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Hydrophobicity drives receptor-mediated uptake of heat-processed proteins by THP-1 macrophages and dendritic cells, but not cytokine responses
bioRxiv - Immunology Pub Date : 2020-07-02 , DOI: 10.1101/2020.07.02.184317
Ying Deng , Coen Govers , Malgorzata Teodorowicz , Ieva Liobyte , Ilaria De Simone , Kasper Hettinga , Harry J. Wichers

Impact of processing on immunogenicity of food proteins has clearly been demonstrated, but the underlying mechanisms are still unclear. In our previous study, the uptake of the cow’s milk protein β-lactoglobulin (BLG) by THP-1 macrophages varied after applying different processing methods and was positively correlated with hydrophobicity and aggregation. Here we applied the same 3 processing methods: wet heating (60 °C) and low- or high-temperature (50 °C or 130 °C, respectively) dry heating in absence or presence of reducing sugars (i.e. glucose, lactose or galacto-oligosaccharide) to lysozyme and thyroglobulin, which have different pI or molecular weight compared to BLG, respectively. Uptake of the soluble fraction was tested in two types of, genetically homogeneous, antigen-presenting cells (macrophages and dendritic cells derived from THP-1 monocytes). This revealed a strong correlation between the uptake of the different protein samples by macrophages and dendritic cells, and confirmed the key role of hydrophobicity, over aggregation, in determining the uptake. Several uptake routes were shown to contribute to the uptake of BLG by macrophages. However, cytokine responses following exposure of macrophages to BLG samples were not related to the levels of uptake. Heat-treatment-mediated increases in uptake did thus not induce a response in our read-out systems. Together, our results demonstrate that heat-treatment-induced increased hydrophobicity is the prime driving factor in uptake, but not in cytokine production, by THP-1 macrophages.

中文翻译:

疏水性通过THP-1巨噬细胞和树突状细胞驱动受体介导的热加工蛋白的摄取,而不是细胞因子的响应

加工过程对食物蛋白免疫原性的影响已得到明确证明,但其潜在机制仍不清楚。在我们先前的研究中,采用不同的加工方法后,THP-1巨噬细胞对牛乳蛋白β-乳球蛋白(BLG)的摄取有所不同,并且与疏水性和聚集呈正相关。在这里,我们采用了相同的3种处理方法:湿加热(60°C)和低温或高温(分别为50°C或130°C)在没有或没有还原糖(例如葡萄糖,乳糖或半乳糖)的情况下进行干加热-寡糖)至溶菌酶和甲状腺球蛋白,与BLG相比分别具有不同的pI或分子量。在两种类型的,遗传上均一的,抗原呈递细胞(来自THP-1单核细胞的巨噬细胞和树突状细胞)。这揭示了巨噬细胞和树突状细胞对不同蛋白质样品的摄取之间的强相关性,并证实了疏水性(超过聚集)在决定摄取中的关键作用。已显示几种摄取途径有助于巨噬细胞摄取BLG。但是,巨噬细胞暴露于BLG样品后的细胞因子反应与摄取水平无关。因此,热处理介导的摄取增加并未在我们的读出系统中引起反应。总之,我们的结果表明,热处理诱导的疏水性增加是THP-1巨噬细胞摄取而不是细胞因子产生的主要驱动因素。这揭示了巨噬细胞和树突状细胞对不同蛋白质样品的摄取之间的强相关性,并证实了疏水性(超过聚集)在决定摄取中的关键作用。已显示几种摄取途径有助于巨噬细胞摄取BLG。但是,巨噬细胞暴露于BLG样品后的细胞因子反应与摄取水平无关。因此,热处理介导的摄取增加并未在我们的读出系统中引起反应。总之,我们的结果表明,热处理诱导的疏水性增加是THP-1巨噬细胞摄取而不是细胞因子产生的主要驱动因素。这揭示了巨噬细胞和树突状细胞对不同蛋白质样品的摄取之间的强相关性,并证实了疏水性(超过聚集)在决定摄取中的关键作用。已显示几种摄取途径有助于巨噬细胞摄取BLG。但是,巨噬细胞暴露于BLG样品后的细胞因子反应与摄取水平无关。因此,热处理介导的摄取增加并未在我们的读出系统中引起反应。总之,我们的结果表明,热处理诱导的疏水性增加是THP-1巨噬细胞摄取而不是细胞因子产生的主要驱动因素。已显示几种摄取途径有助于巨噬细胞摄取BLG。但是,巨噬细胞暴露于BLG样品后的细胞因子反应与摄取水平无关。因此,热处理介导的摄取增加并未在我们的读出系统中引起反应。总之,我们的结果表明,热处理诱导的疏水性增加是THP-1巨噬细胞摄取而不是细胞因子产生的主要驱动因素。已显示几种摄取途径有助于巨噬细胞摄取BLG。但是,巨噬细胞暴露于BLG样品后的细胞因子反应与摄取水平无关。因此,热处理介导的摄取增加并未在我们的读出系统中引起反应。总之,我们的结果表明,热处理诱导的疏水性增加是THP-1巨噬细胞摄取而不是细胞因子产生的主要驱动因素。
更新日期:2020-07-03
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