ABSTRACT
Dendritic cells (DCs) are play critical roles in the priming and regulation of immune responses. DCs rapidly process and convey these antigens to prime antigen-specific T cells. Therefore, regulation of DCs functions is important for immunity and immunotherapies. Immune adjuvants for DCs activation are needed to improve the efficacy of vaccines against tumors and many infectious diseases. Therefore, we demonstrate that H. fusiformis extract can regulate DCs maturation and activation. H. fusiformis extract induced costimulatory molecules (CD 80 and CD86), antigen-presenting molecules (major histocompatibility complex (MHC) I and II), CCR7 expression, and interleukin (IL)-12 production in DCs. These effects are associated with upregulation of mitogen-activated protein kinase (MAPK) signaling pathway. In addition, H. fusiformis extract induces costimulatory molecules on splenic DCs and activated CD8+ T cells in vivo. Taken together, these findings suggest that H. fusiformis extract may be a potential efficient immune therapeutic compound in DCs-mediated immunotherapies. ABBREVIATIONS: CTL: cytotoxic T lymphocytes; DCs: dendritic cells; ERK: extracellular signal-regulated kinases; IL: interleukini; JNK: c-Jun N-terminal kinase; MAPK: mitogen-activated protein kinase; MHC: major histocompatibility complex.
Subject(s)
Dendritic Cells/cytology , Dendritic Cells/drug effects , Plant Extracts/pharmacology , Sargassum/chemistry , Cell Differentiation/drug effects , Cell Line , Dendritic Cells/immunology , Dendritic Cells/metabolism , Gene Expression Regulation/drug effects , Humans , Interleukin-12/biosynthesis , Lymphocyte Activation/drug effects , MAP Kinase Signaling System/drug effects , Receptors, CCR7/metabolismABSTRACT
Immune-checkpoint blockers can promote sustained clinical responses in a subset of cancer patients. Recent research has shown that a subpopulation of tumor-infiltrating dendritic cells functions as gatekeepers, sensitizing tumors to anti-PD-1 treatment via production of interleukin-12 (IL-12). Hypothesizing that myeloid cell-targeted nanomaterials could be used to deliver small-molecule IL-12 inducers, we performed high-content image-based screening to identify the most efficacious small-molecule compounds. Using one lead candidate, LCL161, we created a myeloid-targeted nanoformulation that induced IL-12 production in intratumoral myeloid cells in vivo, slowed tumor growth as a monotherapy, and had no significant systemic toxicity. These results pave the way for developing combination immunotherapeutics by harnessing IL-12 production for immunostimulation.
Subject(s)
Alkynes/pharmacology , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Colonic Neoplasms/therapy , Immunotherapy , Myeloid Cells/drug effects , Oligopeptides/pharmacology , Small Molecule Libraries/pharmacology , Thiazoles/pharmacology , Alkynes/chemistry , Animals , Antineoplastic Agents/chemistry , Cell Line, Tumor , Colonic Neoplasms/metabolism , Colonic Neoplasms/pathology , Dendritic Cells , Drug Carriers/chemistry , Drug Evaluation, Preclinical , Interleukin-12/biosynthesis , Mice , Myeloid Cells/metabolism , Myeloid Cells/pathology , Nanoparticles/chemistry , Oligopeptides/chemistry , Small Molecule Libraries/chemistry , Thiazoles/chemistryABSTRACT
Euglena gracilis Z is a micro-algae that is used as a food or nutritional supplement. Paramylon, the carbohydrate storage substance of Euglena gracilis Z has ß-1, 3-glucan structure. Euglena gracilis Z and paramylon are reported to affect the immune system. In this study, we investigated the protective effects of Euglena gracilis Z and paramylon against influenza virus infection in mice. Euglena gracilis Z and paramylon were administered to mice as a 2% dietary mixture ad libitum. At 2 weeks after initiation of dietary administration, mice were infected intranasally with influenza virus A/PR/8/34 (H1N1). Survival rate was monitored 10 days after infection. In addition, we performed virus titer and cytokine profiles in the lung. High survival rates were observed for Euglena gracilis Z and paramylon-treated groups compared to the control group. Significantly lower virus titer in the lung was observed in the Euglena gracilis Z and paramylon-treated groups compared to the control group from day 1 after infection. Higher amount of IL-1ß, IL-6, IL-12 (p70), IFN-γ, and IL-10 was observed in the paramylon groups compared to the control group. Our data therefore reveals a novel immunoregulatory role of the Euglena gracilis Z and paramylon which provides protection against influenza virus infection.
