title: Attenuated in vitro effects of IFN-α, IL-2 and IL-12 on functional and receptor characteristics of peripheral blood lymphocytes in metastatic melanoma patients.
abstract: Considering tumor-induced suppression of lymphocytes the aim of this study was to investigate in vitro effects of IFN-α, IL-2, IL-12 and IL-18 as immunomodulating agents on the functional and receptor characteristics of peripheral blood lymphocytes (PBL) in metastatic melanoma (MM) patients compared to healthy controls (HC). In HC IFN-α, IL-2 and IL-12 enhanced mRNA level of perforin by inducing pSTAT-1 and pSTAT-5 signaling molecules. Additionally, the expression of NKG2D activating receptor and its DAP10 signaling molecule was upregulated by IL-2. Contrary to this, in MM patients only IL-2 by upregulating pSTAT-5 increased perforin-mediated cytotoxicity of lymphocytes. Furthermore, there was significantly negative correlation between the percentage of CD4+CD25bright+CD27+regulatory T (Treg) cells and NK cell cytotoxicity, as well as the expression of NKG2D receptor on PBL in HC and MM patients. Therefore, the absence of IL-2 effect on the increase of NKG2D/DAP10 level in MM patients could be the consequence of the increased percentage of immunosuppressive CD4+CD25bright+CD27+cells after this cytokine treatment in patients. However, in MM IL-12 significantly decreases the percentage of these inhibitory cells. Although IL-2 as a single agent has numerous side effects, it remains the important cytokine for PBL activation in melanoma immunotherapy. Additionally, the removal of Treg cells from patient PBL by IL-12 before in vitro stimulation with IL-2, may lead to the generation of more potent cytotoxic lymphocytes against tumor cells. Therefore, lymphocyte based therapy for MM patients should integrate not only the choice of appropriate immunostimulatory cytokine, but also the removal of inhibitory cells from tumor microenvironment.
authors.affiliation: Department of Experimental Oncology, Institute of Oncology and Radiology of Serbia, Pasterova 14, 11000 Belgrade, Serbia.
authors.forename: Katarina M
authors.initials: KM
authors.lastname: Mirjačić Martinović
authors.affiliation: Department of Experimental Oncology, Institute of Oncology and Radiology of Serbia, Pasterova 14, 11000 Belgrade, Serbia.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Vuletić
authors.affiliation: Department of Medical Oncology, Institute of Oncology and Radiology of Serbia, Pasterova 14, 11000 Belgrade, Serbia.
authors.forename: Nada
authors.initials: N
authors.lastname: Lj Babović
authors.affiliation: Surgical Oncology Clinic, Institute of Oncology and Radiology of Serbia, Pasterova 14, 11000 Belgrade, Serbia; School of Medicine, University of Belgrade, Dr Subotića 8, 11000 Beograd, Serbia.
authors.forename: Radan R
authors.initials: RR
authors.lastname: Džodić
authors.affiliation: Department of Experimental Oncology, Institute of Oncology and Radiology of Serbia, Pasterova 14, 11000 Belgrade, Serbia; School of Medicine, University of Belgrade, Dr Subotića 8, 11000 Beograd, Serbia.
authors.forename: Gordana M
authors.initials: GM
authors.lastname: Konjević
authors.affiliation: Faculty of Medical Sciences, University of Kragujevac, P. Box 124, 34000 Kragujevac, Serbia. Electronic address: vdvd@lycos.com.
authors.forename: Vladimir B
authors.initials: VB
authors.lastname: Jurišić
mesh_terms: D000328:Adult; D000368:Aged; D003601:Cytotoxicity Tests, Immunologic; D005260:Female; D006801:Humans; D056747:Immunomodulation; D016898:Interferon-alpha / Q000494:pharmacology*; D018664:Interleukin-12 / Q000494:pharmacology*; D020382:Interleukin-18 / Q000494:pharmacology; D007376:Interleukin-2 / Q000494:pharmacology*; D007963:Leukocytes, Mononuclear / Q000187:drug effects*; D008213:Lymphocyte Activation; D008297:Male; D008545:Melanoma / Q000097:blood*; D008875:Middle Aged; D055655:NK Cell Lectin-Like Receptor Subfamily K; D009362:Neoplasm Metastasis; D054353:Perforin / Q000235:genetics / Q000378:metabolism; D050794:STAT1 Transcription Factor / Q000235:genetics; D050799:STAT5 Transcription Factor / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects; D050378:T-Lymphocytes, Regulatory / Q000187:drug effects
chemical_list: C000589760:IL18 protein, human; D016898:Interferon-alpha; D020382:Interleukin-18; D007376:Interleukin-2; D055655:NK Cell Lectin-Like Receptor Subfamily K; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D050799:STAT5 Transcription Factor; D054353:Perforin; D018664:Interleukin-12
keywords: Cytotoxicity; IFN-α/IL-2/IL-12/IL-18; Lymphocytes; Metastatic melanoma