摘要
目的探讨 T 辅助(Th)1细胞免疫应答过程中,中性粒细胞(PMN)对树突状细胞(DC)的调节作用。方法将脂多糖(LPS)刺激的中性粒细胞(LPS-PMN)和未成熟树突状细胞(imDC)共培养,流式细胞术检测 DC 表面 CD40和 CD86,酶联免疫吸附试验(ELISA)检测白细胞介素12(IL-12)、肿瘤坏死囚子α(TNF-α)。将与 LPS-PMN 共培养后 DC(PMN-DC)提纯,与 FITC 标记的卵清蛋白(FITC-OVA)培养,检测其吞噬功能。将 PMN-DC 与 DO11.10T 细胞和 OVA17肽共培养,计活细胞数,胞内染色测干扰素γ(IFN-γ)、白细胞介素4(IL-4)。结果 LPS-PMN 可刺激 imDC CD40和 CD86上调并分泌 IL-12、肿瘤坏死因子(TNF)-α,且这种能力能被抗 TNF-α单抗所抑制。与 imDC埘照,PMN-DC 吞噬功能下降,能显著刺激 DO11.10T 细胞增殖,分泌高水平 IFN-γ、少量 IL-4。结论LPS-PMN 具有促使 imDC 成熟活化,刺激其分泌细胞因子,发挥其在 Th1免疫应答中抗原呈递,促进 Th1分化的作用。
Objective To investigate whether polymorphonuclear nentrophilic leucocyte （PMN） can induce the maturation and activation of immature dendritic cells, thus promoting the Thl immunity to microbial pathogens in vitro. Methods PMN isolated from mouse bone marrow were stimulated by lipopolysaccharide （LPS） , and were cultured with immature DC （imDCs） isolated from mouse bone marrow for 18 hours. Then the analysis of DC surface markers CD40 and CD86 was done by flow cytometry, and ELISA was used to detect the levels of TNF-α and IL-12. Purified DC cocultured with LPS-PMN （PMN-DC） and imDC were incubated with fluorescein isothiocyanate conjugated OVA （FITC-OVA） , and the phagocytic capacity of PMN-DC and imDC was assessed by flow cytometry. Splenic CD4^＋ T cells from DO11.10×C57BL/6 F1 hybrid mice were obtained, and then cultured with PMN-DC or imDC in the presence of OVA （323-339）. The cells were double stained with anti-CD4-PE and 7-amino-actinomycin D. The cellular data were acquired for 56s with a flow cytometer and the assay of IFN-γ and IL-4 was done by intracellular staining. Results LPS-PMN induced strong up-regulation of CD40 and CD86, and this capacity was remarkably inhibited after being neutralized by anti-TNF-α monoclonal antibody. LPS-PMN also stimulated imDCs to produce IL-12 and TNF-α. PMN-DCs demonstrated decreased phagocytic capacity compared with that of imDCs. Furthermore, the PMN-DC induced considerable DO11. 10 T cell proliferation, and stimulated DO11. 10 T cell to produce a large amount of IFN-γ but a relatively low amount of IL-4. Conclusion LPS-PMN can induce the maturation and activation of imDCs, including the cytokine secretion, specific antigen presentation and Thl differentiation, thus promoting the type Ⅰ immunity to microbial pathogens.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2006年第39期2761-2765,共5页
National Medical Journal of China
基金
上海市科学技术委员会资助项目(054119639)
关键词
粒细胞
树突细胞
免疫
细胞
Granulocytes
Dendritic cells
Immunity, cellular
作者简介
通讯作者：陈刚，Email：chgan305@163．com
