Plantas Medicinales/química , Verduras/química , Animales , Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/aislamiento & purificación , Citocinas/metabolismo , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Humanos , Interleucina-10/genética , Interleucina-10/metabolismo , Ratones , Ratones Endogámicos BALB C , Neoplasias/genética , Neoplasias/metabolismo , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Proteínas de Plantas/química , Proteínas de Plantas/aislamiento & purificación , Factor de Crecimiento Transformador beta1/genética , Factor de Crecimiento Transformador beta1/metabolismoRESUMEN
BACKGROUND: An adenovirus that expresses both interleukin (IL)-12 and granulocyte-macrophage colony-stimulating-factor (GM-CSF) has been proven to be very effective in treating several tumors, but causes serious normal tissue toxicities. METHODS: In this study, a novel adenoviral vector was constructed by placing the human GM-CSF gene under the control of the CMV-IE promoter and human IL-12 gene under the control of heat shock protein 70B gene promoter. Both hGM-CSF and hIL-12 expressions in virus-infected tumor cells were analyzed in vitro and in vivo when underlying single or multiple rounds of hyperthermia. RESULTS: We observed constitutive high expression of human GM-CSF and heat-induced expression of human IL-12 after a single round of hyperthermia post viral infection. The heat-induced hIL-12 expression exhibited a pulse-like pattern with a peak at 24 hrs followed by a decline 48 hrs post heat stress. Repeated heat treatment was more effective in inducing hIL-12 expression than a one-time heat treatment. Interestedly, we also observed that constitutive expression of hGM-CSF could be stimulated by heat stress in tested tumor cells. CONCLUSION: Our study provided a novel strategy for combined gene therapy that allows constitutive expression of a non-toxic gene such as GM-CSF and heat-induced expression of a toxic gene such as IL-12. In addition, our study also showed that hyperthermia can be used to trigger gene expression in temporal and special manner.
Asunto(s)
Terapia Genética/métodos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Hipertermia Inducida/métodos , Interleucina-12/genética , Adenoviridae/genética , Animales , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/terapia , Carcinoma de Pulmón de Células no Pequeñas/virología , Técnicas de Cultivo de Célula , Línea Celular Tumoral , Factor Estimulante de Colonias de Granulocitos y Macrófagos/biosíntesis , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacocinética , Humanos , Interleucina-12/biosíntesis , Interleucina-12/farmacocinética , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/terapia , Neoplasias Hepáticas/virología , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/terapia , Neoplasias Pulmonares/virología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Granulocyte-macrophage colony-stimulating factor (GMCSF) and MCP3 (aka CCL7) exert complementary, nonoverlapping, proimmune effects on responsive lymphoid and myeloid cells. We hypothesized that a synthetic cytokine linking GMCSF to MCP3 (hereafter GMME3) as part of a single polypeptide would acquire novel, therapeutically desirable immunomodulatory properties. We demonstrate that GMME3 has enhanced CC-chemokine receptor (CCR)-mediated intracellular Ca(++) mobilization with selective effects on the CD21(hi)CD24(hi) CD1.d(hi) subset of splenic B cells inducing substantial interleukin 10 (IL10) production. We demonstrate that B(GMME3) exert their suppressive effect through an IL10-mediated inhibition of antigen presentation. More importantly, B(GMME3) inhibit the reactivation of encephalomyelitis (EAE)-derived or TGFß/IL6 differentiated Th17 cells by altering their polarization toward a Th1 or Th2 phenotype. The secretion of interferon-γ (IFNγ) and IL4 in turn inhibits IL17 production. The adoptive transfer of B(GMME3), but not IL10(-/-) B(GMME3) cells, to mice symptomatic with experimental autoimmune encephalitis significantly improves their disease score and inhibits lymphoid infiltration into the central nervous system (CNS). We propose that designed CCR modulators such as GMME3, allows for conversion of naive B-cells to a novel suppressor phenotype allowing for the personalized cell therapy of autoimmune ailments.
