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  • Title: Ovarian carcinoma cells influence differentiation of Lin-CD45RA- dendritic cell precursors into two mature subtypes in vitro.
    Author: Chen LL, Ye F, Yu Y, Chen HZ, Zhang WW, Lü WG, Xie X.
    Journal: Gynecol Oncol; 2009 Jan; 112(1):199-204. PubMed ID: 19022493.
    Abstract:
    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.
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