Crosstalk Between the Immune System and Plant-Derived Nanovesicles: A Study of Allergen Transporting

Abstract

Background: Nanometer-sized membrane-surrounded vesicles from different parts of plants including fruits are gaining increasing attention due to their anti-inflammatory and anticancer effects demonstrated by in vitro and in vivo studies, and as nanovectors for molecular delivery of exogenous substances. These nanomaterials are very complex and contain a diverse arsenal of bioactive molecules, such as nucleic acids, proteins, and lipids. Our knowledge about the transport of allergens in vesicles isolated from plant food is limited today.Methods: Here, to investigate the allergenicity of strawberry-derived microvesicles (MVs), nanovesicles (NVs), and subpopulations of NV, we have set up a multidisciplinary approach. The strategy combines proteomics-based protein identification, immunological investigations, bioinformatics, and data mining to gain biological insights useful to evaluate the presence of potential allergens and the immunoglobulin E (IgE) inhibitory activity of vesicle preparations.Results: Immunological test showed that several proteins of strawberry-derived vesicles compete for IgE binding with allergens spotted on the FABER biochip. This includes the known strawberry allergens Fra a 1, Fra a 3, and Fra a 4, and also other IgE-binding proteins not yet described as allergens in this food, such as gibberellin-regulated proteins, 2S albumin, pectate lyase, and trypsin inhibitors. Proteomics identified homologous sequences of the three strawberry allergens and their isoforms in total protein extract (TPE) but only Fra a 1 and Fra a 4 in the vesicle samples. Label-free quantitative proteomic analysis revealed no significant enrichment of these proteins in strawberry vesicles with respect to TPE.Conclusion: Immunological tests and bioinformatics analysis of proteomics data sets revealed that MVs and NVs isolated from strawberries can carry functional allergens their isoforms as well as proteins potentially allergenic based on their structural features. This should be considered when these new nanomaterials are used for human nutraceutical or biomedical applications.

Protein Biocargo and Anti-Inflammatory Effect of Tomato Fruit-Derived Nanovesicles Separated by Density Gradient Ultracentrifugation and Loaded with Curcumin

“…In recent years, plant-derived nanovesicles (PDNVs) have emerged due to their perspective towards biomedical and nanotechnological applications. They have been isolated from many homogenized plants and fruit juices, including tomato [ 1 , 2 ], citrus species [ 3 , 4 , 5 ], strawberry [ 6 , 7 ] and ginseng [ 8 , 9 ], just to mention some. In this paper, we use PDNVs as an umbrella term to refer to all kind of nano-sized particles, nanolipids, exosomes, nanovectors, etc.…”

Detection of Allergenic Proteins in Foodstuffs: Advantages of the Innovative Multiplex Allergen Microarray-Based Immunoassay Compared to Conventional Methods

“…Several mass-spectrometry-based methodologies and strategies can be used to identify proteins in food extracts [ 23 , 65 , 66 , 67 , 68 ]. The simultaneous identification and quantification of traces of allergenic proteins in complex mixtures such as processed foods can also be achieved [ 69 , 70 , 71 ].…”

“…More recently, the SPHIAa assay was applied to the multiplex allergen microarray FABER (Allergy Data Laboratories (ADL), Latina, Italy) [ 5 , 97 ] to obtain information on the allergens contained in food extracts. The SPHIAa method, combined with the FABER technology, represents a forefront tool exploiting a comprehensive panel of 244 allergens ( Table 2 ), namely 122 extracts and 122 purified molecules, including all the most important allergy markers, in addition to exclusive allergens not available in other test systems [ 5 , 23 , 68 ]. For instance, this method, associated with the use of appropriate sera containing the required IgE, was exploited to perform a comparative analysis of the allergen content in tomato exposed, or not exposed, to nickel (Ni) stress [ 101 ].…”

“…The same strategy combining immunological tests and bioinformatics analysis of proteomics data sets was applied to the analysis of allergens in three different strawberry-derived vesicle populations [ 68 ]. The application of the immunological method, namely the SPHIAa assay on the FABER system, showed that the vesicles carry all the three allergens so far described in strawberry, the Bet v 1-like protein Fra a 1, the LTP Fra a 3 and the profilin Fra a 4.…”

Plant-derived nanovesicles as an emerging platform for cancer therapy

“…A large amount of proteins, such as 3129 and 1018 proteins, are found in PDNVs from ginseng and citrus clementina fruit, respectively 59 , 98 . Nanovesicles isolated from strawberry contain more than 200 different proteins, among which heat shock proteins (HSPs) are often detected in PDNVs 99 . In addition, different types of HSPs are found in PDNVs like citrus fruit, grapefruit and tomato 100 , 101 , 102 .…”