Subject(s)
Adjuvants, Immunologic/administration & dosage , Dietary Supplements , Euglena gracilis/immunology , Glucans/administration & dosage , Lung/drug effects , Orthomyxoviridae Infections/diet therapy , Administration, Oral , Animals , Euglena gracilis/chemistry , Female , Influenza A Virus, H1N1 Subtype/drug effects , Influenza A Virus, H1N1 Subtype/growth & development , Influenza A Virus, H1N1 Subtype/pathogenicity , Interferon-gamma/biosynthesis , Interferon-gamma/immunology , Interleukin-10/biosynthesis , Interleukin-10/immunology , Interleukin-12/biosynthesis , Interleukin-12/immunology , Interleukin-1beta/biosynthesis , Interleukin-1beta/immunology , Interleukin-6/biosynthesis , Interleukin-6/immunology , Lung/immunology , Lung/virology , Mice , Mice, Inbred BALB C , Orthomyxoviridae Infections/immunology , Orthomyxoviridae Infections/mortality , Orthomyxoviridae Infections/virology , Survival AnalysisABSTRACT
Four sesquiterpene lactones, mikanolide, deoxymikanolide, dihydromikanolide and scandenolide, were isolated by a bioassay-guided fractionation of Mikania variifolia and Mikania micrantha dichloromethane extracts. Mikanolide and deoxymikanolide were the major compounds in both extracts (2.2% and 0.4% for Mikania variifolia and 21.0% and 6.4% for Mikania micrantha respectively, calculated on extract dry weight). Mikanolide, deoxymikanolide and dihydromikanolide were active against Trypanosoma cruzi epimastigotes (50% inhibitory concentrations of 0.7, 0.08 and 2.5 µg/mL, for each compound respectively). These sesquiterpene lactones were also active against the bloodstream trypomastigotes (50% inhibitory concentrations for each compound were 2.1, 1.5 and 0.3 µg/mL, respectively) and against amastigotes (50% inhibitory concentrations for each compound were 4.5, 6.3 and 8.5 µg/mL, respectively). By contrast, scandenolide was not active on Trypanosoma cruzi. Besides, mikanolide and deoxymikanolide were also active on Leishmania braziliensis promastigotes (50% inhibitory concentrations of 5.1 and 11.5 µg/mL, respectively). The four sesquiterpene lactones were tested for their cytotoxicity on THP 1 cells. Deoxymikanolide presented the highest selectivity index for trypomastigotes (SI = 54) and amastigotes (SI = 12.5). In an in vivo model of Trypanosoma cruzi infection, deoxymikanolide was able to decrease the parasitemia and the weight loss associated to the acute phase of the parasite infection. More importantly, while 100% of control mice died by day 22 after receiving a lethal T. cruzi infection, 70% of deoxymikanolide-treated mice survived. We also observed that this compound increased TNF-α and IL-12 production by macrophages, which could contribute to control T. cruzi infection.
Subject(s)
Lactones/pharmacology , Leishmania braziliensis/drug effects , Mikania/chemistry , Plant Extracts/pharmacology , Sesquiterpenes/pharmacology , Trypanosoma cruzi/drug effects , Animals , Chagas Disease/drug therapy , Chagas Disease/parasitology , Drug Discovery , Interleukin-12/biosynthesis , Interleukin-12/immunology , Lactones/administration & dosage , Lactones/chemistry , Lactones/isolation & purification , Lactones/therapeutic use , Life Cycle Stages/drug effects , Male , Mice , Mice, Inbred BALB C , Plant Extracts/chemistry , Sesquiterpenes/chemistry , Sesquiterpenes/isolation & purification , Sesquiterpenes, Germacrane/administration & dosage , Sesquiterpenes, Germacrane/isolation & purification , Sesquiterpenes, Germacrane/pharmacology , Sesquiterpenes, Germacrane/therapeutic use , Trypanosoma cruzi/isolation & purification , Tumor Necrosis Factor-alpha/biosynthesis , Tumor Necrosis Factor-alpha/immunologyABSTRACT
The anti-proliferative agent hexamethylene bisacetamide (HMBA) belongs to a class of hybrid bipolar compounds developed more than 30 y ago for their ability to induce terminal differentiation of transformed cells. Recently, HMBA has also been shown to trigger HIV transcription from latently infected cells, via a CDK9/HMBA inducible protein-1 dependent process. However, the effect of HMBA on the immune response has not been explored. We observed that pretreatment of human peripheral blood mononuclear cells with HMBA led to a markedly increased production of IL-12 and IFN-γ, but not of TNF-α, IL-6, and IL-8 upon subsequent infection with Burkholderia pseudomallei and Salmonella enterica HMBA treatment was also associated with better intracellular bacterial control. HMBA significantly improved IL-12p70 production from CD14+ monocytes during infection partly via the induction of type I IFN in these cells, which primed an increased transcription of the p35 subunit of IL-12p70 during infection. HMBA also increased early type I IFN transcription in human monocytic and epithelial cell lines, but this was surprisingly independent of its previously reported effects on positive transcription elongation factor b and HMBA inducible protein-1. Instead, the effect of HMBA was downstream of a calcium influx, and required the pattern recognition receptor and adaptor STING but not cGAS. Our work therefore links the STING-IRF3 axis to enhanced IL-12 production and intracellular bacterial control in primary monocytes. This raises the possibility that HMBA or related small molecules may be explored as therapeutic adjuvants to improve disease outcomes during intracellular bacterial infections.
