ABSTRACT
Intensive research during the past decade has highlighted the impact of the regulatory function of the aryl hydrocarbon receptor (AhR) in immunity. In this study, we focused on the influence of AhR activation on the differentiation of murine bone marrow-derived myeloid precursor cells into mature macrophages. Our results show that the activation of AhR by subtoxic doses of the AhR ligand benzo(a)pyrene (BaP) impaired the proliferation of bone marrow cells (BMCs) whereas the proportion of resulting adherent cells was not affected. Flow cytometric analysis revealed that the number of mature bone marrow-derived macrophages (BMMs) was significantly decreased by AhR activation. However, expression of the murine macrophage marker F4/80, the major histocompatibility complex class II (MHC-II) and the Fcγ receptor I (FcγRI/CD64) were upregulated on BaP-exposed BMMs in an AhR-dependent manner. Analysis of cytokine secretion after BMM activation with heat-killed (hk) salmonellae showed that BaP exposure resulted in suppressed secretion of interleukin (IL)-1ß, IL-6 and the chemokine CXC motif ligand 1 (CXCL1). In contrast, the release of tumor necrosis factor (TNF)-α and IL-10 was increased following BaP exposure. In addition, the production of antimicrobial nitric oxide (NO) was increased AhR-dependently. Bacterial stimulation of BaP exposed BMMs also induced the expression of MHC-II and CD64, while the expression of F4/80 was dramatically decreased. In summary, this study demonstrates for the first time that sustained exposure over 6 days of bone marrow-derived myeloid precursors to subtoxic doses of BaP critically interferes with differentiation and activation of BMMs. We could convincingly show that AhR-induced gene regulation is crucial for homeostasis of pro- and anti-inflammatory cytokines during macrophage activation.
Subject(s)
Benzo(a)pyrene/pharmacology , Bone Marrow Cells/drug effects , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Macrophages/drug effects , Myeloid Progenitor Cells/drug effects , Receptors, Aryl Hydrocarbon/agonists , Animals , Antigens, Differentiation/metabolism , Biomarkers/metabolism , Cytokines/metabolism , Genes, MHC Class II/drug effects , Mice , Mice, Inbred C57BL , Nitric Oxide/metabolism , PhenotypeABSTRACT
Toxoplasma gondii is a ubiquitous apicomplexan parasite of mammals and birds and an important pathogen of humans. IFN-γ is the major mediator of host resistance against T. gondii but intriguingly, parasite-infected host cells including macrophages are severely impaired to respond to IFN-γ due to defective transcriptional activation of target genes. Here, we tested the possibility that the impaired responsiveness of T. gondii-infected macrophages to IFN-γ can be restored by inhibiting histone deacetylases (HDACs) using the class I-specific inhibitor MS-275. Treatment of RAW264.7 cells with MS-275 indeed increased MHC class II surface expression in infected and non-infected cells and largely abolished the inhibition of IFN-γ-regulated MHC class II expression exerted by T. gondii. Genome-wide transcriptome profiling revealed that MS-275 increased mean mRNA levels of IFN-γ-regulated genes particularly in non-infected macrophages. Transcript levels of 33% of IFN-γ secondary response genes but only those of a few primary response genes were also increased by MS-275 in T. gondii-infected cells. Importantly, the unresponsiveness of parasite-infected cells to IFN-γ was however not abolished by MS-275. Furthermore, MS-275 also up-regulated several anti-inflammatory cytokines or signaling molecules in T. gondii-infected macrophages. It additionally regulated expression of more than 2500 genes in non-infected macrophages expression of which was surprisingly counteracted by prior infection with T. gondii. FACS analysis and immunofluorescence microscopy revealed that MS-275 did not considerably diminish the number of parasite-positive cells or the intracellular replication in macrophages stimulated or not with IFN-γ. Thus, a supportive therapy using MS-275 appears inappropriate for treatment of toxoplasmosis.
Subject(s)
Benzamides/pharmacology , Gene Expression/drug effects , Histone Deacetylase Inhibitors/pharmacology , Interferon-gamma/genetics , Macrophages/drug effects , Pyridines/pharmacology , Toxoplasma/drug effects , Animals , Flow Cytometry , Genes, MHC Class II/drug effects , Histone Deacetylases/drug effects , Interferon-gamma/physiology , Macrophages/immunology , Macrophages/parasitology , Mice , Microscopy, Fluorescence , RAW 264.7 Cells , RNA, Protozoan/chemistry , RNA, Protozoan/isolation & purification , Toxoplasma/genetics , Toxoplasma/immunologyABSTRACT
Th2 immunity plays important roles in both protective and allergic responses. Nevertheless, the nature of antigen-presenting cells responsible for Th2 cell differentiation remains ill-defined compared with the nature of the cells responsible for Th1 and Th17 cell differentiation. Basophils have attracted attention as a producer of Th2-inducing cytokine IL-4, whereas their MHC class II (MHC-II) expression and function as antigen-presenting cells are matters of considerable controversy. Here we revisited the MHC-II expression on basophils and explored its functional relevance in Th2 cell differentiation. Basophils generated in vitro from bone marrow cells in culture with IL-3 plus GM-CSF displayed MHC-II on the cell surface, whereas those generated in culture with IL-3 alone did not. Of note, these MHC-II-expressing basophils showed little or no transcription of the corresponding MHC-II gene. The GM-CSF addition to culture expanded dendritic cells (DCs) other than basophils. Coculture of basophils and DCs revealed that basophils acquired peptide-MHC-II complexes from DCs via cell contact-dependent trogocytosis. The acquired complexes, together with CD86, enabled basophils to stimulate peptide-specific T cells, leading to their proliferation and IL-4 production, indicating that basophils can function as antigen-presenting cells for Th2 cell differentiation. Transfer of MHC-II from DCs to basophils was also detected in draining lymph nodes of mice with atopic dermatitis-like skin inflammation. Thus, the present study defined the mechanism by which basophils display MHC-II on the cell surface and appears to reconcile some discrepancies observed in previous studies.
