title: IL-10 production in non-small cell lung carcinoma patients is regulated by ERK, P38 and COX-2.
abstract: Immune dysfunction is hallmark of patients with non-small cell lung carcinoma (NSCLC). The molecular mechanism involved in COX-2- and PGE2-mediated production of immunosuppressive cytokine IL-10 is not well-understood. Our study addresses the involvement of T cell downstream signalling intermediates, cytokines (IL-10 and IFN-γ) and their transcription factors (T-bet and GATA-3) in COX-2-mediated regulation of lymphocyte functions in NSCLC patients. In comparison to healthy individual, a marked decrease in lymphocyte proliferation to anti-CD3 MAb was observed in NSCLC patients by thymidine incorporation assay. Using flow cytometry, decrease in intracellular calcium release with increase in reactive oxygen species was observed in lymphocytes of NSCLC patients. These patients showed increased IL-10 and PGE2 with reduced IFN-γ production by ELISA. Results demonstrated defect in regulation of transcription factors T-bet and GATA-3 as analysed by Western blotting (WB), immunoprecipitation and EMSA. Overexpression of p-p38, p-ERK and COX-2 were observed with diminished p-JNK by WB. IL-10/IFN-γ levels were found to be differentially regulated via p38 and ERK mitogen-activated protein kinase (MAPK) pathways in cooperation with COX-2. Inhibition of these pathways using selective inhibitors lead to increased lymphocyte proliferative response to anti-CD3 MAb and IFN-γ production with decrease in IL-10 production. Studies showed involvement of ERK, p38 and COX-2 pathways in high IL-10 production, driven by lung tumour derived PGE2. The selective COX-2 inhibitor rofecoxib showed ability to alter the cytokine balance by affecting regulation of T-bet and GATA-3 transcription factors.
authors.affiliation: Chiplunkar Lab, Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre, Navi Mumbai, India.
authors.forename: Swati
authors.initials: S
authors.lastname: Patel
authors.forename: Shamal
authors.initials: S
authors.lastname: Vetale
authors.forename: Pradeep
authors.initials: P
authors.lastname: Teli
authors.forename: Rajesh
authors.initials: R
authors.lastname: Mistry
authors.forename: Shubhada
authors.initials: S
authors.lastname: Chiplunkar
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy / Q000235:genetics / Q000276:immunology*; D049109:Cell Proliferation / Q000187:drug effects; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000502:physiology*; D052246:Cyclooxygenase 2 Inhibitors / Q000494:pharmacology; D015232:Dinoprostone / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000502:physiology*; D050990:GATA3 Transcription Factor / Q000235:genetics / Q000276:immunology; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis / Q000276:immunology; D016753:Interleukin-10 / Q000096:biosynthesis* / Q000276:immunology; D007783:Lactones / Q000494:pharmacology*; D008175:Lung Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000276:immunology*; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology; D061251:Primary Cell Culture; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013450:Sulfones / Q000494:pharmacology*; D020825:T-Box Domain Proteins / Q000235:genetics / Q000276:immunology; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics / Q000502:physiology*; D000098815:T-bet Transcription Factor
chemical_list: D000911:Antibodies, Monoclonal; D052246:Cyclooxygenase 2 Inhibitors; D050990:GATA3 Transcription Factor; C494749:GATA3 protein, human; D007783:Lactones; D017382:Reactive Oxygen Species; D013450:Sulfones; D020825:T-Box Domain Proteins; D000098815:T-bet Transcription Factor; C116926:rofecoxib; D016753:Interleukin-10; D007371:Interferon-gamma; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D048049:Extracellular Signal-Regulated MAP Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D015232:Dinoprostone