Subject(s)
Acetamides/pharmacology , Adjuvants, Immunologic , Interferon Type I/biosynthesis , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/microbiology , Membrane Proteins/metabolism , Acetamides/therapeutic use , Burkholderia pseudomallei/drug effects , Burkholderia pseudomallei/immunology , Cell Line , Cells, Cultured , Cytoplasm/immunology , Cytoplasm/microbiology , Epithelial Cells/drug effects , Epithelial Cells/immunology , Epithelial Cells/microbiology , Humans , Interferon Regulatory Factor-3/metabolism , Interferon Type I/genetics , Interferon Type I/immunology , Interferon-gamma/biosynthesis , Interferon-gamma/immunology , Interleukin-12/biosynthesis , Interleukin-12/immunology , Interleukin-6/biosynthesis , Interleukin-6/immunology , Interleukin-8/biosynthesis , Interleukin-8/immunology , Leukocytes, Mononuclear/immunology , Membrane Proteins/immunology , Nucleotidyltransferases/genetics , Nucleotidyltransferases/metabolism , Salmonella enterica/drug effects , Salmonella enterica/immunology , Signal Transduction , Tumor Necrosis Factor-alpha/biosynthesis , Tumor Necrosis Factor-alpha/immunologyABSTRACT
Cell biology differs between traditional cell culture and 3-dimensional (3-D) systems, and is modulated by the extracellular matrix. Experimentation in 3-D presents challenges, especially with virulent pathogens. Mycobacterium tuberculosis (Mtb) kills more humans than any other infection and is characterised by a spatially organised immune response and extracellular matrix remodelling. We developed a 3-D system incorporating virulent mycobacteria, primary human blood mononuclear cells and collagen-alginate matrix to dissect the host-pathogen interaction. Infection in 3-D led to greater cellular survival and permitted longitudinal analysis over 21 days. Key features of human tuberculosis develop, and extracellular matrix integrity favours the host over the pathogen. We optimised multiparameter readouts to study emerging therapeutic interventions: cytokine supplementation, host-directed therapy and immunoaugmentation. Each intervention modulates the host-pathogen interaction, but has both beneficial and harmful effects. This methodology has wide applicability to investigate infectious, inflammatory and neoplastic diseases and develop novel drug regimes and vaccination approaches.
Subject(s)
Host-Pathogen Interactions/drug effects , Leukocytes, Mononuclear/drug effects , Models, Biological , Mycobacterium tuberculosis/pathogenicity , Spheroids, Cellular/drug effects , Alginates/chemistry , Antigens, Bacterial/pharmacology , Bacterial Proteins/pharmacology , Chemokine CCL2/biosynthesis , Chemokine CCL2/metabolism , Chemokine CXCL10/biosynthesis , Chemokine CXCL10/metabolism , Coculture Techniques , Collagen/chemistry , Dinoprostone/pharmacology , Extracellular Matrix/chemistry , Extracellular Matrix/drug effects , Extracellular Matrix/immunology , Gene Expression Regulation , Glucuronic Acid/chemistry , Granulocyte-Macrophage Colony-Stimulating Factor/biosynthesis , Granulocyte-Macrophage Colony-Stimulating Factor/metabolism , Hexuronic Acids/chemistry , Host-Pathogen Interactions/immunology , Humans , Interleukin-12/biosynthesis , Interleukin-12/metabolism , Interleukin-1beta/biosynthesis , Interleukin-1beta/metabolism , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/microbiology , Microspheres , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/physiology , Spheroids, Cellular/immunology , Spheroids, Cellular/microbiology , VirulenceABSTRACT
The crude polysaccharide (KPV-0) isolated from Korean persimmon vinegar was fractionated using gel filtration chromatography to enhance the immunostimulatory activity and to identify the structural features of active fraction. Among three fractions, KPV-I obtained in a void volume, demonstrated the potent production of macrophage-stimulating mediators, including tumor necrosis factor-α, interleukin (IL)-6, IL-12, and nitric oxide. KPV-I showed a combined single peak with high molecular weight of 55,000Da by high performance size exclusion chromatography. Component sugar analysis revealed that KPV-I contained mainly of arabinose, mannose, galactose, rhamnose and galacturonic acid. Single radial gel diffusion assay using ß-glucosyl Yariv reagent showed that KPV-I contained arabinogalactan protein with 13.7%. Methylation analysis indicated that KPV-I contained 21 kinds of neutral glycosidic linkages, which seemed to be composed three kinds of polysaccharide; that is a rhamnogalacturonan-I (65-70%) derived from persimmon as a raw material, a mannan (20-25%) derived from fermentation-associated microorganisms, and a linear glucans (less than 10%). In conclusion, polysaccharide isolated from persimmon vinegar could augment the macrophage stimulation, and a large amounts of RG-I polysaccharide derived from persimmon is likely a crucial role in expression of the activity in persimmon vinegar.