Subject(s)
Antigen Presentation/immunology , Antigen-Presenting Cells/immunology , Basophils/immunology , Dendritic Cells/immunology , Histocompatibility Antigens Class II/immunology , Immunological Synapses/immunology , Amino Acid Sequence , Animals , Calcitriol/analogs & derivatives , Calcitriol/toxicity , Cells, Cultured , Coculture Techniques , Cytokines/pharmacology , Dermatitis, Contact/immunology , Dermatitis, Contact/pathology , Genes, MHC Class II/drug effects , Histocompatibility Antigens Class II/biosynthesis , Histocompatibility Antigens Class II/genetics , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Transgenic , Peptide Fragments/immunology , Specific Pathogen-Free Organisms , Th2 Cells/immunology , Tryptases/geneticsABSTRACT
To stimulate a productive T cell response, dendritic cells (DC) must undergo maturation characterized by heightened cell surface expression of MHC and costimulatory molecules as well as cytokine production. Conversely, the inhibition of DC maturation is a central mechanism of immune tolerance. The control of the DC maturation process relies on the integration of several cellular stimulatory or inhibitory signals. The soluble factors and their receptors controlling this central aspect of DC biology are incompletely characterized. We show that murine bone marrow-derived DC (BMDC) maturation induced by LPS, as opposed to polyinosinic:polycytidylic acid or cytosine-phosphate-guanine, is robustly inhibited by vascular endothelial growth factor (VEGF), a previously identified immunosuppressive cytokine. Using BMDC from wild type and conditional knockout mice, we show that neuropilin-1 (NRP-1), a known receptor of VEGF, is necessary to suppress LPS-dependent BMDC maturation. The absence of NRP-1 had no ostensible effects on the biology of BMDC in the absence of VEGF. However, NRP-1-deficient BMDC remained completely insensitive to the VEGF-dependent inhibition of BMDC maturation in culture. In the presence of VEGF, NRP-1 directly interacted with the LPS receptor TLR4 and suppressed downstream signaling through ERK and NF-κß, resulting in a sharp inhibition of MHC class II and costimulatory molecules (CD40, CD86) expression as well as proinflammatory cytokine production. Consequently, we identify NRP-1 as a target to optimize DC maturation within environments that are rich in VEGF, such as tumors.
Subject(s)
Dendritic Cells/drug effects , Dendritic Cells/physiology , Neuropilin-1/immunology , Neuropilin-1/metabolism , Vascular Endothelial Growth Factor A/metabolism , Animals , B7-2 Antigen/drug effects , B7-2 Antigen/genetics , Bone Marrow Cells/immunology , Bone Marrow Cells/physiology , CD40 Antigens/drug effects , CD40 Antigens/genetics , Cell Differentiation , Cells, Cultured , Cytokines/biosynthesis , Cytokines/drug effects , Cytokines/genetics , Dendritic Cells/immunology , Genes, MHC Class II/drug effects , Genes, MHC Class II/genetics , Immune Tolerance/drug effects , Lipopolysaccharides/immunology , MAP Kinase Signaling System/physiology , Mice , NF-kappa B p50 Subunit/physiology , Neuropilin-1/deficiency , Poly I-C/pharmacology , Signal Transduction/immunology , Signal Transduction/physiology , Toll-Like Receptor 4/metabolism , Vascular Endothelial Growth Factor A/pharmacologyABSTRACT
ß-glucan is a prebiotic well known for its beneficial outcomes on sea cucumber health through modifying the host intestinal microbiota. High-throughput sequencing techniques provide an opportunity for the identification and characterization of microbes. In this study, we investigated the intestinal microbial community composition, interaction among species, and intestinal immune genes in sea cucumber fed with diet supplemented with or without ß-glucan supplementation. The results show that the intestinal dominant classes in the control group are Flavobacteriia, Gammaproteobacteria, and Alphaproteobacteria, whereas Alphaproteobacteria, Flavobacteriia, and Verrucomicrobiae are enriched in the ß-glucan group. Dietary ß-glucan supplementation promoted the proliferation of the family Rhodobacteraceae of the Alphaproteobacteria class and the family Verrucomicrobiaceae of the Verrucomicrobiae class and reduced the relative abundance of the family Flavobacteriaceae of Flavobacteria class. The ecological network analysis suggests that dietary ß-glucan supplementation can alter the network interactions among different microbial functional groups by changing the microbial community composition and topological roles of the OTUs in the ecological network. Dietary ß-glucan supplementation has a positive impact on immune responses of the intestine of sea cucumber by activating NF-κB signaling pathway, probably through modulating the balance of intestinal microbiota.
