Mohamadzadeh M, Knop J, Aliani S, Cruz P D
Department of Dermatology, University of Mainz, Germany.
Arch Dermatol Res. 1997 Jul;289(8):435-9. doi: 10.1007/s004030050217.
We took advantage of the recently generated 4F7 mAb, which recognizes an epitope expressed on dendritic cells (DC) from different tissues, to freshly isolate and positively sort for these cells and to characterize their cytokine pattern and antigen-presenting capacity in comparison with epidermal Langerhans cells (LC). RT-PCR and Northern blot analyses demonstrated constitutive mRNA expression of MIP-1 gamma, MIP-1 alpha, C10, and IL-1 beta in both 4F7+ DC and LC. Lipopolysaccharide (LPS) treatment resulted in the upregulation of mRNA expression of all four cytokines and in a newly detected signal for TNF alpha. Immunoblot analysis showed constitutive secretion of MIP-1 gamma, with LPS treatment resulting in the upregulation of IL-1 beta production and in newly detected TNF alpha secretion. 4F7+ DC were also shown to express mRNA for the common gamma chain receptor of IL-2 and for the receptor of IL-4. Finally, we demonstrated freshly isolated 4F7+ DC to be equivalent to freshly isolated LC in their capacity to present alloantigen in the mixed leukocyte reaction (MLR) and to process and present purified protein derivative (PPD) to Th1 and Th2 clones. We conclude that 4F7 is a useful marker for positively sorting DC from dermis, spleen, and lymph nodes. Regardless of tissue source, 4F7+ DC exhibit uniform cytokine and antigen-presenting capacity profiles that mimic the properties of freshly isolated epidermal LC.
我们利用最近制备的4F7单克隆抗体,它能识别不同组织中树突状细胞(DC)上表达的一个表位,来新鲜分离并对这些细胞进行阳性分选,并与表皮朗格汉斯细胞(LC)比较,以表征它们的细胞因子模式和抗原呈递能力。逆转录聚合酶链反应(RT-PCR)和Northern印迹分析表明,MIP-1γ、MIP-1α、C10和IL-1β在4F7+ DC和LC中均有组成型mRNA表达。脂多糖(LPS)处理导致所有四种细胞因子的mRNA表达上调,并新检测到肿瘤坏死因子α(TNFα)的信号。免疫印迹分析显示MIP-1γ有组成型分泌,LPS处理导致IL-1β产生上调并新检测到TNFα分泌。4F7+ DC还显示表达IL-2共同γ链受体和IL-4受体的mRNA。最后,我们证明新鲜分离的4F7+ DC在混合淋巴细胞反应(MLR)中呈递同种异体抗原以及处理和呈递纯化蛋白衍生物(PPD)给Th1和Th2克隆的能力与新鲜分离的LC相当。我们得出结论,4F7是从真皮、脾脏和淋巴结中阳性分选DC的有用标志物。无论组织来源如何,4F7+ DC都表现出一致的细胞因子和抗原呈递能力谱,类似于新鲜分离的表皮LC的特性。