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Abstract：
Melittin, the major active peptide of honeybee venom (BV), has potential for use in adjuvant immunotherapy. The immune system response to different stimuli depends on the secretion of different metabolites from macrophages. One potent stimulus is lipopolysaccharide (LPS), a component isolated from gram-negative bacteria, which induces the secretion of pro-inflammatory cytokines in macrophage cell cultures. This secretion is amplified when LPS is combined with melittin. In the present study, pure melittin was isolated from whole BV by flash chromatography to obtain pure melittin. The ability of melittin to enhance the release of tumour necrosis factor-α (TNF-α), Interleukin (IL-1β, IL-6, and IL-10) cytokines from a macrophage cell line (THP-1) was then assessed. The response to melittin and LPS, applied alone or in combination, was characterised by metabolic profiling, and the metabolomics results were used to evaluate the potential of melittin as an immune adjuvant therapy. The addition of melittin enhanced the release of inflammatory cytokines induced by LPS. Effective chromatographic separation of metabolites was obtained by liquid chromatography-mass spectrometry (LC-MS) using a ZIC-pHILIC column and an ACE C4 column. The levels of 108 polar and non-polar metabolites were significantly changed (p ˂ 0.05) following cell activation by the combination of LPS and melittin when compared to untreated control cells. Overall, the findings of this study suggested that melittin might have a potential application as a vaccine adjuvant.
摘要：
蜂毒素,蜜蜂毒液（BV）的主要活性肽，具有用于辅助免疫疗法的潜力。免疫系统对不同刺激的反应取决于巨噬细胞分泌不同的代谢物。一种有效的刺激是脂多糖（LPS），从革兰氏阴性细菌中分离出的成分，在巨噬细胞培养物中诱导促炎细胞因子的分泌。当LPS与蜂毒素组合时,这种分泌被放大。在本研究中,通过快速色谱法从整个BV中分离纯的蜂毒素以获得纯的蜂毒素。蜂毒肽增强肿瘤坏死因子-α（TNF-α）释放的能力，白细胞介素（IL-1β，然后评估来自巨噬细胞系(THP-1)的IL-6和IL-10)细胞因子。对蜂毒肽和LPS的反应，单独或组合应用，以代谢谱为特征，代谢组学结果用于评估蜂毒素作为免疫辅助治疗的潜力。蜂毒素的添加增强了LPS诱导的炎性细胞因子的释放。使用ZIC-pHILIC柱和ACEC4柱通过液相色谱-质谱（LC-MS）获得了代谢物的有效色谱分离。当与未处理的对照细胞相比时,在通过LPS和蜂毒素的组合的细胞活化后,108个极性和非极性代谢物的水平显著改变(P<0.05)。总的来说,这项研究的结果表明，蜂毒素可能具有作为疫苗佐剂的潜在应用。