Subject(s)
Genes, MHC Class II/drug effects , Intestines/immunology , Intestines/microbiology , Stichopus , beta-Glucans/pharmacology , Animal Feed , Animals , Dietary Supplements , Gene Expression Regulation/drug effects , Intestines/drug effects , Microbiota/drug effects , Rhodobacteraceae/drug effectsABSTRACT
BACKGROUND: A close association between periodontitis and diabetes has been demonstrated in human cross-sectional studies, but an exact relationship between periodontitis and prediabetes has not been established. Previous studies using animal model systems consistently have shown that hyperinsulinemia occurs in animals with periodontitis compared to animals with healthy periodontium (while maintaining normoglycemia). Because bacterial lipopolysaccharide (LPS) plays an important role in the pathogenesis of periodontitis, we hypothesized that LPS may stimulate insulin secretion through a direct effect on ß cell function. To test this hypothesis, pancreatic ß cell line MIN6 cells were used to determine the effect of Porphyromonas gingivalis (Pg) LPS on insulin secretion. Furthermore, expression of genes altered by Pg LPS in innate immunity and insulin-signaling pathways was determined. METHODS: MIN6 cells were grown in medium with glucose concentration of normoglycemia (5.5 mM). Pg LPS was added to each well at final concentrations of 50, 200, and 500 ng/mL. Insulin secretion was measured using enzyme-linked immunosorbent assay. Gene expression levels altered by Pg LPS were determined by polymerase chain reaction (PCR) array for mouse innate and adaptive immunity response and mouse insulin-signaling pathways, and results were confirmed for specific genes of interest by quantitative PCR. RESULTS: Pg LPS stimulated insulin secretion in the normoglycemic condition by ≈1.5- to 3.0-fold depending on the concentration of LPS. Pg LPS treatment altered the expression of several genes involved in innate and adaptive immune response and insulin-signaling pathway. Pg LPS upregulated the expression of the immune response-related genes cluster of differentiation 8a (Cd8a), Cd14, and intercellular adhesion molecule-1 (Icam1) by about two-fold. LPS also increased the expression of two insulin signaling-related genes, glucose-6-phosphatase catalytic subunit (G6pc) and insulin-like 3 (Insl3), by three- to four-fold. CONCLUSIONS: We have demonstrated for the first time that Pg LPS stimulates insulin secretion by pancreatic ß cell line MIN cells. Pg LPS may have significant implications on the development of ß cell compensation and insulin resistance in prediabetes in individuals with periodontitis.
Subject(s)
Insulin-Secreting Cells/drug effects , Insulin/metabolism , Lipopolysaccharides/pharmacology , Porphyromonas gingivalis/physiology , Adaptive Immunity/drug effects , Adaptive Immunity/genetics , Animals , CD8 Antigens/drug effects , Cell Culture Techniques , Cell Line , Genes, MHC Class II/drug effects , Glucose/pharmacology , Glucose-6-Phosphatase/drug effects , Humans , Immunity, Innate/drug effects , Immunity, Innate/genetics , Insulin Secretion , Insulin-Secreting Cells/metabolism , Intercellular Adhesion Molecule-1/drug effects , Lipopolysaccharide Receptors/drug effects , Lipopolysaccharides/immunology , Mice , Porphyromonas gingivalis/immunology , Proteins/drug effects , Signal Transduction/drug effects , Signal Transduction/genetics , Up-RegulationABSTRACT
Pancreatic cancer is characterized as a type of gastrointestinal tumor with a poor prognosis and high degree of malignancy. CIITA gene was found highly methylated in pancreatic carcinoma cell line PANC-1 and responsible for the low expression of MHC-II that may lead to immune evasion. Here, we tried to prepare pancreatic cancer vaccine with PANC-1 cells via epigenetic modification to enhance the MHC-II expression. Then the vaccine was injected into C57BL/6J mice and the effect was examined. Our study found that the vaccine could promote the proliferation of antigen-specific T cells, enhance the killing activity of cytotoxic lymphocytes (CTL), promote Th1-type cells mediated secretion of cytokines IFN-gamma and IL-2 while inhibiting Th2-type cells mediated secretion of IL-4, and inhibit the secretion of TGF-beta. Generally, the epigenetically modified vaccine could enhance the body's anti-tumor immune response, providing feasibility research on cancer vaccine for therapy of pancreatic cancer.
