Sunasee Rajesh, Araoye Erinolaoluwa, Pyram Dejhy, Hemraz Usha D, Boluk Yaman, Ckless Karina
Department of Chemistry, State University of New York at Plattsburgh, Hudson Hall, 101 Broad Street, Plattsburgh, NY, USA.
National Research Council of Canada, Montreal, Quebec, Canada.
Biochem Biophys Rep. 2015 Aug 14;4:1-9. doi: 10.1016/j.bbrep.2015.08.008. eCollection 2015 Dec.
Crystalline cellulose nanocrystals (CNCs) have emerged as novel materials for a wide variety of important applications such as nanofillers, nanocomposites, surface coatings, regenerative medicine and potential drug delivery. CNCs have a needle-like structure with sizes in the range of 100-200 nm long and 5-20 nm wide and a mean aspect ratio 10-100. Despite the great potential applicability of CNCs, very little is known about their potential immunogenicity. Needle-like materials have been known to evoke an immune response in particular to activate the (NOD-like receptor, pyrin domain-containing 3)-inflammasome/IL-1β (Interleukin 1β) pathway. In this study we evaluated the capacity of unmodified CNC and its cationic derivatives CNC-AEM (aminoethylmethacrylate)1, CNC-AEM2, CNC-AEMA(aminoethylmethacrylamide)1 and CNC-AEMA2 to stimulate NLRP3-inflammasome/IL-1β pathway and enhance the production of mitochondrial reactive oxygen species (ROS). Mouse macrophage cell line (J774A.1) was stimulated for 24 h with 50 µg/mL with unmodified CNC and its cationic derivatives. Alternatively, J774A1 or PBMCs (peripheral blood mononuclear cells) were stimulated with CNC-AEMA2 in presence or absence of LPS (lipopolysaccharide). IL-1β secretion was analyzed by ELISA, mitochondrial function by JC-1 staining and ATP content. Intracellular and mitochondrial reactive oxygen species (ROS) were assessed by DCF-DA (2',7'-dichlorodihydrofluorescein diacetate) and MitoSOX, respectively. Mitochondrial ROS and extracellular ATP were significantly increased in cells treated with CNC-AEMA2, which correlates with the strongest effects on IL-1β secretion in non-primed cells. CNC-AEMA2 also induced IL-1βsecretion in LPS-primed and non-primed PBMCs. Our data suggest that the increases in mitochondrial ROS and ATP release induced by CNC-AEMA2 may be associated with its capability to evoke immune response. We demonstrate the first evidence that newly synthesized cationic cellulose nanocrystal derivative, CNC-AEMA2, has immunogenic properties, which may lead to the development of a potential non-toxic and safe nanomaterial to be used as a novel adjuvant for vaccines.
结晶纤维素纳米晶体（CNCs）已成为用于多种重要应用的新型材料，如纳米填料、纳米复合材料、表面涂层、再生医学和潜在的药物递送。CNCs具有针状结构，长度在100 - 200纳米范围内，宽度为5 - 20纳米，平均长径比为10 - 100。尽管CNCs具有巨大的潜在适用性，但关于它们潜在的免疫原性却知之甚少。已知针状材料会引发免疫反应，特别是激活含NOD样受体、含pyrin结构域3的炎性小体/白细胞介素1β（IL-1β）途径。在本研究中，我们评估了未修饰的CNC及其阳离子衍生物CNC - AEM（甲基丙烯酸氨基乙酯）1、CNC - AEM2、CNC - AEMA（甲基丙烯酰胺基乙胺）1和CNC - AEMA2刺激NLRP3炎性小体/IL-1β途径以及增强线粒体活性氧（ROS）产生的能力。用50μg/mL未修饰的CNC及其阳离子衍生物刺激小鼠巨噬细胞系（J774A.1）24小时。或者，在有或没有脂多糖（LPS）存在的情况下，用CNC - AEMA2刺激J774A1或外周血单核细胞（PBMCs）。通过ELISA分析IL-1β分泌，通过JC-1染色分析线粒体功能和ATP含量。分别用2',7'-二氯二氢荧光素二乙酸酯（DCF-DA）和MitoSOX评估细胞内和线粒体内的活性氧（ROS）。用CNC - AEMA2处理的细胞中线粒体ROS和细胞外ATP显著增加，这与对未致敏细胞中IL-1β分泌的最强作用相关。CNC - AEMA2还在LPS致敏和未致敏的PBMCs中诱导IL-1β分泌。我们的数据表明，CNC - AEMA2诱导的线粒体ROS增加和ATP释放可能与其引发免疫反应的能力有关。我们首次证明新合成的阳离子纤维素纳米晶体衍生物CNC - AEMA2具有免疫原性，这可能导致开发一种潜在的无毒且安全的纳米材料用作新型疫苗佐剂。
