Torii H, Hosoi J, Beissert S, Xu S, Fox F E, Asahina A, Takashima A, Rook A H, Granstein R D
Massachusetts General Hospital/Harvard Cutaneous Biology Research Center, Boston, USA.
J Leukoc Biol. 1997 Feb;61(2):216-23. doi: 10.1002/jlb.61.2.216.
Calcitonin gene-related peptide (CGRP) inhibits antigen presentation by Langerhans cells (LC) and macrophages, and LC are anatomically associated with CGRP-containing epidermal nerves. To determine whether CGRP may produce some of its functional effects through regulation of cytokine expression, we utilized enzyme-linked immunosorbent assay (ELISA) of conditioned supernatants to examine production of interleukin (IL)-10 and IL-1 beta protein in the LC-like cell line XS52 as well as the reverse transcriptase-polymerase chain reaction (RT-PCR) to examine levels of mRNA for IL-10, IL-1 beta, and the 40-kDa subunit (p40) of IL-12. CGRP augmented the lipopolysaccharide (LPS) and granulocyte-macrophage colony-stimulating factor (GM-CSF) -induced release of IL-10 protein and the induced expression of IL-10 mRNA in these cells. However, it suppressed the induction of release of IL-1 beta protein and the induction of mRNA for IL-12 p40 and IL-1 beta by LPS and GM-CSF. Regulation of cytokine expression in peritoneal macrophages was also examined. By ELISA, the LPS-induced expression of IL-10 was augmented by CGRP, whereas the induction of IL-1 beta was suppressed. Northern analysis demonstrated augmentation of LPS-induced IL-10 mRNA levels and inhibition of LPS-induced IL-1 beta mRNA by CGRP. CGRP inhibited the LPS-induced induction of IL-12 mRNA as assessed by RT-PCR. Up-regulation of B7-2 expression by LPS and GM-CSF was suppressed by CGRP in both XS52 cells and macrophages, as previously reported. This suppression, however, could be abrogated by co-culture with neutralizing antibodies to IL-10. Furthermore, the presence of neutralizing antibodies to IL-10 during exposure of epidermal cells (EC) to CGRP prevented the CGRP-mediated suppression of EC presentation of tumor-associated antigens (from the S1509a spindle cell carcinoma) for elicitation of delayed-type hypersensitivity in S1509a-immune mice. These data suggest that suppression of antigen-presenting function by CGRP is mediated, at least in part, by changes in cytokine expression that favor less robust antigen presentation for cell-mediated immunity.
降钙素基因相关肽（CGRP）可抑制朗格汉斯细胞（LC）和巨噬细胞的抗原呈递，且LC在解剖学上与含CGRP的表皮神经相关。为确定CGRP是否可能通过调节细胞因子表达产生其部分功能效应，我们利用条件培养基的酶联免疫吸附测定（ELISA）检测了LC样细胞系XS52中白细胞介素（IL）-10和IL-1β蛋白的产生，并利用逆转录聚合酶链反应（RT-PCR）检测了IL-10、IL-1β及IL-12的40 kDa亚基（p40）的mRNA水平。CGRP增强了脂多糖（LPS）和粒细胞巨噬细胞集落刺激因子（GM-CSF）诱导的这些细胞中IL-10蛋白的释放及IL-10 mRNA的诱导表达。然而，它抑制了LPS和GM-CSF诱导的IL-1β蛋白释放及IL-12 p40和IL-1β mRNA的诱导。我们还检测了CGRP对腹腔巨噬细胞中细胞因子表达的调节。通过ELISA检测，CGRP增强了LPS诱导的IL-10表达，而抑制了IL-1β的诱导表达。Northern分析表明，CGRP增强了LPS诱导的IL-10 mRNA水平，并抑制了LPS诱导的IL-1β mRNA。通过RT-PCR评估，CGRP抑制了LPS诱导的IL-12 mRNA的诱导。如先前报道，在XS52细胞和巨噬细胞中，CGRP均抑制了LPS和GM-CSF诱导的B7-2表达上调。然而，与抗IL-10中和抗体共培养可消除这种抑制作用。此外，在表皮细胞（EC）暴露于CGRP期间存在抗IL-10中和抗体，可阻止CGRP介导的EC对肿瘤相关抗原（来自S1509a梭形细胞癌）呈递的抑制，从而在S1509a免疫小鼠中引发迟发型超敏反应。这些数据表明，CGRP对抗抗原呈递功能的抑制至少部分是由细胞因子表达的变化介导的，这些变化有利于细胞介导免疫中较弱的抗原呈递。