Subject(s)
Cancer Vaccines/genetics , DNA Methylation/genetics , Epigenesis, Genetic/genetics , Pancreatic Neoplasms/immunology , Animals , Cancer Vaccines/immunology , Cell Death/drug effects , Cell Death/genetics , Cell Line, Tumor , Cell Proliferation/drug effects , Cells, Cultured , Cytokines/analysis , Cytokines/metabolism , DNA (Cytosine-5-)-Methyltransferases/antagonists & inhibitors , DNA Methylation/drug effects , Enzyme Inhibitors/pharmacology , Epigenesis, Genetic/drug effects , Genes, MHC Class II/drug effects , Genes, MHC Class II/genetics , Histone Deacetylase Inhibitors/pharmacology , Humans , Mice , Mice, Inbred C57BL , Nuclear Proteins/drug effects , Nuclear Proteins/genetics , Pancreatic Neoplasms/genetics , Spleen/cytology , Th1 Cells/metabolism , Th2 Cells/metabolism , Trans-Activators/drug effects , Trans-Activators/geneticsABSTRACT
BACKGROUND: The aim of this study was to construct a novel prediction model for the pathological complete response (pCR) to neoadjuvant chemotherapy (NAC) using immune-related gene expression data. PATIENTS AND METHODS: DNA microarray data were used to perform a gene expression analysis of tumor samples obtained before NAC from 117 primary breast cancer patients. The samples were randomly divided into the training (n = 58) and the internal validation (n = 59) sets that were used to construct the prediction model for pCR. The model was further validated using an external validation set consisting of 901 patients treated with NAC from six public datasets. RESULTS: The training set was used to construct an immune-related 23-gene signature for NAC (IRSN-23) that is capable of classifying the patients as either genomically predicted responders (Gp-R) or non-responders (Gp-NR). IRSN-23 was first validated using an internal validation set, and the results showed that the pCR rate for Gp-R was significantly higher than that obtained for Gp-NR (38 versus 0%, P = 1.04E-04). The model was then tested using an external validation set, and this analysis showed that the pCR rate for Gp-R was also significantly higher (40 versus 11%, P = 4.98E-23). IRSN-23 predicted pCR regardless of the intrinsic subtypes (PAM50) and chemotherapeutic regimens, and a multivariate analysis showed that IRSN-23 was the most important predictor of pCR (odds ratio = 4.6; 95% confidence interval = 2.7-7.7; P = 8.25E-09). CONCLUSION: The novel prediction model (IRSN-23) constructed with immune-related genes can predict pCR independently of the intrinsic subtypes and chemotherapeutic regimens.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Breast Neoplasms/genetics , Transcriptome/immunology , Breast Neoplasms/drug therapy , Breast Neoplasms/immunology , Chemotherapy, Adjuvant , Cyclophosphamide/administration & dosage , Epirubicin/administration & dosage , Female , Fluorouracil/administration & dosage , Genes, MHC Class II/drug effects , Humans , Middle Aged , Models, Biological , Multivariate Analysis , Neoadjuvant Therapy , Paclitaxel/administration & dosage , Prognosis , Retrospective Studies , Treatment OutcomeABSTRACT
BACKGROUND: Mesenchymal stem cells (MSCs) are multipotent stromal cells that can differentiate into a variety of cell types. The MSCs can be activated and mobilized if needed. AIM: This study aimed to investigate the response mechanism of MSCs under Dexamethasone (Dex) treatment by combining MSCs microarray and bioinformatics methods. MATERIALS AND METHODS: We downloaded the gene expression profile of rat's MSCs challenge with or without Dex (GSE3339) from Gene Expression Omnibus database, including 2 Dex treated samples and 3 untreated samples. The differentially expressed genes (DEGs) were identified by packages in R language. Then, Gestalt (Genomic Sequence Total Analysis and Lookup Tool) and EASE (Expression Analysis Systematic Explorer) to were employed to obtain the molecular events of MSCs under Dex treatment. RESULTS: A total of 17 genes were identified as DEGs between untreated and treated samples, and they were significant enriched in immune response and cell differentiation. The C3 gene was the common candidate gene selected from two different algorithms, and 24 conserved sites were identified in the 3'UTR of C3 gene. CONCLUSIONS: Genes associated with immune response and cell differentiation were dysregulated in MSCs under Dex.
Subject(s)
Dexamethasone/pharmacology , Gene Expression Regulation/drug effects , Glucocorticoids/pharmacology , Mesenchymal Stem Cells/drug effects , Algorithms , Animals , Cell Differentiation/drug effects , Cell Differentiation/genetics , Computational Biology , Databases, Genetic , Genes, MHC Class II/drug effects , Mesenchymal Stem Cells/metabolism , Oligonucleotide Array Sequence Analysis , RatsABSTRACT
Since the inflammatory response and oxidative stress are involved in the stroke cascade, we evaluated here the effects of Phycocyanobilin (PCB, the C-Phycocyanin linked tetrapyrrole) on PC12 cell survival, the gene expression and the oxidative status of hypoperfused rat brain. After the permanent bilateral common carotid arteries occlusion (BCCAo), the animals were treated with saline or PCB, taking samples 24h post-surgery. Global gene expression was analyzed with GeneChip Rat Gene ST 1.1 from Affymetrix; the expression of particular genes was assessed by the Fast SYBR Green RT-PCR Master Mix and Bioplex methods; and redox markers (MDA, PP, CAT, SOD) were evaluated spectrophotometrically. The PCB treatment prevented the H2O2 and glutamate induced PC12 cell injury assessed by the MTT assay, and modulated 190 genes (93 up- and 97 down-regulated) associated to several immunological and inflammatory processes in BCCAo rats. Furthermore, PCB positively modulated 19 genes mostly related to a detrimental pro-inflammatory environment and counteracted the oxidative imbalance in the treated BCCAo animals. Our results support the view of an effective influence of PCB on major inflammatory mediators in acute cerebral hypoperfusion. These results suggest that PCB has a potential to be a treatment for ischemic stroke for which further studies are needed.