Subject(s)
Acetic Acid/chemistry , Chemical Fractionation/methods , Diospyros/chemistry , Pectins/pharmacology , Animals , Cell Line , Chromatography, Gel , Hexoses/isolation & purification , Hexoses/pharmacology , Hexuronic Acids/isolation & purification , Hexuronic Acids/pharmacology , Interleukin-12/biosynthesis , Interleukin-12/metabolism , Interleukin-6/biosynthesis , Interleukin-6/metabolism , Macrophage Activation/drug effects , Macrophages/cytology , Macrophages/drug effects , Macrophages/immunology , Mice , Molecular Weight , Nitric Oxide/agonists , Nitric Oxide/biosynthesis , Pectins/chemistry , Pectins/isolation & purification , Republic of Korea , Tumor Necrosis Factor-alpha/biosynthesis , Tumor Necrosis Factor-alpha/metabolismABSTRACT
OBJECTIVE: To determine the mechanisms by which a traditional herbal medicine, Senkinnaidakusan (SKNS), controls Th2 responses, we examined the production of IL-12 by murine macrophages treated with SKNS. RESULTS: Treatment with SKNS significantly increased TLR4 mRNA in macrophages. Furthermore, pre-treatment with SKNS enhanced the production of IL-12 by macrophages stimulated with LPS. When SKNS was orally administered to C3H/HeN mice at the induction phase after OVA sensitization, the serum levels of OVA-specific immunoglobulin (Ig)E and IgG1 decreased, Interleukin (IL)-4 production by spleen T cells in response to OVA was significantly suppressed, while interferon (IFN)-gamma production was increased. After nasal challenge of OVA, eosinophilic infiltration in the nasal mucosa and the number of sneezes were significantly inhibited in SKNS-treated mice compared with control mice. Besides, expression of IL-5 in the nasal mucosa was also inhibited. Using another strain of mice, C3H/HeJ (TLR4 negative), there was no difference in OVA-specific Igs or splenic cytokine production between the SKNS treatment and non-treatment groups. The eosinophilic infiltration in the nasal mucosa, the number of sneezes and IL-5 expression in the nasal mucosa were also not effected even after SKNS treatment. CONCLUSION: These results suggest that oral administration of SKNS inhibits Th2 responses by enhancement of IL-12 release from macrophages via up-regulation of TLR4 expression.
Subject(s)
Interleukin-12/biosynthesis , Macrophages/metabolism , Medicine, East Asian Traditional , Rhinitis, Allergic, Perennial/metabolism , Rhinitis, Allergic, Perennial/therapy , Toll-Like Receptor 4/genetics , Animals , Cell Line , Disease Models, Animal , Female , Lipopolysaccharides/pharmacology , Mice , Mice, Inbred C3H , RNA, Messenger/metabolism , Rhinitis, Allergic , Toll-Like Receptor 4/metabolism , Up-RegulationABSTRACT
Scorpion bite represents a significant and serious public health problem in certain regions of Brazil, as well as in other parts of the world. Inflammatory mediators are thought to be involved in the systemic and local immune response induced by Tityus serrulatus scorpion envenomation. The aim of this study was to evaluate the effect of extracts of Mimosa tenuiflora on model envenomation. In mice, the envenomation model is induced by Tityus serrulatus venom. Previous treatment of mice with fractions from M. tenuiflora was able to suppress the cell migration to the peritoneal cavity. The treatment of mice with M. tenuiflora extracts also decreased the levels of IL-6, IL-12, and IL-1ß. We concluded that the administration of the extract and fractions resulted in a reduction in cell migration and showed a reduction in the level of proinflammatory cytokines. This study demonstrates, for the first time, the anti-inflammatory effect of aqueous extract from the Mimosa tenuiflora plant on T. serrulatus venom.
