Bruchhage Karl-Ludwig, Heinrichs Sabrina, Wollenberg Barbara, Pries Ralph
Department of Otorhinolaryngology, Head and Neck Surgery, University Clinic of Schleswig-Holstein Campus Lübeck, D-23538 Lübeck, Germany.
Oncol Lett. 2018 Mar;15(3):3985-3990. doi: 10.3892/ol.2018.7772. Epub 2018 Jan 11.
It has been shown that head and neck squamous cell carcinoma (HNSCC) are infiltrated by plasmacytoid dendritic cells (pDCs). The HNSCC TH2 biased microenvironment leads to strong alterations of the cellular functions of pDC and thus impairs the initiation and function of adequate immune responses. In this work we comprehensively analyzed the capacity of CpG-oligonucleotides to activate interferon (IFN)-α secretion of human pDC in the presence of HNSCC. IFN-α secretion was measured using the ELISA Technique. Class A CpG dinucleotide 2216 was used in different concentrations and time frames to stimulate the IFN-α production of human pDC from peripheral blood in the absence and presence of the HNSCC microenvironment. To elucidate single components that might induce the reduction of IFN-α secretion, pDC were exposed to different concentrations of HNSCC relevant cytokines such as IL-6, IL-8 and IL-10. In accordance to former experiments we found that HNSCC micro milieu severely depresses up to 75% of IFN-α secretion capacity of pDCs, if the stimulating Class A CpG 2216 is added to the culture. Preincubation of HNSCC supernatant leads to unrestorable reduction of IFN-α secretion in pDC and can not be restored by CpG 2216. Incubation of pDCs with single cytokines relevant for cancer progression within the HNSCC micro milieu show that IL-6 or IL-8 have no influence on the IFN-α secretion in pDCs, whereas IL-10 massively impairs the secretion in a dose dependent manner. This effect can be potentiated by synergistic incubation with IL-6 and can be abrogated by blocking antibodies to the IL-10 receptor. Interestingly, incubation with IL-10 is not the only factor that impairs the IFN-α secretion, as incubation with the whole HNSCC supernatant is even more effective in reducing the secretion, implying that additional factors play a role. We conclude that restoration of HNSCC induced TH2 bias could be improved by the inhibition of immune cell cytokine receptors in addition to immunostimulating approaches with CpG motifs.
已有研究表明，头颈部鳞状细胞癌（HNSCC）中有浆细胞样树突状细胞（pDC）浸润。HNSCC的TH2偏向性微环境导致pDC细胞功能发生强烈改变，从而损害了充分免疫反应的启动和功能。在这项研究中，我们全面分析了在HNSCC存在的情况下，CpG寡核苷酸激活人pDC分泌干扰素（IFN）-α的能力。使用酶联免疫吸附测定（ELISA）技术检测IFN-α的分泌。在不存在和存在HNSCC微环境的情况下，使用不同浓度和时间框架的A类CpG二核苷酸2216刺激外周血人pDC产生IFN-α。为了阐明可能导致IFN-α分泌减少的单一成分，将pDC暴露于不同浓度的与HNSCC相关的细胞因子，如白细胞介素（IL）-6、IL-8和IL-10。与先前的实验一致，我们发现，如果将刺激A类CpG 2216添加到培养物中，HNSCC微环境会严重抑制pDC高达75%的IFN-α分泌能力。HNSCC上清液的预孵育导致pDC中IFN-α分泌不可恢复的减少，且不能被CpG 2216恢复。在HNSCC微环境中，将pDC与与癌症进展相关的单一细胞因子孵育表明，IL-6或IL-8对pDC中的IFN-α分泌没有影响，而IL-10以剂量依赖性方式大量损害分泌。与IL-6协同孵育可增强这种效应，而抗IL-10受体的阻断抗体可消除这种效应。有趣的是，与IL-10孵育并不是损害IFN-α分泌的唯一因素，因为与整个HNSCC上清液孵育在减少分泌方面甚至更有效，这意味着还有其他因素起作用。我们得出结论，除了用CpG基序进行免疫刺激方法外，通过抑制免疫细胞细胞因子受体可以改善HNSCC诱导的TH2偏向的恢复。