Subject(s)
Cell Survival/drug effects , Cerebrovascular Disorders/drug therapy , Genes, MHC Class II/drug effects , Inflammation/genetics , Oxidative Stress/drug effects , Phycobilins/pharmacology , Phycocyanin/pharmacology , Animals , Biomarkers/metabolism , Brain Chemistry/drug effects , Brain Chemistry/genetics , Cerebrovascular Disorders/physiopathology , Coloring Agents , Cytokines/biosynthesis , Glutamic Acid/metabolism , Hydrogen Peroxide/pharmacology , Male , Microarray Analysis , Oxidation-Reduction , PC12 Cells , Phycobilins/isolation & purification , Phycocyanin/isolation & purification , Rats , Rats, Wistar , Real-Time Polymerase Chain Reaction , Spirulina/chemistry , Tetrazolium Salts , Thiazoles , Vascular Endothelial Growth Factor A/metabolismABSTRACT
Treatment of neonatal mice with the phytoestrogen genistein (50 mg/kg/day) results in complete female infertility caused in part by preimplantation embryo loss in the oviduct between Days 2 and 3 of pregnancy. We previously demonstrated that oviducts of genistein-treated mice are "posteriorized" as compared to control mouse oviducts because they express numerous genes normally restricted to posterior regions of the female reproductive tract (FRT), the cervix and vagina. We report here that neonatal genistein treatment resulted in substantial changes in oviduct expression of genes important for the FRT mucosal immune response, including immunoglobulins, antimicrobials, and chemokines. Some of the altered immune response genes were chronically altered beginning at the time of neonatal genistein treatment, indicating that these alterations were a result of the posteriorization phenotype. Other alterations in oviduct gene expression were observed only in early pregnancy, immediately after the FRT was exposed to inflammatory or antigenic stimuli from ovulation and mating. The oviduct changes affected development of the surviving embryos by increasing the rate of cleavage and decreasing the trophectoderm-to-inner cell mass cell ratio at the blastocyst stage. We conclude that both altered immune responses to pregnancy and deficits in oviduct support for preimplantation embryo development in the neonatal genistein model are likely to contribute to infertility phenotype.
Subject(s)
Embryonic Development/drug effects , Genistein/toxicity , Immunity, Mucosal/drug effects , Oviducts/drug effects , Oviducts/immunology , Phytoestrogens/toxicity , Animals , Animals, Newborn , Embryonic Development/genetics , Embryonic Development/immunology , Embryonic Development/physiology , Female , Gene Expression Regulation, Developmental/drug effects , Genes, MHC Class II/drug effects , Genistein/administration & dosage , Immunity, Mucosal/genetics , Infertility, Female/chemically induced , Infertility, Female/genetics , Infertility, Female/immunology , Infertility, Female/metabolism , Inflammation Mediators/metabolism , Male , Mice , Oviducts/metabolism , Oviducts/pathology , Phytoestrogens/administration & dosage , PregnancyABSTRACT
Methionine enkephalin (MENK), the endogenous neuropeptide, is known to exert direct effects on the neuroendocrine and the immune systems and participates in regulation of various functions of cells related to both the innate and adaptive immune systems. Dendritic cells (DCs) play important role in initiating and regulating T cell responses. The aim of this work is to investigate the effects of MENK on differentiation, maturation, and function of DCs derived from murine bone marrow progenitors (BM-derived DCs). Our result showed that MENK could induce BM-derived DCs to polarize predominantly to mDC subtype, rather than pDC both in vivo and in vitro, and this was in favor of Th1 response. BM-derived DCs, after treatment with MENK, up-regulated the expressions of MHC class II and key costimulatory molecules. Result by RT-PCR showed MENK could increase expressions of delta and kappa receptors on BM-derived DCs. Also MENK promoted BM-derived DCs to secret higher levels of proinflammatory cytokines of IL-12p70, TNF-α. Furthermore, differentiated BM-derived DCs treated with MENK displayed higher activity to induce allogeneic T cell proliferation and MENK also inhibited tumor growth in vivo and induced apoptosis of tumor cells in vitro. Thus, it is concluded that MENK could be an effective inducer of BM-derived DCs and might be a new therapeutic agent for cancer, as well as other immune handicapped disease. Also we may consider MENK as a potential adjuvant in vaccine preparation.
Subject(s)
Bone Marrow Cells/drug effects , Cell Differentiation/drug effects , Dendritic Cells/drug effects , Enkephalin, Methionine/pharmacology , Neurotransmitter Agents/pharmacology , Stem Cells/drug effects , Animals , Apoptosis/drug effects , Cell Line, Tumor , Cytokines/metabolism , Female , Genes, MHC Class II/drug effects , Humans , Lymphocyte Activation/drug effects , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Receptors, Opioid, delta/biosynthesis , Receptors, Opioid, kappa/biosynthesis , Up-Regulation/drug effectsABSTRACT
There is a considerable inter-individual variation in response to statin therapy and one third of patients do not meet their treatment goals. We aimed to identify differentially expressed genes that might be involved in the effects of statin treatment and to suggest potential markers to guide statin therapy. Forty-six healthy Korean subjects received atorvastatin; their whole-genome expression profiles in peripheral blood were analyzed before and after atorvastatin administration in relation with changes in lipid profiles. The expression patterns of the differentially expressed genes were also compared with the data of familial hypercholesterolemia (FH) patients and controls. Pairwise comparison analyses revealed differentially expressed genes involved in diverse biological processes and molecular functions related with immune responses. Atorvastain mainly affected antigen binding, immune or inflammatory response including interleukin pathways. Similar expression patterns of the genes were observed in patients with FH and controls. The Charcol-Leyden crystal (CLC), CCR2, CX3CR1, LRRN3, FOS, LDLR, HLA-DRB1, ERMN, and TCN1 genes were significantly associated with cholesterol levels or statin response. Interestingly, the CLC gene, which was significantly altered by atorvastatin administration and differentially expressed between FH patients and controls, showed much bigger change in high-responsive group than in low-responsive group. We identified differentially expressed genes that might be involved in mechanisms underlying the known pleiotropic effects of atorvastatin, baseline cholesterol levels, and drug response. Our findings suggest CLC as a new candidate marker for statin response, and further validation is needed.
