BACKGROUND: Dendritic cell (DC)-based vaccines are currently being evaluated as a novel strategy for tumor vaccination and immunotherapy. However, inducing long-term regression in established tumor-implanted mice is difficult. Here, we show that deoxypohophyllotoxin (DPT) induces maturation and activation of bone marrow-derived DCs via Toll-like receptor (TLR) 4 activation of MAPK and NF-kappaB. METHODS: The phenotypic and functional maturation of DPT-treated DCs was assessed by flow cytometric analysis and cytokine production, respectively. DPT-treated DCs was also used for mixed leukocyte reaction to evaluate T cell-priming capacity and for tumor regression against melanoma. RESULTS: DPT promoted the activation of CD8+ T cells and the Th1 immune response by inducing IL-12 production in DCs. In a B16F10 melanoma-implanted mouse model, we demonstrated that DPT-treated DCs (DPT-DCs) enhance immune priming and regression of an established tumor in vivo. Furthermore, migration of DPT-DCs to the draining lymph nodes was induced via CCR7 upregulation. Mice that received DPT-DCs displayed enhanced antitumor therapeutic efficacy, which was associated with increased IFN-gamma production and induction of cytotoxic T lymphocyte activity. CONCLUSION: These findings strongly suggest that the adjuvant effect of DPT in DC vaccination is associated with the polarization of T effector cells toward a Th1 phenotype and provides a potential therapeutic antitumor immunity.
Animals ; Dendritic Cells ; Immunotherapy ; Interleukin-12 ; Lymph Nodes ; Lymphocyte Culture Test, Mixed ; Lymphocytes ; Mice ; Phenotype ; Podophyllotoxin ; T-Lymphocytes ; Toll-Like Receptors ; Up-Regulation ; Vaccination ; Vaccines

BACKGROUND: Graft-versus-host disease (GVHD) is a huddle for success of hematopoietic stem cell transplantation. In this study, effects of irradiation dose on immune kinetics of GVHD were investigated using B6 --> BALB.B system, a mouse model for GVHD after MHC-matched allogeneic transplantation. METHODS: BALB.B mice were transplanted with bone marrow and spleen cells from C57BL/6 mice after irradiation with different doses. Leukocytes residing in the peripheral blood and target organs were collected periodically from the GVHD hosts for analysis of chimerism formation and immune kinetics along the GVHD development via flow cytometry. Myeloid cells were tested for production of IL-17 via flow cytometry. RESULTS: Pre-conditioning of BALB.B hosts with 900 cGy and 400 cGy resulted in different chimerism of leukocytes from the blood and affected survival of GVHD hosts. Profiles of leukocytes infiltrating GVHD target organs, rather than profiles of peripheral blood leukocytes (PBLs), were significantly influenced by irradiation dose. Proportions of IL-17 producing cells in the infiltrating Gr-1(+) or Mac-1(+) cells were higher in the GVHD hosts with high does irradiation than those with low dose irradiation. CONCLUSION: Pre-conditioning dose affected tissue infiltration of leukocytes and cytokine production by myeloid cells in the target organs.
Animals ; Bone Marrow ; Chimerism ; Flow Cytometry ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Interleukin-17 ; Kinetics ; Leukocytes ; Mice ; Myeloid Cells ; Spleen ; Transplants