Asunto(s)
Linfocitos B/inmunología , Encefalomielitis Autoinmune Experimental/terapia , Inmunoterapia , Inflamación/terapia , Interleucina-10/inmunología , Células Th17/inmunología , Traslado Adoptivo , Animales , Presentación de Antígeno , Linfocitos B/metabolismo , Calcio/inmunología , Calcio/metabolismo , Diferenciación Celular , Sistema Nervioso Central/inmunología , Sistema Nervioso Central/patología , Quimiocina CCL7/genética , Quimiocina CCL7/inmunología , Encefalomielitis Autoinmune Experimental/genética , Encefalomielitis Autoinmune Experimental/inmunología , Encefalomielitis Autoinmune Experimental/patología , Femenino , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Células HEK293 , Humanos , Inmunomodulación , Inflamación/genética , Inflamación/inmunología , Inflamación/patología , Interleucina-10/biosíntesis , Ratones , Ratones Transgénicos , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Bazo/inmunología , Bazo/patología , Células Th17/metabolismoRESUMEN
Fungal ß-glucan is a representative pathogen-associated microbial pattern (PAMP) from mushroom, yeast, and fungi, and stimulates innate as well as acquired immune systems. It is a widely used functional food to enhance immunity. Such plant extracts have been known as folk medicines and reported to show various biological activities beneficial to human health, such as anti-tumor, anti-allergic, and anti-inflammatory activities. In the present study, the cooperative effect of bamboo water-soluble methanol precipitation (BWMP), a macromolecular fraction of the hot-water extract of Sasa veitchii (Japanese folk medicine Kumazasa), and the ß-glucan from the medicinal mushroom Sparassis crispa (SCG) was analyzed in vitro using DBA/2 mice. The splenocytes from male DBA/2 mice were cultured with BWMP in the presence of SCG, and the responses were assessed by measuring cytokines. BWMP suppressed IFN-γ and GM-CSF production by SCG, but not TNF-α production. To analyze the specificity of the reaction, similar experiments were conducted with BWMP in the presence of bacterial lipopolysaccharide (LPS); however, none of the cytokines were inhibited. Cytokine production of splenocytes by SCG was suggested to be largely dependent on the binding of lymphocytes with dendritic cells. Functions of BWMP were also analyzed by mixed lymphocyte reaction, and IFN-γ production was suppressed. These findings suggested that BWMP modulated the cell-to-cell contact induced by SCG and inhibited cytokine production. It is strongly suggested that the plant extracts modulate the immunostimulating effects of medicinal mushrooms. Cooperative effects of plants and mushrooms would be an important issue for functional foods.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Agaricales/química , Extractos Vegetales/farmacología , Sasa/química , beta-Glucanos/farmacología , Adyuvantes Inmunológicos/química , Animales , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Relación Dosis-Respuesta a Droga , Regulación de la Expresión Génica , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Japón , Masculino , Medios de Comunicación de Masas , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Extractos Vegetales/química , Bazo/citología , Bazo/efectos de los fármacos , beta-Glucanos/química , beta-Glucanos/metabolismoRESUMEN
CCR2 is a chemokine receptor widely expressed by lymphomyeloid cells involved in maladaptive autoimmune ailments. Therefore CCR2 is of great interest as a biological target for immune suppression due to its direct implication in autoimmune diseases such as rheumatoid arthritis. We have generated a novel fusion protein using GM-CSF and an N-terminal truncated version of MCP-1/CCL2 (6-76, GMME1) and investigated its utility as a CCR2-specific immune suppressor. Using BRET studies, we found that distinct to CCL2, GMME1 binding to CCR2 led to altered conformational changes in the CCR2 homodimer and did not induce the recruitment of beta-arrestin 2 to the receptor. However, CCR2-dependent calcium mobilization, BAX induction and caspase-3 activation followed by cell death was observed. Using Th17 cells harvested from DBA/1 mice ill with bovine collagen-induced arthritis, we demonstrate that GMME1 is capable of blocking their production of IL-17 in vitro. Upon its delivery to mice symptomatic with inflammatory arthritis, a robust clinical recovery occurred with decreased paw thickness to normal levels and a significant reduction in anti-collagen Ab titer and rheumatoid factor titer, as well as reduction of proinflammatory cytokines levels both intraarticular and systemic. Our data demonstrate that GMME1 is a powerful synthetic suppressor cytokine that coopts CCR2-dependent cellular signaling and blunts the effects of CCR2-expressing lymphomyeloid cells causative of autoimmune arthritis.