Subject(s)
Cholesterol/blood , Gene Expression/drug effects , Heptanoic Acids/administration & dosage , Hypercholesterolemia/blood , Pyrroles/administration & dosage , Adult , Atorvastatin , Biomarkers, Pharmacological/blood , Gene Expression Profiling/methods , Genes, MHC Class II/drug effects , Genome, Human , Humans , Hypercholesterolemia/genetics , MaleABSTRACT
There are a large number of interactions at molecular and cellular levels between the plant polysaccharides and immune system. Plant polysaccharides present an interesting effects as immunomodulators, particularly in the induction of the cells both in innate and adaptive immune systems. Activation of DCs could improve antitumoral responses usually diminished in cancer patients, and natural adjuvants provide a possibility of inducing this activation. ABP is a purified polysaccharide isolated from Achyranthes bidentata, a traditional Chinese medicine (TCM). The aim of this study is to investigate modulation of phenotypic and functional maturation of murine DCs by ABP. Both phenotypic and functional activities were assessed with use of conventional scanning electronic microscopy (SEM) for the morphology of the DC, transmitted electron microscopy (TEM) for intracellular lysosomes inside the DC, cellular immunohistochemistry for phagocytosis by the DCs, flow cytometry (FCM) for the changes in key surface molecules, bio-assay for the activity of acidic phosphatases (ACP), and ELISA for the production of pro-inflammatory cytokine IL-12. In fact, we found that purified ABP induced phenotypic maturation revealed by increased expression of CD86, CD40, and MHC II. Functional experiments showed the down-regulation of ACP inside DCs (which occurs when phagocytosis of DCs is decreased, and antigen presentation increased with maturation). Finally, ABP increased the production of IL-12. These data reveal that ABP promotes effective activation of murine DCs. This adjuvant-like activity may have therapeutic applications in clinical settings where immune responses need boosting. It is therefore concluded that ABP can exert positive modulation to murine DCs.
Subject(s)
Achyranthes/chemistry , Dendritic Cells/cytology , Dendritic Cells/drug effects , Polysaccharides/pharmacology , Achyranthes/immunology , Acid Phosphatase/metabolism , Adjuvants, Immunologic/pharmacology , Animals , Antigen Presentation/drug effects , Antigen Presentation/immunology , B7-2 Antigen/biosynthesis , CD40 Antigens/biosynthesis , Cell Differentiation/drug effects , Cell Differentiation/immunology , Cell Line , Dendritic Cells/immunology , Dendritic Cells/metabolism , Down-Regulation , Genes, MHC Class II/drug effects , Genes, MHC Class II/genetics , Interleukin-12/biosynthesis , Lysosomes/ultrastructure , Mice , Phagocytosis/immunology , Phenotype , Plant Extracts/immunology , Plant Extracts/isolation & purification , Plant Extracts/pharmacology , Polysaccharides/immunology , Polysaccharides/isolation & purificationABSTRACT
Genetic regulation of autoimmune neuroinflammation is a well known phenomenon, but genetic influences on inflammation following traumatic nerve injuries have received little attention. In this study we examined the inflammatory response in a rat traumatic brain injury (TBI) model, with a particular focus on major histocompatibility class II (MHC II) presentation, in two inbred rat strains that have been extensively characterized in experimental autoimmune encephalomyelitis (EAE); DA and PVG. In addition, MHC and Vra4 congenic strains on these backgrounds were studied to give information on MHC and non-MHC gene contribution. Thus, allelic differences in Vra4, harboring the Ciita gene, was found to regulate expression of the invariant chain at the mRNA level, with a much smaller effect exerted by the MHC locus itself. Notably, however, at the protein level the MHC congenic PVG-RT1(av1) strain displayed much stronger MHCII(+) presentation, as shown both by immunolabeling and flow cytometry, than the PVG strain, dwarfing the effect of Ciita. The PVG-RT1(av1) strain had significantly more T-cell influx than both DA and PVG, suggesting regulation both by MHC and non-MHC genes. Finally, in terms of outcome, the EAE susceptible DA strain displayed a significantly smaller resulting lesion volume than the resistant PVG-RT1(av1) strain. These results provide additional support for a role of adaptive immune response after neurotrauma and demonstrate that outcome is significantly affected by host genetic factors.