Subject(s)
Gene Expression Regulation/drug effects , Inflammation/drug therapy , Plant Extracts/administration & dosage , Scorpion Venoms/toxicity , Animals , Brazil , Humans , Inflammation/chemically induced , Interleukin-12/biosynthesis , Interleukin-1beta/biosynthesis , Interleukin-6/biosynthesis , Mice , Mimosa/chemistry , Plant Extracts/chemistry , Scorpions/pathogenicityABSTRACT
Tumor-associated macrophages (TAMs) are preferentially M2 skewed and promote tumor growth, angiogenesis, invasion, and/or metastasis. In this study, we have analyzed the in vitro immunomodulatory potential of a non-toxic neem leaf glycoprotein (NLGP) in reprogramming Stage III supraglottic laryngeal tumor cell lysate (SLTCL) induced M2 TAMs to their classical anti-tumor shape (M1). Data generated from this study support that NLGP is effective in preventing the SLTCL induced generation (CD68(+)CD206(+)IL-10(high) to CD68(+)CD206(-)IL-10(low) TAMs) and functions (NO(low) to NO(high), MHC-I(low) to MHC-I(high), CD80(low) to CD80(high)) of pro-tumorous M2 macrophages, which in turn associated with sustained anti-tumor effector functions by promoting cytotoxic T cell activities and suppressing regulatory T cells. Furthermore, our data also suggest that NLGP prevents M2 skewness of TAMs by downregulating phosphorylation of targeted STAT3.
Subject(s)
Azadirachta/chemistry , Glycoproteins/pharmacology , Immune Evasion/drug effects , Immunologic Factors/pharmacology , Laryngeal Neoplasms/immunology , Macrophages/drug effects , Plant Leaves/chemistry , STAT3 Transcription Factor/metabolism , Cell Line, Tumor , Down-Regulation/drug effects , Humans , Interleukin-10/biosynthesis , Interleukin-12/biosynthesis , Macrophages/immunology , Nitric Oxide/biosynthesis , Nitric Oxide Synthase Type II/biosynthesis , Phosphorylation , Plant Extracts/pharmacology , RNA Interference , RNA, Small Interfering , STAT3 Transcription Factor/genetics , T-Lymphocytes, Cytotoxic/immunology , T-Lymphocytes, Regulatory/immunologyABSTRACT
Success of cancer vaccination is strongly hampered by immune suppression in the tumor microenvironment (TME). Interleukin (IL)-6 is particularly and highly produced by triple-negative breast cancer (TNBC) cells, and has been considered as an important contributor to immune suppression in the TME. Therefore, we hypothesized that IL-6 reduction may improve efficacy of vaccination against TNBC cancer through improved T-cell responses. To prove this hypothesis, we investigated the effect of curcumin, an inhibitor of IL-6 production, on vaccination of a highly attenuated Listeria monocytogenes (Listeria(at)), encoding tumor-associated antigens (TAA) Mage-b in a TNBC model 4T1. Two therapeutic vaccination strategies with Listeria(at)-Mage-b and curcumin were tested. The first immunization strategy involved all Listeria(at)-Mage-b vaccinations and curcumin after tumor development. As curcumin has been consumed all over the world, the second immunization strategy involved curcumin before and all therapeutic vaccinations with Listeria(at)-Mage-b after tumor development. Here, we demonstrate that curcumin significantly improves therapeutic efficacy of Listeria(at)-Mage-b with both immunization strategies particularly against metastases in a TNBC model (4T1). The combination therapy was slightly but significantly more effective against the metastases when curcumin was administered before compared to after tumor development. With curcumin before tumor development in the combination therapy, the production of IL-6 was significantly decreased and IL-12 increased by myeloid-derived suppressor cells (MDSC), in correlation with improved CD4 and CD8 T-cell responses in blood. Our study suggests that curcumin improves the efficacy of Listeria(at)-Mage-b vaccine against metastases in TNBC model 4T1 through reversal of tumor-induced immune suppression.
Subject(s)
Antineoplastic Agents/pharmacology , Bacterial Vaccines/immunology , Curcumin/pharmacology , Listeria monocytogenes/immunology , Mammary Neoplasms, Experimental , T-Lymphocyte Subsets/immunology , Triple Negative Breast Neoplasms/immunology , Triple Negative Breast Neoplasms/therapy , Animals , Antineoplastic Agents/administration & dosage , Apoptosis/drug effects , Apoptosis/immunology , Bacterial Vaccines/administration & dosage , Cancer Vaccines/administration & dosage , Cancer Vaccines/immunology , Cell Line, Tumor , Cell Proliferation/drug effects , Curcumin/administration & dosage , Disease Models, Animal , Female , Immunization , Interleukin-12/biosynthesis , Interleukin-6/biosynthesis , Mice , Myeloid Cells/drug effects , Myeloid Cells/immunology , Myeloid Cells/metabolism , Neoplasm Metastasis , T-Lymphocyte Subsets/metabolism , Triple Negative Breast Neoplasms/pathologyABSTRACT
Bunching onion [Allium fistulosum L. (Liliaceae)] secretes mucus in the cavities of its green leaves. The effects of the mucus, which is consumed as food, were examined. The mucus augmented the production of tumor necrosis factor (TNF)-α and monocyte chemotactic protein (MCP)-1 from RAW 264 cells and of interleukin (IL)-12 from J774.1 cells; however, extracts from green leaves and white sheaths did not. An oral administration of this mucus to mice augmented the immune functions of peritoneal cells by increasing TNF-α and IL-12 production and phagocytosis. It also augmented interferon (IFN)-γ production from spleen cells and natural killer (NK) activity. These results suggest that an oral administration of the A. fistulosum mucus can enhance natural immunity.