BACKGROUND: Insulin resistance is an integral feature of metabolic syndromes, including obesity, hyperglycemia, and hyperlipidemia. In this study, we evaluated whether the aloe component could reduce obesity-induced inflammation and the occurrence of metabolic disorders such as blood glucose and insulin resistance. METHODS: Male C57BL/6 obese mice fed a high-fat diet for 54 days received a supplement of aloe formula (PAG, ALS, Aloe QDM, and Aloe QDM complex) or pioglitazone (PGZ) and were compared with unsupplemented controls (high-fat diet; HFD) or mice fed a regular diet (RD). RT-PCR and western blot analysis were used to quantify the expression of obesity-induced inflammation. RESULTS: Aloe QDM lowered fasting blood glucose and plasma insulin compared with HFD. Obesity-induced inflammatory cytokine (IL-1beta, -6, -12, TNF-alpha) and chemokine (CX3CL1, CCL5) mRNA and protein were decreased markedly, as was macrophage infiltration and hepatic triglycerides by Aloe QDM. At the same time, Aloe QDM decreased the mRNA and protein of PPARgamma/LXRalpha and 11beta-HSD1 both in the liver and WAT. CONCLUSION: Dietary aloe formula reduces obesity-induced glucose tolerance not only by suppressing inflammatory responses but also by inducing anti-inflammatory cytokines in the WAT and liver, both of which are important peripheral tissues affecting insulin resistance. The effect of Aloe QDM complex in the WAT and liver are related to its dual action on PPARgamma and 11beta-HSD1 expression and its use as a nutritional intervention against T2D and obesity-related inflammation is suggested.
11-beta-Hydroxysteroid Dehydrogenase Type 1 ; Aloe ; Animals ; Blood Glucose ; Blotting, Western ; Cytokines ; Diabetes Mellitus, Type 2 ; Diet ; Diet, High-Fat ; Fasting ; Glucose ; Humans ; Hyperglycemia ; Hyperlipidemias ; Inflammation ; Insulin ; Insulin Resistance ; Liver ; Macrophages ; Male ; Mice ; Mice, Obese ; Obesity ; Plasma ; PPAR gamma ; RNA, Messenger ; Thiazolidinediones ; Triglycerides

BACKGROUND: Epstein-Barr virus associated gastric lymphoepithelioma-like carcinoma (LELC) is characterized by the intensive infiltration of lymphoid cells, the presence of EBV, and the better prognosis over typical adenocarcinoma. Thus, it was assumable that viral latent proteins may be responsible for the recruitment of a certain T cell repertoire to EBV-associated gastric carcinoma. METHODS: To examine above possibility, EBV gene expression in gastric carcinoma tissues and usage of TCR among the tumor infiltrating lymphocytes were analyzed. RESULTS: EBV specific DNA and EBERs RNA were detected in 4 out of 30 patients. RT-PCR analysis revealed that all 4 of EBV-positive tumor tissues expressed EBNA1 mRNA and BARTs and LMP2a was detected only one sample out of 4. However, the EBNA2 and LMP-1 transcripts were not detected in these tissues. CD8+ T cells were the predominant population of infiltrating lymphocytes in the EBV-positive gastric carcinoma. According to spectra type analysis of infiltrating T cells, 10 predominant bands were detected by TCR Vbeta CDR3 specific RT-PCR from 4 EBV-positive tumor tissues. Sequence analysis of these bands revealed oligoclonal expansion of T cells. CONCLUSION: These findings suggest that clonally expanded T cells in vivo might be a population of cytotoxic T cells reactive to EBV-associated gastric carcinoma.
Adenocarcinoma ; DNA ; Gene Expression ; Herpesvirus 4, Human ; Humans ; Lymphocytes ; Lymphocytes, Tumor-Infiltrating ; Prognosis ; Proteins ; RNA ; RNA, Messenger ; Sequence Analysis ; T-Lymphocytes

BACKGROUND: In this study, we have investigated the effect of Korean red ginseng (KRG) extracts on the production of TNF-alpha and IL-8 in human keratinocytes. Also, to examine the antioxidative effect of red ginseng extracts, free radical scavenging activity and superoxide dismutase (SOD) activity in human dermal fibroblasts was measured. METHODS: To investigate the effect of KRG in atopic dermatitis, we measured the level of TNF-alpha and IL-8 secretion in LPS-stimulated human keratinocytes after the treatment of KRG extracts using enzyme-linked immunosorbent assay. Anti-oxidative activity was investigated by measuring 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and SOD activity. RESULTS: The stimulation of human keratinocytes with KRG extracts shifted the LPS-induced cytokine secretion toward a more immunosuppressive response. KRG dose-dependently decreased TNF-alpha and IL-8 production in HaCaT cells and a significant inhibition of TNF-alpha was shown when cells were treated with 500 and 1,000 microg/ml of KRG extracts. Additionally, KRG extracts showed DPPH radical scavenging and SOD activity in a dose-dependent manner. Particularly, SOD activities of concentrations higher than 60 microg/ml of KRG extracts were significantly different in human dermal fibroblast cells. CONCLUSION: Based on this study, KRG extracts may be a useful immunosuppressive agent in the treatment of atopic dermatitis.
Biphenyl Compounds ; Dermatitis, Atopic ; Enzyme-Linked Immunosorbent Assay ; Fibroblasts ; Humans ; Interleukin-8 ; Keratinocytes ; Panax ; Picrates ; Superoxide Dismutase ; Tumor Necrosis Factor-alpha