Subject(s)
Brain Injuries/immunology , Inflammation/immunology , Major Histocompatibility Complex/immunology , T-Lymphocytes/immunology , Animals , Brain/drug effects , Brain/immunology , Brain/pathology , Brain Injuries/pathology , Flow Cytometry , Genes, MHC Class II/drug effects , Genes, MHC Class II/immunology , Genes, MHC Class II/physiology , Inflammation/physiopathology , Major Histocompatibility Complex/drug effects , Major Histocompatibility Complex/physiology , Male , Myelin Basic Protein/pharmacology , Peptide Fragments/pharmacology , Rats , Rats, Inbred Strains , Reverse Transcriptase Polymerase Chain Reaction , T-Lymphocytes/physiologyABSTRACT
Fine particulate air pollutants, mainly their organic fraction, have been demonstrated to be associated with cardiovascular and respiratory health problems. Puerto Rico has been reported to have the highest prevalence of pulmonary diseases (e.g., asthma) in the United States. The aim of this study was to assess, for the first time, the immunological response of human bronchial epithelial cells (BEAS-2B) to organic extracts isolated from airborne particulate matter (PM(2.5)) in Puerto Rico. Organic extracts from PM(2.5) collected throughout an 8-month period (2000-2001) were pooled (composite) in order to perform chemical analysis and biological activity testing. BEAS-2B cells were exposed to PM(2.5) organic extract to assess cytotoxicity, levels of cytokines and relative gene expression of MHC-II, hPXR and CYP3A5. Our findings show that organic PM(2.5) consist of toxic as well as bioactive components that can regulate the secretion of cytokines in BEAS-2B, which could modulate inflammatory response in the lung. Trace element analyses confirmed the presence of metals in organic extracts highlighting the relative high abundance of Cu and Zn in polar organic extracts. Polar organic extracts exhibited dose-dependant toxicity and were found to significantly induce the release of interleukin 6 (IL-6), IL-1beta and IL-7 while significantly inhibiting the secretion of IL-8, G-CSF and MCP-1. Moreover, MHC-II transcriptional activity was up-regulated after 24 h of exposure, whereas PXR and CYP3A5 were down-regulated. This research provides a new insight into the effects of PM(2.5) organic fractions on specific effectors and their possible role in the development of respiratory inflammatory diseases in Puerto Rico.
Subject(s)
Air Pollutants, Occupational/toxicity , Biomarkers/analysis , Bronchi/pathology , Epithelial Cells/immunology , Epithelial Cells/pathology , Particulate Matter/toxicity , Air Pollutants, Occupational/analysis , Bronchi/cytology , Bronchi/immunology , Cell Line , Cytochrome P-450 CYP3A/biosynthesis , Cytokines/analysis , Environmental Monitoring , Genes, MHC Class II/drug effects , HLA-DR2 Antigen/biosynthesis , Humans , Particulate Matter/analysis , Pregnane X Receptor , Puerto Rico , RNA, Messenger/biosynthesis , RNA, Messenger/isolation & purification , Receptors, Steroid/biosynthesis , Reverse Transcriptase Polymerase Chain Reaction , Suppression, Genetic/drug effects , Trace Elements/toxicity , Transcriptional ActivationABSTRACT
Nivalenol (NIV) and Deoxynivalenol (DON), mycotoxins of the trichothecene family are considered very common food contaminants. In this work, we investigated whether the immunotoxic effects ascribed to these trichothecenes may be mediated by perturbations in the activity of dendritic cells (DCs). Murine bone marrow-derived DCs were used to evaluate the effects of NIV and DON on the LPS-induced maturation process. We found that the expression of the class II MHC and of the accessory CD11c molecules, but not of the costimulatory CD86 marker, was down-regulated by NIV and DON exposure in LPS-treated DCs, as well as nitric oxide (NO) production. Interestingly, NIV, but not DON, induced DC necrosis. Moreover, the analysis of the cytokine pattern showed that IL-12 and IL-10 expressions induced by LPS exposure were suppressed by both trichothecenes in a dose-dependent fashion. On the other hand, the secretion of the proinflammatory cytokine TNF-alpha was increased as a direct consequence of DON and NIV exposure. Taken together, our data indicated that the immunotoxicity of NIV and DON was related to the capacity of both trichothecenes to interfere with phenotypic and functional features of maturing DCs.
Subject(s)
Cell Differentiation/drug effects , Dendritic Cells/drug effects , Mycotoxins/toxicity , Trichothecenes/toxicity , Animals , Bone Marrow Cells , CD11c Antigen/metabolism , Cell Survival/drug effects , Cells, Cultured , Cytokines/metabolism , Dendritic Cells/cytology , Dendritic Cells/metabolism , Dose-Response Relationship, Drug , Flow Cytometry , Genes, MHC Class II/drug effects , Lipopolysaccharides/pharmacology , Mice , Mice, Inbred BALB C , Necrosis/chemically induced , Nitric Oxide/metabolismABSTRACT
Major histocompatibility complex (MHC) class II molecules play crucial roles in adaptive immune response and antigen presentation. Owing to enlargement of capsaicin's availability as an anti-inflammatory agent in medical therapeutics, we investigated the new effects of capsaicin that are related to adaptive immune response in terms of MHC class II expression in murine primary cultured macrophages. Capsaicin (0.1-10microM) reduced the expression of MHC class II mRNA levels in cultured peritoneal macrophages upon treatment with interferon (IFN)-gamma (100units/ml). In agreement with this, treatment of the cells with capsaicin also inhibited MHC class II transactivator (CIITA) mRNA expression induced by IFN-gamma in a dose-dependent manner. In contrast, production of nitric oxide, which has the ability to reduce MHC class II expression, was not enhanced but rather suppressed by capsaicin treatment in IFN-gamma-stimulated macrophages. These findings suggest that capsaicin suppresses expression of MHC class II via downregulation of CIITA transcription but not through the mediation of nitric oxide production by macrophages. These new immunopharmacological roles of capsaicin in specific transcription regulation of genes involved in antigen presentation of macrophages could be useful for the treatment of adaptive immune-mediated disorders.