Subject(s)
Allium/chemistry , Immunologic Factors/administration & dosage , Immunologic Factors/pharmacology , Plant Extracts/administration & dosage , Plant Extracts/pharmacology , Administration, Oral , Animals , Cell Line , Interleukin-12/biosynthesis , Macrophages/drug effects , Macrophages/immunology , Macrophages/metabolism , Male , Mice , Mice, Inbred ICR , Plant Leaves/chemistry , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
Baicalin from Scutellaria baicalensis is a major flavonoid constituent found in the traditional Chinese medicinal herb Baikal skull cap. It has been widely used for the treatment of various diseases such as pneumonia, diarrhea, and hepatitis. Recent studies have demonstrated that baicalin possesses a wide range of pharmacological and biological activities, including anti-inflammatory, anti-microbial, anti-oxidant, and anti-tumor properties. Specifically, its anti-inflammatory activity has been estimated in various animal models of acute and chronic inflammation; however, its effects on dendritic cells (DCs) maturation and immuno-stimulatory activities are still unknown. In this study, we attempted to determine whether baicalin could influence DC surface molecule expression, antigen uptake capacity, cytokine production, and capacity to induce T-cell differentiation. Baicalin was shown to significantly suppress the expression of surface molecules CD80, CD86, major histocompatibility complex (MHC) class I, and MHC class II as well as the levels of interleukin-12 production in lipopolysaccharide stimulated DCs. Moreover, baicalin-treated DCs showed an impaired induction of the T helper type 1 immune response and a normal cell-mediated immune response. These findings provide important understanding of the immunopharmacological functions of baicalin and have ramifications for the development of therapeutic adjuvants for the treatment of DCs-related acute and chronic diseases.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Cell Differentiation/drug effects , Dendritic Cells/drug effects , Flavonoids/pharmacology , Scutellaria baicalensis , Th1 Cells/drug effects , Animals , B7-1 Antigen/biosynthesis , B7-2 Antigen/biosynthesis , Cells, Cultured , Dendritic Cells/cytology , Dendritic Cells/immunology , Dose-Response Relationship, Drug , Drugs, Chinese Herbal/pharmacology , Flavonoids/biosynthesis , Interleukin-12/biosynthesis , Lipopolysaccharides/pharmacology , Major Histocompatibility Complex/genetics , Male , Mice , Th1 Cells/cytology , Th1 Cells/immunologyABSTRACT
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.
Subject(s)
Genetic Therapy/methods , Granulocyte-Macrophage Colony-Stimulating Factor/genetics , Hyperthermia, Induced/methods , Interleukin-12/genetics , Adenoviridae/genetics , Animals , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/metabolism , Carcinoma, Non-Small-Cell Lung/therapy , Carcinoma, Non-Small-Cell Lung/virology , Cell Culture Techniques , Cell Line, Tumor , Granulocyte-Macrophage Colony-Stimulating Factor/biosynthesis , Granulocyte-Macrophage Colony-Stimulating Factor/pharmacokinetics , Humans , Interleukin-12/biosynthesis , Interleukin-12/pharmacokinetics , Liver Neoplasms/genetics , Liver Neoplasms/metabolism , Liver Neoplasms/therapy , Liver Neoplasms/virology , Lung Neoplasms/genetics , Lung Neoplasms/metabolism , Lung Neoplasms/therapy , Lung Neoplasms/virology , Mice , Mice, Inbred BALB C , Mice, Nude , Xenograft Model Antitumor AssaysABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: The fruits of Morus alba have been traditionally used as a tonic to enhance immune responses. MATERIALS AND METHODS: The macrophage activating constituents of Morus alba fruits were purified using various column chromatography techniques. The structures of isolated compounds were determined on the basis of spectroscopic data interpretation such as 1D and 2D NMR analysis. The macrophage activating activities of isolated compounds were evaluated by measuring the production of nitric oxide, TNF-α and IL-12 in RAW 264.7 cells. The phagocytic activity was also evaluated. RESULTS: Five pyrrole alkaloids, 5-(hydroxymethyl)-1H-pyrrole-2-carboxaldehyde (1), 2-formyl-1H-pyrrole-1-butanoic acid (2), 2-formyl-5-(hydroxymethyl)-1H-pyrrole-1-butanoic acid (3), 2-formyl-5-(methoxymethyl)-1H-pyrrole-1-butanoic acid (4) and Morrole A (5) were isolated from the fruits of Morus alba. Morrole A (5) is first reported in nature and other pyrrole alkaloids (1-4) are first reported from Morus species. Among the isolated compounds, compounds 3 and 4 significantly activated macrophage activity by the enhancement of nitric oxide, TNF-α and IL-12 production, and the stimulation of phagocytic activity in RAW 264.7 cells. CONCLUSION: Pyrrole alkaloids, including a new compound, were isolated from Morus alba fruits. These compounds activated macrophage activity in RAW 264.7 cells.