Asunto(s)
Artritis Experimental/tratamiento farmacológico , Quimiocina CCL2/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Ingeniería de Proteínas/métodos , Receptores CCR2/metabolismo , Proteínas Recombinantes de Fusión/farmacología , Animales , Artritis Experimental/prevención & control , Células Cultivadas , Modelos Animales de Enfermedad , Inflamación/tratamiento farmacológico , Inflamación/prevención & control , Interleucina-17 , Ratones , Unión Proteica , Receptores CCR2/química , Proteínas Recombinantes de Fusión/uso terapéutico , Subgrupos de Linfocitos TRESUMEN
AIM OF THE STUDY: We aimed to demonstrate ethnopharmacological role of Codonopsis lanceolata L., known to be used as a folk qi tonic herb to aid those who are weakened by blood loss or other injury in Korea, by examining the expressional level of hematopoietic cytokines. RESULTS: BuOH fraction strongly up-regulated mRNA expression of granulocyte-macrophage-colony stimulating factor (GM-CSF). Similarly, BuOH fraction also increased the proliferation of splenocytes. According to the analysis of transcription factors and their regulatory signaling enzymes by using reporter gene assay, pharmacological dissection and Western blotting, NF-kappaB and AP-1 were found to be up-regulated by BuOH fraction. Furthermore, ERK and PI3K seemed to be regarded as positive upstream signaling enzymes responsible for activation of these transcription factors. CONCLUSION: These data suggest that the enthopharmacological role of Codonopsis lanceolata on the recovery of blood loss and spleen qi may be due to its up-regulatory effect of GM-CSF expression via activating relevant signaling cascades.
Asunto(s)
Codonopsis/química , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Macrófagos/efectos de los fármacos , Animales , Secuencia de Bases , Western Blotting , Línea Celular , Cartilla de ADN , Evaluación Preclínica de Medicamentos , Expresión Génica/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , FN-kappa B/metabolismo , ARN Mensajero/genética , Factor de Transcripción AP-1/metabolismo , Regulación hacia ArribaRESUMEN
BACKGROUND: Green Coffea arabica L. seed oil is being widely used in cosmetic formulations, although its effects on human skin cells are not clear and most observations are unpublished. AIMS: In this study, we evaluated the in vitro effects of green coffee (C. arabica L.) oil (GCO) on the synthesis of collagen, elastin, and glycosaminoglycans (GAG) and in the release of transforming growth factor-beta1 (TGF-beta1) and granulocyte-macrophage colony-stimulating factor (GM-CSF) by human skin fibroblasts. We also investigated the ability of GCO to increase aquaglycerolporins-3 (AQP-3) mRNA expression in cultured keratinocytes and human skin explants. METHODS: Human fibroblasts were incubated for 48 h with several GCO concentrations (3.12, 6.25, 12.5, 25.0 and 50.0 mg/mL). The levels of growth factors and extracellular matrix compounds in the culture supernatant were measured using commercial kits. To evaluate AQP-3 relative expression, using real-time reverse transcription polymerase chain reaction, keratinocytes were incubated for 3-6 h with the GCO optimal concentration of 25.0 mg/mL. Histological sections of human skin were also incubated with GCO (25.0 mg/mL) and immunostained by antiserum against AQP-3. RESULTS: Our results demonstrated that incubation with GCO produces a dose-dependent stimulation in the synthesis of collagen, elastin, and GAG, in addition to increasing the release of the growth factors TGF-beta1 and GM-CSF. GCO also induced the expression of AQP-3 mRNA, which reached levels up to 6.5-fold higher than those of the control cultures. CONCLUSION: The findings presented herein suggest that GCO might improve physiological balance in the skin, thus allowing the formation of new connective tissue, and preventing epidermis dryness by increasing AQP-3 levels. Taking into account the limitations of in vitro studies, it is encouraging in this context to consider CGO as an adjuvant to be used in dermocosmetic formulations. Clinical studies are in progress in our laboratory aiming to further investigate the protective effects of CGO in the skin.