Subject(s)
Capsaicin/pharmacology , Macrophages, Peritoneal/drug effects , Nuclear Proteins/biosynthesis , Trans-Activators/biosynthesis , Adaptive Immunity , Animals , Antigen Presentation , Cells, Cultured , Dose-Response Relationship, Drug , Drug Antagonism , Genes, MHC Class II/drug effects , Genes, MHC Class II/genetics , Interferon-gamma/pharmacology , Macrophages, Peritoneal/immunology , Macrophages, Peritoneal/metabolism , Macrophages, Peritoneal/pathology , Male , Mice , Mice, Inbred Strains , Nitric Oxide/metabolism , Nuclear Proteins/genetics , Trans-Activators/genetics , Transcription, Genetic/drug effectsABSTRACT
AIM: (5R)-5-hydroxytriptolide (LLDT-8) displayed immunosuppressive activities both in vitro and in autoimmune disease models. Here, we aim to further clarify the effect of LLDT-8 on the immune responses of human peripheral blood mononuclear cells (PBMC). METHOD: Cell proliferation of human PBMC from healthy donors was evaluated by [3H]-thymidine uptake. NK cell cytotoxicity was assayed using K562 cells in a [3H] lysis assay. Cytokine production was determined by enzyme-linked immunosorbent assay. The expression of cell surface molecules was detected with flow cytometry. The mRNA expression and the protein phosphorylation levels were detected by RT-PCR and Western immunoblot assay. RESULTS: LLDT-8 at 25 and 50 nM significantly inhibited the PHA- and recall antigens-induced T cell proliferation, and suppressed mixed lymphocyte reaction. LLDT-8 reduced cytokines production (IFN-gamma, IL-2, TNF-alpha) in PHA- and Sac-activated PBMC. LLDT-8 did not alter the increased expression of MHC class I/II and B7.1, but reduced B7.2 by approximately 30%. No effect of LLDT-8 was observed for the expression of T cell activation markers (CD69, CD154). However, LLDT-8 significantly reduced IFN-gamma-expressing T cell percentages and IFN-gamma mRNA transcription in PHA-activated T cells. It also inhibited the phosphorylation levels of JNK and p38. LLDT-8 did not affect NK cytotoxic activity against K562 cells. CONCLUSION: LLDT-8 was a promising immunosuppressant for human immune-related diseases.
Subject(s)
Diterpenes/pharmacology , Immunity, Cellular/drug effects , Immunosuppressive Agents , Monocytes/drug effects , Monocytes/immunology , Antigen-Presenting Cells/drug effects , Blotting, Western , Cell Proliferation/drug effects , Cytokines/biosynthesis , Flow Cytometry , Genes, MHC Class I/drug effects , Genes, MHC Class II/drug effects , Humans , In Vitro Techniques , Interferon-gamma/biosynthesis , Killer Cells, Natural/drug effects , Killer Cells, Natural/immunology , Lymphocyte Activation/drug effects , Lymphocyte Culture Test, Mixed , Mitogen-Activated Protein Kinases/biosynthesis , Reverse Transcriptase Polymerase Chain Reaction , T-Lymphocytes/drug effects , Tetrazolium Salts , ThiazolesABSTRACT
Although lead (Pb) has significant effects on the development and function of macrophages, B cells, and T cells and has been suggested to promote allergic asthma in mice and humans, Pb modulation of bone marrow (BM)-derived dendritic cells (DCs) and the resultant DC effects on Th1 and Th2 development have not been examined. Accordingly, we cultured BM cells with murine granulocyte macrophage-colony stimulating factor (mGM-CSF)+/-PbCl(2). At day 10, culture supernatant (SN) and non-adherent cells were harvested for analysis. Additionally, day 10 non-adherent BM-DCs were harvested and recultured with mGM-CSF+LPS+/-Pb for 2 days. The day 10 Pb exposure significantly inhibited BM-DC generation, based on CD11c expression. Although fewer DCs were generated with Pb, the existing Pb-exposed DCs had significantly greater MHC-II expression than did the non-Pb-exposed DCs. However, these differences diminished upon LPS stimulation. After LPS stimulation, CD80, CD86, CD40, CD54, and MHC-II were all up-regulated on both Pb-DCs and DCs, but Pb-DCs expressed significantly less CD80 than did DCs. The CD86:CD80 ratio suggests a Pb-DC potential for Th2 cell development. After LPS stimulation, IL-6, IL-10, IL-12 (p70), and TNF-alpha levels significantly increased with both Pb-DCs and DCs, but Pb-DCs produced significantly less cytokines than did DCs, except for IL-10, which further supports Pb-DC preferential skewing toward type-2 immunity. In vitro studies confirm that Pb-DCs have the ability to polarize antigen-specific T cells to Th2 cells. Pb-DCs also enhanced allogeneic and autologous T cell proliferation in vitro, and in vivo studies suggested that Pb-DCs inhibited Th1 effects on humoral and cell-mediated immunity. The Pb effect was mainly on DCs, rather than on T cells, and Pb's modification of DC function appears to be the main cause of Pb's promotion of type-2-related immunity, which may relate to Pb's enhanced activation of the Erk/MAP kinase pathway.