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[腹膜液中树突状细胞亚群的鉴定]

[Identification of dendritic cells subsets in peritoneal fluid].

作者信息

Błaszkowska I, Roliński J, Gogacz M, Kotarski J

机构信息

I Katedry i Kliniki Ginekologii Operacyjnej Akademii Medycznej w Lublinie.

出版信息

Ginekol Pol. 2001 Dec;72(12A):1455-9.

Abstract

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).

摘要

树突状细胞(DC)在初始T淋巴细胞的激活以及原发性T细胞反应的产生中发挥着关键作用。DC存在于淋巴组织和非淋巴组织中。我们之前的数据表明,DC存在于腹腔液(PF)中。关于这个主题,文献中没有其他相关数据。我们研究的目的是鉴定不孕症患者(n = 5)和良性非炎性卵巢肿瘤患者(n = 16)的腹腔液和外周血中的DC。通过梯度密度离心法(Lymphoprep,Nycomed - 挪威)从PF和外周血中分离单核细胞。将分离的细胞(每管106个细胞)与单克隆抗体孵育。我们使用FACSCalibur流式细胞仪收集300,000个细胞,并使用CellQuest软件进行分析。使用了以下直接偶联的单克隆抗体:抗BDCA - 1(CD1c) FITC、抗BDCA - 2 FITC(Miltenyi Biotec)以及抗CD19 CyChrome、抗CD123 PE(美国Pharmingen)。不孕症患者和卵巢肿瘤女性腹腔液白细胞浓度分别约为6.8×10(6)和7.76×10(6)。使用这种方法,髓样DC在不孕女性的PF单核细胞中占8.08±2.69%,在卵巢肿瘤患者的PF单核细胞中占11.23±6.59%。淋巴样DC的平均比例在不孕女性的PF单核细胞中为0.63±0.33%,在卵巢肿瘤患者的PF单核细胞中为0.59±0.33%。在两个研究组中,腹腔液中树突状细胞的百分比均显著高于外周血中的百分比(p < 0.05)。

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