The discovery of microRNA (miRNA) is one of the major scientific breakthroughs in recent years and has revolutionized current cell biology and medical science. miRNAs are small (19~25nt) noncoding RNA molecules that post-transcriptionally regulate gene expression by targeting the 3' untranslated region (3'UTR) of specific messenger RNAs (mRNAs) for degradation of translation repression. Genetic ablation of the miRNA machinery, as well as loss or degradation of certain individual miRNAs, severely compromises immune development and response, and can lead to immune disorders. Several sophisticated regulatory mechanisms are used to maintain immune homeostasis. Regulatory T (Treg) cells are essential for maintaining peripheral tolerance, preventing autoimmune diseases and limiting chronic inflammatory diseases. Recent publications have provided compelling evidence that miRNAs are highly expressed in Treg cells, that the expression of Foxp3 is controlled by miRNAs and that a range of miRNAs are involved in the regulation of immunity. A large number of studies have reported links between alterations of miRNA homeostasis and pathological conditions such as cancer, cardiovascular disease and diabetes, as well as psychiatric and neurological diseases. Although it is still unclear how miRNA controls Treg cell development and function, recent studies certainly indicate that this topic will be the subject of further research. The specific circulating miRNA species may also be useful for the diagnosis, classification, prognosis of diseases and prediction of the therapeutic response. An explosive literature has focussed on the role of miRNA. In this review, I briefly summarize the current studies about the role of miRNAs in Treg cells and in the regulation of the innate and adaptive immune response. I also review the explosive current studies about clinical application of miRNA.
3' Untranslated Regions ; Adaptive Immunity ; Autoimmune Diseases ; Cardiovascular Diseases ; Gene Expression ; Homeostasis ; Immune System Diseases ; MicroRNAs ; Peripheral Tolerance ; Prognosis ; Repression, Psychology ; RNA, Messenger ; RNA, Untranslated ; T-Lymphocytes, Regulatory

Interleukin-7 (IL-7) is an essential cytokine for T cells. However, IL-7 is not produced by T cells themselves such that T cells are dependent on extrinsic IL-7. In fact, in the absence of IL-7, T cell development in the thymus as well as survival of naive T cells in the periphery is severely impaired. Furthermore, modulating IL-7 availability in vivo either by genetic means or other experimental approaches determines the size, composition and function of the T cell pool. Consequently, understanding IL-7 expression is critical for understanding T cell immunity. Until most recently, however, the spatiotemporal expression of in vivo IL-7 has remained obscured. Shortage of such information was partly due to scarce expression of IL-7 itself but mainly due to the lack of adequate reagents to monitor IL-7 expression in vivo. This situation dramatically changed with a recent rush of four independent studies that describe the generation and characterization of IL-7 reporter mice, all utilizing bacterial artificial chromosome transgene technology. The emerging consensus of these studies confirmed thymic stromal cells as the major producers of IL-7 but also identified IL-7 reporter activities in various peripheral tissues including skin, intestine and lymph nodes. Strikingly, developmental and environmental cues actively modulated IL-7 reporter activities in vivo suggesting that IL-7 regulation might be a new mechanism of shaping T cell development and homeostasis. Collectively, the availability of these new tools opens up new venues to assess unanswered questions in IL-7 biology in T cells and beyond.
Animals ; Biology ; Chromosomes, Artificial, Bacterial ; Consensus ; Cues ; Homeostasis ; Indicators and Reagents ; Interleukin-7 ; Intestines ; Lymph Nodes ; Mice ; Organothiophosphorus Compounds ; Skin ; Stromal Cells ; T-Lymphocytes ; Thymus Gland ; Transgenes

