| BackgroundDendritic cells (DC) as the most potent antigen presenting cells, plays a central role in the initiation and regulation of immune response.. DC can uptake, process and load antigens onto MHC molecules efficiently, It presents the antigens to T cells, By this way, DC induces tumor-specific cytotoxic T lymphocytes (CTL) and elicits protective immunity against tumor. DC-based tumor vaccines method is usually used in tumor specific immunotherapy. But human DC is not a single cell population. Besides inducing anti-tumor immunity, DC can induce tumor-special anergy or tolerance. Curative effect of the DC base-vaccines is not as good as expected, which relates to the purity of DC, the autoimmune of patients and others. Among the total, the alteration of DC subset in DC differentiation may be an important reason.DC originates from CD34~+ hematopoietic stem cells (HSC). Myeloid dendritic cells (DC1) and plasmacytoid DC (DC2) are the twoprincipal subset of human DC. Their characteristics vary a lot in phenotype,migration and function. DC1 is an effective T cells stimulator, and it can induce tumor specific immune response. In antitumor immune, the function of DC2 is uncertainty. It can stimulate tumor specific immune response. Also contribute to tumor immune tolerance. DC deriving from HSC is developing from different precursor. DC1 comes from myeloid dendritic cell precursors (pre-DC1); DC2 comes from plasmacytoid dendritic cell precursors (pre-DC2).Many researches indicate that DC in tumor microenvironment decreases in quantity and impairs in function. Otherwise, the ratio of DC1 and DC2 in tumor microenvironment changes called DC subset shift. In some researches, the phenomenon that pre-DC1/pre-DC2 in peripheral blood of patients with tumor changes and impairs in function is also observed subset shift of DC in tumor microenvironment and subset shift of pre-DC in peripheral blood of patients with tumor suggest tumor may participate in immune escape via influencing subset shift of pre-DC.Ovarian neoplasia is the first lethal cause in gynecological malignancies in women. Few symptoms in early and poor prognosis are the feature. In clinic, most of the patients with ovarian neoplasia present advanced-stage with widespread of cancer cells in the peritoneal cavity at the first diagnosis. The host's immune defect is regarded as an important cause of the metastasis of ovarian carcinoma. It's well known that the function and quantities of immune system cells in peritoneal cavity were significantly decreased and impaired, including T cells, NKs and dendritic cells.Researches of ovarian neoplasia indicate DC1 decreases in ascites; the aggregation of DC2 does not result in stimulating T cells effectively and inducing tumor specific immune response. Subset shift of pre-DC in peripheral blood of patients with ovarian neoplasia is observed. We suppose ovarian carcinoma cells may produce the subset shift of dendritic cell precursors in vitro via regulating ratio of dendretic cell precursors, which may be involved intraperitoneal immunodeficiencyIn the study, pre-DC derived from CD34~+ hematopoietic stem cells were induced by cytokine in vitro. The antigens expression of pre-DC induced by ovarian carcinoma cell lines (SKOV3) with the different concentrations was detected by flow cytometry. The aim of the study is to observe the supernate of ovarian carcinoma cells influence subset shift of dendritic cell precursors (pre-DC) in vitro.Material and methodsPre-DC derived from CD34~+hematopoietic stem cells were induced by cytokines in vitro. The concentration of cytokine FLT3-L is 100ng/ml and the concentration of cytokine SCF is 50ng/ml. We cultivate and collect the supernate of ovarian carcinoma cell lines (SKOV3) and concentrate the supernate by dialysis. The antigens expression of pre-DC was induced by the supernate of ovarian carcinoma cell lines (SKOV3) with different concentrations (0%, 25%, 50%, 75%, 100%, 150%).The antigens expression of pre-DC was detected by flow cytometry.Oneway ANOVA was used for statistical analysis. All data were managed with SPSS 13.0 for windows.Results1 The supernate of ovarian carcinoma cell lines can promote the expression of CD11c (p=0.032). When the percentage of supernate of ovarian carcinoma cell lines goes up from 0% to 50%, the expression of CD11c climbs. While the percentage of supernate of ovarian carcinoma cell lines rises from the 50% to 150%, expression level of CD11c descends compared with that of 50% (P=0.026). The supernate of ovarian carcinoma cell lines cannot promote the expression of CD123 (p=0.330). When the percentage of supernate of ovarian carcinoma cell lines goes up from 0% to 150%, expression level of CD123 increases.2 The supernate of ovarian carcinoma cell lines can promote the expression of HLA-DR (p=0.009). When the percentage of supernate of ovarian carcinoma cell lines goes up from 0% to 150%, the expression of HLA-DR increases persistently and it reaches its peak at 150% (P=0.000).3 The supernate of ovarian carcinoma cell lines cannot promote the expression of CD80 and CD86 (p=0.167 and P=0.402). When the percentage of supernate of ovarian carcinoma cell lines rises from 0% to 50%, expression level of CD80 climbs and it declines a little compared with that of 50%, when the percentage of supernate of ovarian carcinoma cell lines rises from 50% to 150%. Expressions of CD86 are mainly the same to CD80. When the percentage of supernate of ovarian carcinoma cell lines transfers from 0% to 75%, expression level of CD86 climbs and it declines a little compared with that of 75%, when the percentage of supernate of ovarian carcinoma cell lines rises from 75% to 150%.ConclusionsThe supernate of SKOV3 cell lines may produce the subset shift of dendritic cell precursors in vitro via regulating ratio of dendretic cell precursors, which may be involved intraperitoneal immunodeficiency. |
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