ABSTRACT
BACKGROUND: Ayurved Siriraj Wattana recipe (AVS073), has been prescribed as tonic, to increase appetite, and for pain relief. It also exhibits antioxidant, anti-inflammatory, immunomodulating and anti-cancer activities. However, the immunomodulatory effects on antigen-presenting cells and effector T cells remained elusive. We thus aimed to study the effects of AVS073 on differentiation, maturation, functions and proportions of CIK cells and monocyte-derived DCs. METHODS: CIK cells and monocyte-derived DCs were treated with AVS073, followed by the assessment of T-helper (Th) phenotypes using real-time RT-PCR and flow cytometry. RESULTS: AVS073 promoted Th1 phenotype in CD3+CD56+ subset of CIK cells through increasing STAT4, T-bet, and interferon-γ. AVS073 inhibited Th2 phenotype through decreasing STAT6. AVS073 inhibited Treg phenotype through decreasing STAT5A, STAT5B and IDO. AVS073 promoted Th17 phenotype through increasing STAT3, RORC and IL-17. AVS073 treatment of mDCs resulted in increasing Th1-prone cytokine (IL-12) and Th17-prone cytokines (IL-6 and IL-23). CONCLUSIONS: AVS073 upregulated Th1 and Th17, but downregulated Th2 and Treg phenotypes within CD3+CD56+ cells. The treatment of mDCs drove Th1 and Th17-polarizations.
Subject(s)
Cytokine-Induced Killer Cells/drug effects , Dendritic Cells/drug effects , Plant Preparations/pharmacology , Plants, Medicinal/chemistry , CD3 Complex , CD56 Antigen , Cell Proliferation/drug effects , Cell Survival/drug effects , Cells, Cultured , Cytokines/metabolism , Humans , Immunophenotyping , T-Lymphocyte Subsets/drug effects , ThailandABSTRACT
Cytokine-induced killer (CIK) cells have reached clinical trials for leukemia and solid tumors. Their anti-tumor cytotoxicity had earlier been shown to be intensified after the co-culture with dendritic cells (DCs). We observed markedly enhanced anti-tumor cytotoxicity activity of CIK cells after the co-culture with sunitinib-pretreated DCs over that of untreated DCs. This cytotoxicity was reliant upon DC modulation by sunitinib because the direct exposure of CIK cells to sunitinib had no significant effect. Sunitinib promoted Th1-inducing and pro-inflammatory phenotypes (IL-12, IFN-γ and IL-6) in DCs at the expense of Th2 inducing phenotype (IL-13) and regulatory phenotype (PD-L1, IDO). Sunitinib-treated DCs subsequently induced the upregulation of Th1 phenotypic markers (IFN-γ and T-bet) and the downregulation of the Th2 signature (GATA-3) and the Th17 marker (RORC) on the CD3âºCD56⺠subset of CIK cells. It concluded that sunitinib-pretreated DCs drove the CD3âºCD56⺠subset toward Th1 phenotype with increased anti-tumor cytotoxicity.
Subject(s)
Antineoplastic Agents/pharmacology , CD3 Complex/metabolism , CD56 Antigen/metabolism , Cytokine-Induced Killer Cells/immunology , Dendritic Cells/immunology , Indoles/pharmacology , Pyrroles/pharmacology , Cell Line, Tumor , Coculture Techniques , Cytokine-Induced Killer Cells/drug effects , Cytokine-Induced Killer Cells/metabolism , Dendritic Cells/drug effects , Dendritic Cells/metabolism , Drug Screening Assays, Antitumor , Flow Cytometry , Humans , Interferon-gamma/antagonists & inhibitors , Interferon-gamma/metabolism , Macrophages/drug effects , Macrophages/metabolism , Sunitinib , T-Lymphocytes, Regulatory/drug effects , T-Lymphocytes, Regulatory/immunologyABSTRACT
Cytokine-induced killer (CIK) cells were examined for safety and efficacy for cholangiocarcinoma treatment. Several conditions of human CIK cells were examined using ex vivo cytotoxic assay and SCID mice pre-inoculated with cholangiocarcinoma cells. We monitored the ex vivo cytotoxicity, tumor sizes and immunohistochemistry. Optimal tumor suppression was observed when CIK cells were pre-exposed to dendritic cells (DCs). Unexpectedly, pulsing of tumor RNA to DCs rendered the co-culturing CIK cells ineffective and raised the proportion of CD4(+)CD25(+) subset. The use of CD3(+)CD56(+) subset instead of the whole population of CIK cells for the co-culture with RNA-pulsed DCs restored the efficacy. Tumor-infiltrating human CD3(+) cells were observed from day 2 - 14. The CD3(+)CD56(+) cells are logical candidates for clinical trial while the DC-co-cultured CIK cells produced similar efficacy and more feasible for clinical application. The RNA pulsation of DCs up-regulated the regulatory subset of CIK cells and abrogated the anti-tumor efficacy.