Subject(s)
Alkaloids/pharmacology , Fruit/chemistry , Macrophages/drug effects , Macrophages/metabolism , Phagocytosis/drug effects , Pyrroles/pharmacology , Alkaloids/analysis , Alkaloids/isolation & purification , Animals , Cell Line, Tumor , Cell Survival/drug effects , Interleukin-12/biosynthesis , Mice , Molecular Structure , Morus/chemistry , Nitric Oxide/biosynthesis , Plant Extracts/chemistry , Plant Extracts/pharmacology , Pyrroles/analysis , Pyrroles/isolation & purification , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
The aim of this study is to investigate phenotypic and functional modulation of murine dendritic cells (DCs) with use of purified Glycyrrhizin (GL). These impacts of GL on DCs both from bone marrow derived DCs and established DC cell 2.4 were assessed with conventional scanning electron microscopy (SEM), flow cytometry (FCM), transmission electron microscopy (TEM), cytochemistry assay, FITC-dextran, bio-assay and enzyme linked immunosorbent assay (ELISA). We found that the purified GL induced phenotypic maturation as evidenced by increased expression of CD86, CD40, CD80, CD83 and major histocompatibility complex II (MHC II). The functional tests showed the activity of acidic phosphatase (ACP) inside the DCs2.4 cells were down- regulated after treatment with GL (which occurs when phagocytosis of DCs2.4 cells were decreased). Finally, we proved that GL increased the production of IL-12, IL-10 and decreased the production of tumor necrosis factor alpha (TNF-α). These data indicated that GL could promote maturation of DCs and this adjuvant-like activity may have potential therapeutic value. It is therefore concluded that GL could exert positive modulation on murine DCs.
Subject(s)
Dendritic Cells/drug effects , Glycyrrhizic Acid/pharmacology , Acid Phosphatase/metabolism , Animals , Bone Marrow Cells , Cell Line , Cell Proliferation/drug effects , Cytoplasmic Vesicles/drug effects , Cytoplasmic Vesicles/ultrastructure , Dendritic Cells/ultrastructure , Enzyme Activation/physiology , Enzyme-Linked Immunosorbent Assay , Female , Flow Cytometry , Interleukin-10/biosynthesis , Interleukin-12/biosynthesis , Mice , Mice, Inbred C57BL , Microscopy, Electron, Scanning , Phagocytosis/drug effects , Phenotype , Pinocytosis/drug effects , Receptors, Tumor Necrosis Factor/biosynthesis , Receptors, Tumor Necrosis Factor/genetics , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
OBJECTIVES: We investigated whether Abatacept might reduce proinflammatory cytokine production by macrophages upon contact with cytokine activated T cells and/or stimulation with TLR ligands. METHODS: Macrophages and cytokine stimulated T cells (Tck) were added together in the presence of Abatacept or a control Ig, with or without TLR ligands. The production of cytokines was determined by luminex. RESULTS: Abatacept reduced Tck-induced production of TNFa by macrophages. Tck and TLR ligands synergistically induced the production of proinflammatory cytokines by macrophages, especially IL-12p70. The production of IL-12p70 coincided with the production of IFNg, which were both reduced in the presence of Abatacept. CONCLUSIONS: Tck induce the production of TNFa by macrophages and facilitate the highly increased production of proinflammatory cytokines in the presence of TLR ligands. Abatacept was shown to potently suppress these pathways suggesting that its role may extend beyond antigen specific T cell mediated effector function.
