ABSTRACT
OBJECTIVE: The aim of this study was to detect expression of different cytokines in epithelial ovarian carcinoma (EOC) cells and normal ovarian surface epithelial (OSE) cells in vitro and the levels of those with elevated expression in the EOC patients, and to analyze the contribution of cytokine profiles to tumor immune deficiency. MATERIALS AND METHODS: Cytokine antibody array was used to detect cytokine profiles in two cell lines of EOC (SKOV3 and CaoV3), primarily cultured EOC and OSE cells. The levels of leukemia inhibitory factor (LIF), interleukin-10 (IL-10), IL-4, and transforming growth factor-beta1 (TGF-beta1) in peritoneal fluids and sera in the patients with EOC and benign gynecological tumors were detected by enzyme-linked immunosorbent assay. RESULTS: The levels of LIF, IL-10, and IL-4 were detected two times higher in the culture supernatants of the EOC cell lines than those in OSE cells by cytokine antibody array. Both LIF and IL-10 levels were more increased in ascites of EOC patients than in those in benign gynecological tumor patients (P < 0.05). The level of IL-4 was not detectable in any samples of ascites or sera. No difference of TGF-beta1 value was detected between patients with EOC and benign gynecological tumors. CONCLUSION: Epithelium ovarian carcinoma cells can produce more LIF, IL-10 and IL-4 than OSE cells, and contribute to the elevated levels of those cytokines in EOC patients, which probably participates in the development of immune deficiency in the peritoneal cavity of EOC patients.
Subject(s)
Cytokines/analysis , Neoplasms, Glandular and Epithelial/immunology , Ovarian Neoplasms/immunology , Adult , Cell Line, Tumor , Female , Humans , Interleukin-10/analysis , Interleukin-4/analysis , Leukemia Inhibitory Factor/analysis , Transforming Growth Factor beta1/analysisABSTRACT
OBJECTIVE: Decreased number and impaired function of dendritic cells (DCs) have been found in ovarian carcinoma microenvironment. The study was designed to detect if this phenomenon was associated with abnormal DC differentiation influenced by ovarian carcinoma cells. MATERIALS AND METHODS: FLT-3L and SCF were used for expanding DC precursors from CD34+ progenitors. GM-CSF and TNF-alpha were used to induce mature DCs. Supernatants of cultured ovarian carcinoma cell line SKOV3 were added, in order to study their influence on the differentiation and maturation of Lin-CD45RA- DC precursors. Flow cytometry was used to analyze cell subtypes and molecular surface markers. Allogeneic T-cell proliferation assay was used to exam stimulatory activity of DCs. IL-12 secretion was tested by ELISA. RESULTS: Lin-CD45RA- DC precursors cultured with GM-CSF and TNF-alpha generated HLA-DR+CD11C+CD123- myeloid DCs (mDCs) and HLA-DR+CD11C-CD123+ plasmacytoid DCs (pDCs) in vitro. The supernatants from ovarian carcinoma cell line SKOV3 (SKOV3-supernatants) increased pDCs and decreased mDCs compared with pure medium or supernatants of normal ovarian surface epithelial (OSE) cells. There were no significantly different expressions of HLA-DR and CD80 by DCs between with and without SKOV3-supernatants. But DCs treated with SKOV3-supernatants were shown to have impaired immune activity to stimulate proliferation of allogeneic CD3+ T cells and secrete IL-12. CONCLUSION: Ovarian carcinoma cells influence differentiation of Lin-CD45RA- DC precursors into subtypes of mature DCs in vitro. This resulted in fewer mDCs, increased number of pDCs, and impairment of mature DCs immune activity.
