Formanek M, Knerer B, Temmel A, Thurnher D, Millesi W, Kornfehl J
Department of Otorhinolaryngology, Head and Neck Surgery, University of Vienna Medical School, Austria.
J Oral Pathol Med. 1998 May;27(5):202-6. doi: 10.1111/j.1600-0714.1998.tb01942.x.
The aim of the present study is to examine the cytokine expression and corresponding receptor pattern of human oral mucosa-derived keratinocytes. The mRNA expression of these cytokines from isolated and purified cells was measured by a RT-PCR method, the protein production by ELISA, and the receptor expression was determined by FACS analysis. In freshly isolated oral keratinocytes, IL-1alpha, IL-1alpha receptor antagonist, IL-6, IL-8, TGF-beta, TNF-alpha, and bFGF were detectable at the protein and mRNA level, whereas PDGF and TGF-alpha were found only at the mRNA level. There were no detectable signals for IL-2 and IL-4. The cytokine production at the protein level was independent from stimulation with PMA (phorbol myristate acetate). Unstimulated commercially available and primary isolated epidermal keratinocytes showed similar cytokine pattern except a lack of IL-6. FACS analysis revealed receptor expression on oral keratinocytes for IL-1, IL-2, IL-4, IL-6, EGF, IFN-gamma, and PDGF. In addition, receptor mRNA for IL-8, TNF-alpha, FGF-2 and KGF could be detected, but not for IL-10. Our results show that human oral keratinocytes produce a cytokine panel comparable to epidermal keratinocytes. In contrast to the epidermis, IL-6 was produced by human oral keratinocytes constitutively without prior stimulation, which may indicate their active regulation role in the maintenance of the oral mucosa.
本研究的目的是检测人口腔黏膜来源角质形成细胞的细胞因子表达及相应的受体模式。通过逆转录聚合酶链反应(RT-PCR)方法检测分离纯化细胞中这些细胞因子的mRNA表达,采用酶联免疫吸附测定(ELISA)法检测蛋白生成情况,并用荧光激活细胞分选(FACS)分析确定受体表达。在新鲜分离的口腔角质形成细胞中,可在蛋白和mRNA水平检测到白细胞介素-1α(IL-1α)、IL-1α受体拮抗剂、IL-6、IL-8、转化生长因子-β(TGF-β)、肿瘤坏死因子-α(TNF-α)和碱性成纤维细胞生长因子(bFGF),而血小板衍生生长因子(PDGF)和TGF-α仅在mRNA水平被发现。未检测到IL-2和IL-4的信号。蛋白水平的细胞因子生成与佛波酯(PMA)刺激无关。未受刺激的市售及原代分离的表皮角质形成细胞显示出相似的细胞因子模式,但缺乏IL-6。FACS分析显示口腔角质形成细胞上存在IL-1、IL-2、IL-4、IL-6、表皮生长因子(EGF)、γ干扰素(IFN-γ)和PDGF的受体。此外,可检测到IL-8、TNF-α、成纤维细胞生长因子-2(FGF-2)和角质形成细胞生长因子(KGF)的受体mRNA,但未检测到IL-10的受体mRNA。我们的结果表明,人口腔角质形成细胞产生的细胞因子谱与表皮角质形成细胞相当。与表皮不同,人口腔角质形成细胞在未预先刺激的情况下可组成性产生IL-6,这可能表明它们在维持口腔黏膜方面具有积极的调节作用。