Mielke V, Bauman J G, Sticherling M, Ibs T, Zomershoe A G, Seligmann K, Henneicke H H, Schröder J M, Sterry W, Christophers E
Department of Dermatology, University of Kiel, Federal Republic of Germany.
J Immunol. 1990 Jan 1;144(1):153-61.
A neutrophil-activating peptide (NAP)/IL-8 produced by LPS-stimulated human peripheral blood monocytes was biochemically purified and functionally characterized by different investigators. Work conducted in our laboratory showed that NAP/IL-8 as well as variants of this peptide are produced by a variety of cells (e.g., monocytes, T lymphocytes, endothelial cells) and that lesional psoriatic scales contain large amounts of biologically active NAP/IL-8. We now investigated human dermal fibroblasts for production of NAP/IL-8. The peptide was detected by immunocytochemistry by using the mAb 46E5. NAP/IL-8 mRNA was visualized by high resolutive fluorescent in situ hybridization with biotinylated antisense/sense RNA probes. Among the various stimuli used [human (h)rIL-1 alpha, hrTNF-alpha, hrIL-3, hr-granulocyte-macrophage-CSF, LPS, FMLP, and platelet-activating factor (PAF)] only hrIL-1 alpha (100 U/ml) and hrTNF-alpha (100 ng/ml) induced the transcription and translation of NAP/IL-8. In contrast to monocytes, LPS was without effect in cultured human dermal fibroblasts. Both NAP/IL-8 and NAP/IL-8 mRNA were found in the cytoplasm adjacent to the nucleus, but interestingly NAP/IL-8 mRNA was not restricted to the cytoplasm. In positive cells only two small bright spots were randomly distributed in the nucleus. Most likely these spots represent transcription sites where NAP/IL-8 mRNA is accumulated during gene expression. Our observations show that stimulation of dermal fibroblasts with the cytokines hrIL-1 alpha and hrTNF-alpha results in expression of IL-8.
由脂多糖刺激的人外周血单核细胞产生的一种中性粒细胞激活肽（NAP）/白细胞介素-8（IL-8）已被不同研究者进行了生化纯化和功能鉴定。我们实验室开展的研究表明，NAP/IL-8以及该肽的变体由多种细胞（如单核细胞、T淋巴细胞、内皮细胞）产生，并且银屑病皮损鳞屑中含有大量具有生物活性的NAP/IL-8。我们现在研究了人皮肤成纤维细胞产生NAP/IL-8的情况。使用单克隆抗体46E5通过免疫细胞化学检测该肽。通过用生物素化的反义/正义RNA探针进行高分辨率荧光原位杂交来观察NAP/IL-8 mRNA。在所使用的各种刺激物[人（h）重组白细胞介素-1α（hrIL-1α）、重组肿瘤坏死因子-α（hrTNF-α）、重组白细胞介素-3（hrIL-3）、重组粒细胞-巨噬细胞集落刺激因子（hr-粒细胞-巨噬细胞-CSF）、脂多糖、N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸（FMLP）和血小板活化因子（PAF）]中，只有hrIL-1α（100 U/ml）和hrTNF-α（100 ng/ml）诱导了NAP/IL-8的转录和翻译。与单核细胞不同，脂多糖对培养的人皮肤成纤维细胞没有作用。在紧邻细胞核的细胞质中发现了NAP/IL-8和NAP/IL-8 mRNA，但有趣的是，NAP/IL-8 mRNA并不局限于细胞质。在阳性细胞中，只有两个小亮点随机分布在细胞核中。这些亮点很可能代表转录位点，在基因表达过程中NAP/IL-8 mRNA在此处积累。我们观察到，用细胞因子hrIL-1α和hrTNF-α刺激皮肤成纤维细胞会导致IL-8的表达。
