宫颈癌患者外周血来源的树突状细胞的生物学特性观察

Characteristics of Dendritic Cells Derived From Peripheral Blood of Patients with Invasive Cervical Carcinoma

【目的】探讨宫颈癌患者外周血来源的树突状细胞(DC)的生物学特性。【方法】14例Ib期及以上的宫颈癌患者为研究组,24例健康女性为对照组。外周血经密度梯度离心得到单个核细胞,其中附壁成分在含GM-CSF/IL-4的完全淋巴细胞培养液培养。光镜和电镜观察培养细胞的形态变化。培养7d后,计算树突状细胞的产量,流式细胞术分析HLA-ABC、HLA-DR、CD40、CD80、CD1a和CD14的表达,同种异体淋巴细胞的混合淋巴反应(MLR)检测树突状细胞刺激淋巴细胞增殖能力。【结果】经过7d培养,两组均诱导出典型形态和表型的DC。宫颈癌患者单位体积外周血的未成熟DC(ImDC)的产量为67.3×106±33.8×106,较对照组低(P<0.05)。研究组的CD40、CD80和CD1a的平均荧光强度(MFI)分别为18±9,15±6和71±11,均较对照组低(P<0.05);而两组ImDC的HLA-ABC、HLA-DR表达强度相似(P>0.05)。相同PBL/DC比值,研究组ImDC刺激同种异体淋巴细胞增殖能力下降(P<0.05)。【结论】宫颈癌患者的外周血单个核细胞培养的树突状细胞(ImDC)和健康对照组的ImDC具有免疫学差异,提示在应用树突状细胞进行免疫治疗时应考虑这种差异。

To investigate the biological characteristics of dendritic cells derived from peripheral blood of patients with advanced cervical carcinoma. 14 patients with cervical carcinoma at stages of Ib or above were assigned to study group and 24 cases of healthy females were assigned to control group. Leukocytes were isolated by density centrifugation. Adherent fractions were incubated in present of GM CSF/IL 4 for 7 days. The yield of immature DCs (ImDCs) was calculated and the morphological change of the cultured cells was observed by light microscopy and electronic microscopy. The surface expressions of HLA ABC, HLA DR, CD40, CD80, CD1a and CD14 were analysized by FCM. The ability of DCs to stimulate the proliferation of lymphocytes was determined by allogeneic MLR. Morphologically and phenologically typical DCs were obtained from two groups after culure for 7 days . The yield of ImDCs from unit volume peripheral blood (calculated as 1000ml peripheral blood)in study group was 67.3×106±33.8×106,significantly lower than that in control group(P< 0.05). The MFIs of CD40,CD80 and CD1a expressed on ImDC in study group were 18±9,15±6 and 71±11, respectively, significantly lower than that in control group(P< 0.05), while the expressions of HLA ABC, HLA DR did not differ between two groups. Moreover, the ability of DCs to stimulate the proliferation of lymphocytes in study group was lower than that in control group.[Conclusion]Our data show that ImDCs derived from monocytes of patients with cervical cancer present different immunological characteristics compared to that of healthy females. Impaired DCs' function should be considered when immunotherapy is to be performed.