Subject(s)
Bile Duct Neoplasms/therapy , Cholangiocarcinoma/therapy , Cytokine-Induced Killer Cells/immunology , Animals , Bile Duct Neoplasms/pathology , Bile Ducts, Intrahepatic , CD3 Complex/analysis , CD56 Antigen/analysis , Cell Line, Tumor , Cholangiocarcinoma/pathology , Dendritic Cells/immunology , Humans , Mice , Mice, SCID , Neoplasm Transplantation , Transplantation, HeterologousABSTRACT
BACKGROUND: Indoleamine 2, 3-dioxygenase (IDO), a tryptophan-degrading enzyme in dendritic cells (DCs), mediates an immunosuppressive effect on activated T lymphocytes. However, little is known about the effect of Der p 1 on IDO in human DCs. OBJECTIVE: The aim was to investigate the effect of Der p 1 on the expression and activity of IDO in monocyte-derived DCs from house dust mite (HDM)-sensitive patients with asthma. METHODS: Using real-time RT-PCR and HPLC, the expression and activity of IDO were assessed in TNF-alpha-induced mature DCs from HDM-sensitive and nonatopic patients with asthma in response to Der p 1 exposure ex vivo. We also monitored the alteration of IDO activity in Der p 1-pulsed DCs after the coincubation with autologous T cells. RESULTS: With a reliance on its protease activity, Der p 1 suppressed functional IDO in DCs from HDM-sensitive patients with asthma but enhanced IDO activity in DCs from nonatopic patients with asthma. This suppression was maintained by the reciprocally induced IL-4 from the coculturing autologous HDM-sensitive T cells. Conversely, the upregulation of IDO activity in Der p 1-pulsed DCs was maintained by IFN-gamma released from autologous nonatopic T cells and the regulatory T-cell subset. Der p 1 pulsation to sensitive DCs failed to raise regulatory T cells but raised progenitor fractions from cloned HDM-sensitive CD4(+) cells through direct contact and soluble mediators. CONCLUSION: House dust mite-sensitive DCs exposed to Der p 1 downregulated IDO activity and tipped the T(H)1/T(H)2 cytokine balance toward IL-4, resulting in sustainable IDO suppression.
Subject(s)
Antigens, Dermatophagoides/immunology , Asthma/immunology , Dendritic Cells/immunology , Gene Expression Regulation, Enzymologic/immunology , Indoleamine-Pyrrole 2,3,-Dioxygenase/immunology , T-Lymphocytes, Helper-Inducer/immunology , Antigens, Dermatophagoides/pharmacology , Arthropod Proteins , Asthma/enzymology , Cells, Cultured , Cysteine Endopeptidases , Dendritic Cells/enzymology , Female , Gene Expression Regulation, Enzymologic/drug effects , Humans , Indoleamine-Pyrrole 2,3,-Dioxygenase/biosynthesis , Interferon-gamma/immunology , Interleukin-4/immunology , Male , T-Lymphocytes, Helper-Inducer/enzymology , Tumor Necrosis Factor-alpha/immunology , Up-Regulation/drug effects , Up-Regulation/immunologyABSTRACT
OBJECTIVE: To determine if telomerase activity can differentiate malignant from tuberculous pleural effusions. DESIGN: Telomerase activity in malignant and tuberculous pleural effusions was measured in a blinded manner using a PCR-based telomeric repeat amplification protocol (TRAP) assay. MATERIAL AND METHOD: Fifty-two patients with lymphocytic exudative pleural effusions were identified on thoracocentasis over a period of 18 months. RESULTS: Telomerase activity was detected in 34% of malignant pleural fluid samples and 50% of tuberculous pleural effusions. The positive rate of telomerase activity was 30.7% for primary lung cancer and 37.5% for metastatic pleural effusion. The sensitivity and specificity of telomerase activity assay were extremely low (35.7% and 52.9%, respectively), compared with that of cytological examination (52.6% and 65.4%, respectively). Moreover the diagnostic accuracy of telomerase activity in combination with cytology was even lower than cytological examination alone (46.7% vs. 60%, respectively). This finding was in contrast to previous reports and demonstrated that the detection rate of telomerase activity in tuberculous pleural effusions was greater than that observed in malignant pleural exudates. CONCLUSION: Telomerase activity does not appear to be a useful marker for differentiating malignant from tuberculous effusions.