Supplementary MaterialsS1 Fig: There is no significant difference between your first uptake value of BLG, lysozyme and thyroglobulin for THP-1 derived macrophages (M?, A) and immature dendritic cells (iDC, B). of thyroglobulin was prominent upon high-temperature and wet-heating dry-heating in the current presence of GOS. Thyroglobulin (THY) was neglected or warmed (W, H or L) in the lack or existence of saccharides (glu, lac or GOS) and aggregation in the soluble small fraction was assessed using size exclusion chromatography. The info points represent the common ideals of 2 3rd party sample models.(PDF) pone.0236212.s003.pdf (201K) GUID:?C05739E8-A606-45E3-B735-C7ECF6C71169 S4 Fig: Significant physicochemical modifications of indigenous lysozyme were mainly induced by high-temperature dry-heating. Lysozyme (LYS) was neglected or warmed (W, H or L) in the lack or existence of saccharides (glu, lac or GOS) and several physicochemical guidelines (we.e., solubility, loss of amino group, AGE-related fluorescence, hydrophobicity, -helix and -sheet structure) were measured. Calyculin A The results represent the mean values SD of n = 4 measurements of 2 impartial experiments based on 2 impartial sample sets. Statistical differences compared to native LYS were calculated with Dunnetts Test: *p 0.05; **p 0.01; ***p 0.001.(PDF) pone.0236212.s004.pdf (222K) GUID:?40BE293D-43ED-43E0-BF53-D6B728D7B100 S5 Fig: Wet-heated lysozyme samples had physicochemical properties that are similar to native lysozyme. Lysozyme (LYS) was untreated or heated (W, H or L) in the absence or presence of saccharides (glu, lac or GOS) and tested for solubility, uptake by THP-1 macrophages (uptake), AGE formation (AGE), glycation (OPA), percentage of -helix (helix) or -sheet (sheet), and exposure of hydrophobic regions (ANS). The aggregation-related parameters, proportion of monomer, oligomers and polymers are not shown as they did not differ between the samples.(PDF) pone.0236212.s005.pdf (26K) GUID:?40EEA8B8-4467-4F8D-97CB-6BEFB73FD09C S1 Table: LPS contamination in samples. (PDF) pone.0236212.s006.pdf (417K) GUID:?5AFFDE9C-8589-4DF9-941E-9057FB4C7F83 S1 Protocol: Lipopolysaccharide (LPS) detection. (PDF) pone.0236212.s007.pdf (381K) GUID:?3E54E525-D2EA-4C85-9AD4-5568CBA8F414 Data Availability StatementAll relevant data are within the paper and Calyculin A its Supporting Information files. Abstract Although an impact of processing on immunogenicity of food proteins has clearly been demonstrated, the underlying mechanisms are still unclear. We applied 3 different 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, to -lactoglobulin (BLG), lysozyme and thyroglobulin, which represent dietary proteins with different pI or molecular weight. Uptake of the soluble fraction of the samples 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. Rabbit Polyclonal to DNAI2 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. Together, our results demonstrate that heat-treatment-induced increased hydrophobicity is the primary driving factor in uptake, however, not in cytokine creation, by THP-1 macrophages. Launch Eating proteins play a significant physiological role, not only in providing amino acids and energy, but also for the maturation and regulation of the immune system. Proteins are known to be involved in the regulation of chronic inflammation and be the cause of allergies [1]. For example, a Calyculin A higher intake of animal protein was found to enhance the pro-inflammatory response of macrophages in mice [2]. Most of the proteins that humans ingest have been through a heating process as part of the food processing, as a result of which structural modifications, such as glycation (with reducing sugar) and/or aggregation of proteins, may occur. Different processing methods would alter the immunogenicity of food protein in different manner. For example, the antigenicity of prawn allergen was found to be significantly increased after frying but decreased after boiling, acid treatment or high-pressure handling [3]. The allergenicity of dairy proteins was reported to diminish after glycation [4]. Within an previous research on -lactoglobulin (BLG), the main whey proteins of dairy [5], we noticed that heat handling under different circumstances changed the physicochemical properties and its own uptake by THP-1 macrophages. Specifically, heating system in option at 60 C for 3 times introduced adjustments in hydrophobicity and molecular fat, which were been shown to be the main element determinants for uptake. This event may be relevant for the.