Abos Gracia Beatriz, López Relaño Juan, Revilla Ana, Castro Lourdes, Villalba Mayte, Martín Adrados Beatriz, Regueiro Jose Ramon, Fernández-Malavé Edgar, Martínez Naves Eduardo, Gómez Del Moral Manuel
Department of Immunology, Faculty of Medicine, and 12 de Octubre Health Research Institute (imas12), Complutense University, Madrid, Spain.
Int Arch Allergy Immunol. 2017;173(1):12-22. doi: 10.1159/000467394. Epub 2017 May 10.
Allergic sensitization might be influenced by the lipids present in allergens, which can be recognized by natural killer T (NKT) cells on antigen-presenting cells (APCs). The aim of this study was to analyze the effect of olive pollen lipids in human APCs, including monocytes as well as monocyte-derived macrophages (Mϕ) and dendritic cells (DCs).
Lipids were extracted from olive (Olea europaea) pollen grains. Invariant (i)NKT cells, monocytes, Mϕ, and DCs were obtained from buffy coats of healthy blood donors, and their cell phenotype was determined by flow cytometry. iNKT cytotoxicity was measured using a lactate dehydrogenase assay. Gene expression of CD1A and CD1D was performed by RT-PCR, and the production of IL-6, IL-10, IL-12, and TNF-α cytokines by monocytes, Mϕ, and DCs was measured by ELISA.
Our results showed that monocytes and monocyte-derived Mϕ treated with olive pollen lipids strongly activate iNKT cells. We observed several phenotypic modifications in the APCs upon exposure to pollen-derived lipids. Both Mϕ and monocytes treated with olive pollen lipids showed an increase in CD1D gene expression, whereas upregulation of cell surface CD1d protein occurred only in Mϕ. Furthermore, DCs differentiated in the presence of human serum enhance their surface CD1d expression when exposed to olive pollen lipids. Finally, olive pollen lipids were able to stimulate the production of IL-6 but downregulated the production of lipopolysaccharide- induced IL-10 by Mϕ.
Olive pollen lipids alter the phenotype of monocytes, Mϕ, and DCs, resulting in the activation of NKT cells, which have the potential to influence allergic immune responses.
变应原中的脂质可能会影响变应性致敏作用,抗原呈递细胞(APC)上的自然杀伤T(NKT)细胞能够识别这些脂质。本研究的目的是分析橄榄花粉脂质对人APC的影响,包括单核细胞以及单核细胞衍生的巨噬细胞(Mϕ)和树突状细胞(DC)。
从油橄榄(Olea europaea)花粉粒中提取脂质。从健康献血者的血沉棕黄层中获取不变(i)NKT细胞、单核细胞、Mϕ和DC,并通过流式细胞术确定其细胞表型。使用乳酸脱氢酶测定法测量iNKT细胞毒性。通过逆转录聚合酶链反应(RT-PCR)检测CD1A和CD1D的基因表达,并通过酶联免疫吸附测定法(ELISA)测量单核细胞、Mϕ和DC产生白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、白细胞介素-12(IL-12)和肿瘤坏死因子-α(TNF-α)细胞因子的情况。
我们的结果表明,用橄榄花粉脂质处理的单核细胞和单核细胞衍生的Mϕ强烈激活iNKT细胞。我们观察到APC在暴露于花粉衍生脂质后出现了几种表型改变。用橄榄花粉脂质处理的Mϕ和单核细胞均显示CD1D基因表达增加,而细胞表面CD1d蛋白的上调仅发生在Mϕ中。此外,在人血清存在下分化的DC在暴露于橄榄花粉脂质时会增强其表面CD1d表达。最后,橄榄花粉脂质能够刺激IL-6的产生,但下调Mϕ产生的脂多糖诱导的IL-10。
橄榄花粉脂质改变单核细胞、Mϕ和DC的表型,导致NKT细胞活化,这有可能影响变应性免疫反应。