BACKGROUND: CM1 (centrocyte/-blast marker 1) is originally defined as a germinal center B cell marker. It is known that CM1 plays a critical role on B cell development in germinal center. In addition, we have found that CM1 is expressed on lymphoma cell lines, such as Raji, Ramos and IM-9. This means that CM1 might be served as a tumor marker as well. In the present study, we examined the expression of CM1 on the surface of the other tumors and the possibility of the development of tumor screening ELISA kit by using CM1. METHODS: First, we have examined the expression of CM1 on stomach cancer and hepatoma, which are predominantly (discovered) occurred in Korean, by flow cytometry analysis. After purifying of CM1 antigen from Raji and Ramos, the optimal ELISA condition was determined. And then we compared the level of CM1 between normal individuals and cancer patients by ELISA. To decrease the non-specific binding of anti-CM1 mAb with serum components except CM1 and to enhance the diagnostic accuracy, albumin depletion spin column was used. RESULTS: CM1 was highly expressed on stomach cancer and hepatoma cell lines. In addition, we have also confirmed the increased CM1 expression on cancer patients. The difference of CM1 expression between normal individuals and cancer patients were more clearly observed, after deletion of serum albumin by using albumin depletion spin column. CONCLUSION: Based on the results from this study, CM1 might be a useful molecule for the early diagnosis of cancer. In addition, further studies for the increase of ELISA sensitivity and appropriate albumin depletion methods should be needed.
Carcinoma, Hepatocellular ; Cell Line ; Early Detection of Cancer ; Enzyme-Linked Immunosorbent Assay* ; Flow Cytometry ; Germinal Center ; Humans ; Lymphoma ; Mass Screening* ; Serum Albumin ; Stomach Neoplasms

The immune response to any stimulus is complex, requiring coordinated action by several types of cells in a tightly regulated sequence. Thus, a physical stress such as exercise may act at any number of points in the complex sequence of events collectively termed the immune response. Although exercise causes many propound changes in parameters of immune function, the nature and magnitude of such changes rely on several factors including the immune parameters of interest; type, intensity, and duration of exercise; fitness level or exercise history of the subject; environmental factors such as ambient temperature and humidity. Although regular moderate exercise appears to be important factor for increasing immunity, Athletes are susceptible to illness, in particular upper respiratory track infection, during periods of intense training and after competition. In addition, in elite athletes, frequent illness is associated with overtraining syndrome, a neuroendocrine disorder resulting from excessive training. Through this paper, we want to investigate the effects of exercise on the immunosuppression such as exercise induced lymphopenia, asthma, anaphylaxis, URT (upper respiratory track), and TB (tuberculosis) infection. and also, we want to suggest a direct mechanism, protection and therapy of exercise induced immunosuppression.
Anaphylaxis ; Asthma ; Athletes ; Humans ; Humidity ; Immunosuppression ; Lymphopenia

BACKGROUND: As an attempt to develop a strategy to improve the protective immune response to GM-CSF-secreting CT26 (GM-CSF/CT26) tumor vaccine, we have investigated whether the apoptogenic treatment of GM-CSF/CT26 prior to vaccination enhances the induction of anti-tumor immune response in mouse model. METHODS: A carcinogen- induced mouse colorectal tumor, CT26 was transfected with GM-CSF gene using a retroviral vector to generate GM-CSF-secreting CT26 (CT26/GM-CSF). The CT26/GM-CSF was treated with gamma-irradiation or mitomycin C to induce apoptosis and vaccinated into BALB/c mice. After 7 days, the mice were injected with a lethal dose of challenge live CT26 cells to examine the protective effect of tumor vaccination in vivo. RESULTS: Although both apoptotic and necrotic CT26/GM-CSF vaccines were able to enhance anti-tumor immune response, apoptotic CT26/GM-CSF induced by pretreatment with gamma-irradiation (50,000 rads) was the most potent in generating the anti-tumor immunity, and thus 100% of mice vaccinated with the apoptotic cells remained tumor free for more than 60 days after tumor challenge. CONCLUSION: Apoptogenic pretreatment of GM-CSF-secreting CT26 tumor vaccine by gamma-irradiation (50,000 rads) resulted in a significant enhancement in inducing the protective anti-tumor immunity. A rapid induction of apoptosis of CT26/GM-CSF tumor vaccine at the vaccine site might be critical for the enhancement in anti-tumor immune response to tumor vaccine.
Animals ; Apoptosis ; Colony-Stimulating Factors* ; Colorectal Neoplasms ; Granulocyte-Macrophage Colony-Stimulating Factor ; Mice ; Mitomycin ; Vaccination ; Vaccines ; Zidovudine