NBD多肽对小鼠树突状细胞成熟状态的影响

The impact of NBD peptide on the biological activity of dendritic cells cultured from murine bone marrow

目的观察NBD多肽对脂多糖(LPS)刺激的小鼠骨髓源性树突状细胞(DC)生物学活性的影响。方法培养小鼠骨髓源性DC,分为对照组、LPS组及NBD组,对照组不予任何处理,LPS组加入终质量浓度为1 mg/L的LPS,NBD组于加入NBD50μmol/ml,4 h后给予1 mg/L LPS刺激,继续培养3 d。流式细胞术检测DC细胞表面CD80、CD86及MHC-Ⅱ分子的变化,ELISA法检测各组DC分泌肿瘤坏死因子-α(TNF-α)、干扰素-γ(IFN-γ)和白细胞介素-12(IL-12)的浓度,混合淋巴细胞培养检测T细胞增殖能力,免疫细胞化学及Western blot检测DC核因子-kB(NF-kB)的表达。结果LPS可促进DC高表达CD80、CD86及MHC-Ⅱ分子,促进Th1型细胞因子释放、NF-kB高表达并易位于核内,并诱导T细胞增殖,NBD多肽可阻断LPS的这些效应。结论NBD多肽可抑制DC成熟,增强同种未成熟DC的免疫耐受诱导作用,为进一步研究DC的临床应用奠定了基础。

To observe the impact of NEMO-binding domain （NBD） peptide on the biological activity of dendritic cells （DCs） cultured from murine bone marrow and stimulated by lipopolysaccharides （LPS） ,and provide novel ideas and basis of DC＇s clinical applications. Methods Mouse DCs were generated from bone marrow cells and were divided into control group,LPS （ 1 mg/L） group and NBD （ NBD ＋ 1 mg/L LPS） group. All groups were continuously cultured for 3 days. Flow cytometry and mixed lymphocyte reaction （MLR） were used to detect the phenotype and functional properties of DCs. ELISA was used to detect the concentration of TNF-α, IFN-γ/and IL-12 in the supernatant. Immunochemistry and Western blot were used to detect the concentration of NF-KB. Results LPS stimulation increased the CD80,CD86, MHC-Ⅱ] on the cytomembrane of DCs and TNF-α, IFN-γ/, IL-12 concentration in the supernatant. LPS stimulation also increased the NF-KB and promoted the shift of NF-KB to karyon. NBD peptide could inhibit these effects. Conclusion NBD peptide can suppress the maturation of DCs and promote the toleranceinduction effect of DCs.