[Identification of dendritic cells subsets in peritoneal fluid]

Dendritic cells (DC) play a crucial role in the activation of naive T lymphocytes and in the generation of primary T cell responses. DC are present in lymphoid and non lymphoid tissues. Our previous data have shown, that DC are present in peritoneal fluid (PF). There is no other data in literature on this subject. The aim of our study was identification of DC in peritoneal fluid and peripheral blood patients with infertility (n = 5) and benign noninflammatory ovarian tumor (n = 16). Mononuclear cells from PF and peripheral blood were isolated on gradient density centrifugation (Lymphoprep, Nycomed-Norway). Isolated cells (106 of cells per tube) were incubated with mAbs. We collected 300.000 cells using a FACSCalibur flow cytometer and analysed with CellQuest Software. The following directly conjugated monoclonal antibodies were used: anti-BDCA-1(CD1c) FITC, anti-BDCA-2 FITC (Miltenyi Biotec) and anti-CD19 CyChrome, anti-CD123 PE (Pharmingen, USA). The concentrations of peritoneal fluid leukocytes in infertility patients and women with ovarian tumor approximated 6.8 x 10(6) and 7.76 x 10(6), respectively. Using this method, myeloid DC comprised 8.08 +/- 2.69% mononuclear cells PF in infertile women and 11.23 +/- 6.59% mononuclear cells PF in patients with ovarian tumor. The average mean of lymphoid DC was 0.63 +/- 0.33% mononuclear cells PF in infertile women and 0.59 +/- 0.33% mononuclear cells PF in patients with ovarian tumor. The percentage of dendritic cells in peritoneal fluid was significantly higher than in peripheral blood in both studied groups (p < 0.05).