Evaluation of Immunoreactivity of Pea (<i>Pisum sativum</i>) Albumins in BALB/c and C57BL/6 Mice

Chudzik-Kozłowska, Justyna; Wasilewska, Ewa; Złotkowska, Dagmara

doi:10.1021/acs.jafc.0c00297

Cited by 14 publications

(14 citation statements)

References 47 publications

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“…Here, we describe monoclonal antibodies raised against selected pea globulins. Monoclonal antibodies have already been raised against pea albumin 1b, but this protein shows negligible allergenicity in contrast to other legume 2S albumins . Our antibodies against pea globulins showed no cross-reactivity to other legume proteins, allowing their integration into a multiplex legume protein assay for the simultaneous detection of the four most common legume-derived protein ingredients and potential allergens.…”

Section: Introductionmentioning

confidence: 87%

Development of Monoclonal Antibodies against Pea Globulins for Multiplex Assays Targeting Legume Proteins

Lidzba

Arteaga

Schiermeyer

et al. 2021

J. Agric. Food Chem.

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Legume proteins are widely used as food ingredients, but only some (soybean, lupin, and peanut) must be declared under consumer safety regulations to protect allergy sufferers. It is not yet mandatory to declare pea proteins as allergens even though they are predicted to be allergenic based on cross-reactivity in sensitized people. The processing of legume proteins can modify their allergenic properties and hence the need for specific and precise methods for the detection of all major legume allergens. There are many commercially available tests for known food allergens but not for ingredients that are yet to be classified as allergenic. We therefore generated sets of pea-specific antibodies targeting globulins to be used in a multiplex assay for the simultaneous detection of soybean, lupin, peanut, and pea proteins. We focused on the 7S globulin family, which is the least conserved among the four legumes, allowing the specific detection of proteins from each species. Having confirmed the specificity and sensitivity of the multiplex assay, we evaluated different processing steps for proteins rich in pea globulins to demonstrate the impact of food processing on antibody binding. Our sensitive multiplex assay provides a fast and reliable method for the specific detection of soybean, lupin, peanut, and pea allergens and is therefore ideal for food safety and authenticity testing applications.

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Section: Introductionmentioning

confidence: 87%

Development of Monoclonal Antibodies against Pea Globulins for Multiplex Assays Targeting Legume Proteins

Lidzba

Arteaga

Schiermeyer

et al. 2021

J. Agric. Food Chem.

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“…The crude barley extract (cBP, at a concentration of 1 mg/mL) and its fractions eluted by DEAE ion-exchange chromatography (15 μL/per lane) were subjected to SDS- PAGE according to the standard protocol used in the laboratory [ 15 ]. Briefly, the proteins were separated on gels (15% for cBP and 12.5% for fractions), under a constant current of 30 mA, using a Mini-PROTEAN system (Bio-Rad Laboratories, Inc., Warsaw, Poland).…”

Section: Methodsmentioning

confidence: 99%

The Immune System Response to 15-kDa Barley Protein: A Mouse Model Study

et al. 2022

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Barley (Hordeum vulgare L.) proteins are taxonomically homologous to wheat proteins and react with sera from patients with baker’s asthma. In the current work, the crude extract of barley proteins was divided into six fractions on DEAE-Sepharose. Their immunoreactivity in reacting with sera from patients with a confirmed food allergy varied, and the 15-kDa fraction (B–FrVI) showed the strongest response. In silico analysis confirmed that 15-kDa B-FrVI protein belongs to the trypsin/amylase inhibitor family and to a group of MHC type II allergens. In the next step, the immunogenicity of the B-FrVI was examined in a mouse model. It was shown that, compared to the PBS group, administration of B-FrVI to mice induced almost 2× higher amounts of specific IgG, ~217, and IgA ~29, as early as day 28 after immunization, regardless of the route (intraperitoneal or oral) of antigen administration (p < 0.0001). An ELISpot for B-cell responses confirmed it. Stimulation of mesenteric lymphocytes with pure B-FrVI significantly increased (p < 0.001) the proliferation of lymphocytes from all groups compared to cells growing in media only and stimulated with lyophilized beer. The experiments prove the strong immunogenicity of the 15-kDa B-FrVI protein and provide a basis for future studies of the allergenic nature of this protein.

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“…Blood was coagulated and centrifuged using the conditions described above. Samples were stored at a temperature of −20 °C until analysis [ 42 ].…”

Section: Methodsmentioning

confidence: 99%

“…The absorbance was measured at 405 nm on a Jupiter UVM spectrophotometer (ASYS-Hitech GmbH, Eugendorf, Austria). The endpoint titers (EPTs) were expressed as the reciprocal dilution of the last sample dilution of 0.1 OD above the negative control [ 42 , 43 , 44 , 45 ].…”

Section: Methodsmentioning

confidence: 99%

OVA-Experienced CD4+ T Cell Transfer and Chicken Protein Challenge Affect the Immune Response to OVA in a Murine Model

Fuc

Złotkowska

Wasilewska

et al. 2021

IJMS

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Chicken meat is often a major component of a modern diet. Allergy to chicken meat is relatively rare and occurs independently or in subjects allergic to ovalbumin (OVA). We examined the effect of adoptive transfer of OVA-CD4+ T cells on the immune response to OVA in mice fed chicken meat. Donor mice were injected intraperitoneally with 100 µg of OVA with Freund’s adjuvant two times over a week, and CD4+ T cells were isolated from them and transferred to naïve mice (CD4+/OVA/ChM group), which were then provoked with OVA with FA and fed freeze-dried chicken meat for 14 days. The mice injected with OVA and fed chicken meat (OVA/ChM group), and sensitized (OVA group) and healthy (PBS group) mice served as controls. Humoral and cellular response to OVA was monitored over the study. The CD4+/OVA/ChM group had lowered levels of anti-OVA IgG and IgA, and total IgE. There were significant differences in CD4+, CD4+CD25+, and CD4+CD25+Foxp3+ T cells between groups. OVA stimulation decreased the splenocyte proliferation index and IFN-γ secretion in the CD4+/OVA/ChM group compared to the OVA group. IL-4 was increased in the OVA/ChM mice, which confirms allergenic potential of the egg–meat protein combination. Transfer of OVA-experienced CD4+ T cells ameliorated the negative immune response to OVA.

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Evaluation of Immunoreactivity of Pea (Pisum sativum) Albumins in BALB/c and C57BL/6 Mice

Cited by 14 publications

References 47 publications

Development of Monoclonal Antibodies against Pea Globulins for Multiplex Assays Targeting Legume Proteins

Development of Monoclonal Antibodies against Pea Globulins for Multiplex Assays Targeting Legume Proteins

The Immune System Response to 15-kDa Barley Protein: A Mouse Model Study

OVA-Experienced CD4+ T Cell Transfer and Chicken Protein Challenge Affect the Immune Response to OVA in a Murine Model

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