Subject(s)
Immunoconjugates/pharmacology , Immunosuppressive Agents/pharmacology , Macrophages/drug effects , T-Lymphocytes/immunology , Toll-Like Receptors/immunology , Abatacept , Cell Communication/immunology , Cells, Cultured , Coculture Techniques , Cytokines/biosynthesis , Cytokines/immunology , Drug Evaluation, Preclinical/methods , Humans , Inflammation Mediators/metabolism , Interleukin-12/biosynthesis , Ligands , Lymphocyte Activation/immunology , Macrophages/immunology , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
Acanthopanax koreanum is well known herb in traditional Korean, Chinese, and Japanese anti-inflammatory action without any adverse effects. In the current study, we investigated the inhibitory effects of isolated compounds 1-13 from the leaves of A. koreanum on the lipopolysaccharide-stimulated production of pro-inflammatory cytokines in bone marrow-derived dendritic cells. Of these lupane-type triterpenoids, 1 exhibited particularly high inhibitory effect on lipopolysaccharide-stimulated TNF-α, IL-6, and IL-12 production with the values ranging from 45.0 to 84.5% at a concentration of 50 µM. These results warrant further studies concerning the potential anti-inflammatory benefits of medicinal foods containing the leaves of A. koreanum.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Dendritic Cells/drug effects , Eleutherococcus/chemistry , Lipopolysaccharides/pharmacology , Triterpenes/pharmacology , Animals , Bone Marrow Cells/drug effects , Cells, Cultured , Cytokines/biosynthesis , Interleukin-12/biosynthesis , Interleukin-6/biosynthesis , Mice , Mice, Inbred C57BL , Plant Extracts/chemistry , Plant Extracts/pharmacology , Plant Leaves/chemistry , Tetrazolium Salts , Thiazoles , Triterpenes/chemistry , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
Radix Glycyrrhizae polysaccharide (GP) possesses multiple pharmacological activities. However, the effect of GP on CD4+CD25+ regulatory T (Treg) cells has not been elucidated. This study aimed to investigate the effects of GP on Treg cells and Th1/Th2 cytokines in H22 hepatocarcinoma tumor-bearing mice. The results demonstrated that GP inhibits tumor progression. In the lymph nodes of the tumor microenvironment and spleen, the proportion of Treg cells was significantly higher in the tumor-bearing mice. GP administration down-regulated the population of Treg cells (P < 0.01) and decreased lymph node Foxp3 and IL-10 mRNA expression (P < 0.01). In addition, GP treatment decreased IL-10 and TGF-ß level (P < 0.01) and increased IL-2 and IL-12p70 level in serum (P < 0.01). In conclusion, GP reduced the proportion of Treg cells and Foxp3 lowered expression in Treg cells, and up-regulated Th1/Th2 cytokine ratio in serum in the tumor bearing mice, which might partially cause the inhibition of tumor growth.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Carcinoma, Hepatocellular/drug therapy , Cytokines/metabolism , Drugs, Chinese Herbal/pharmacology , Glycyrrhiza , Polysaccharides/pharmacology , T-Lymphocytes, Regulatory/immunology , Th1 Cells/immunology , Animals , Carcinoma, Hepatocellular/immunology , Cell Proliferation/drug effects , Down-Regulation , Female , Forkhead Transcription Factors/biosynthesis , Interleukin-10/biosynthesis , Interleukin-12/biosynthesis , Liver Neoplasms/drug therapy , Liver Neoplasms/immunology , Mice , Mice, Inbred BALB C , Nitric Oxide/biosynthesis , Nitric Oxide Synthase/biosynthesis , Oxidative Stress/drug effects , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , T-Lymphocytes, Regulatory/drug effects , Th1 Cells/drug effects , Th1-Th2 Balance/drug effects , Th2 Cells/drug effects , Th2 Cells/immunology , Transforming Growth Factor beta/biosynthesisABSTRACT
We have previously demonstrated that in Ova-immunized mice the increase in intra-macrophage thiol pool induced by pro-GSH molecules modulates the Th1/Th2 balance in favour of a Th1-type immune response. We show now that the same molecules can support a Th1-type over Th2-type immunity against Tat, which is an early HIV-1 regulatory protein and a Th1 polarizing immunomodulator that is increasingly considered in new anti-HIV vaccination strategies. Our results indicate that Tat-immunized mice pre-treated with the C4 (n-butanoyl) derivative of reduced glutathione (GSH-C4) or a pro-drug of N-acetylcysteine (NAC) and beta-mercaptoethylamine (MEA) (I-152), have decreased levels of anti-Tat IgG1 as well as increased levels of anti-Tat IgG2a and IgG2b isotypes suggesting a Th1-type response. Moreover, Th1-(IFN-γ and IL-2) Ag-specific cellular responses were detected by ELISPOT assay in splenocytes of the same animals as well as an increase of IL-12 levels in the plasma. These findings suggest that the Th1 immune response to HIV-1 Tat could be further polarized by these molecules. These results together with those previously reported suggest that pro-GSH molecules could be used to modulate the immune response towards different antigens and may be further exploited for inducing specific Th1 immune responses against other HIV antigens as well as other intracellular pathogens in new Tat-based vaccination protocols.