Specific induction of interleukin‐4‐producing cells in response to <i>in vitro</i> allergen stimulation in atopic individuals

Gabrielsson, Susanne; Paulie, Staffan; Rak, S.; Lagging, Eva; Hage, Marianne van; Härfast, B.

doi:10.1046/j.1365-2222.1997.560878.x

Cited by 31 publications

(39 citation statements)

References 38 publications

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“…All of them had high levels of peanut specific IgE antibodies. This corroborates studies demonstrating a significant positive correlation between allergen-specific serum IgE levels and high numbers of IL-4-producing cells [25] or active proliferation of T cells and Th2 cytokine production [26]. However, it is worth noting that peanut-specific T-helper cells in PBMC of peanut allergic patients were found to be only 0.6% of the total CD4+ T cells [27], thus explaining the quite low rate of responders in our ex vivo stimulation assays.…”

Section: Discussionsupporting

confidence: 91%

Effect of Heating and Glycation on the Allergenicity of 2S Albumins (Ara h 2/6) from Peanut

Vissers

Blanc

Skov³

et al. 2011

PLoS ONE

113

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BackgroundPeanut allergy is one of the most common and severe food allergies, and processing is known to influence the allergenicity of peanut proteins. We aimed to establish the effect of heating and glycation on the IgE-binding properties and biological activity of 2S albumins (Ara h 2/6) from peanut.Methodology/Principal FindingsNative Ara h 2/6 was purified from raw peanuts and heated in solution (15 min, 110°C) in the presence or absence of glucose. Ara h 2 and 6 were also purified from roasted peanut. Using PBMC and sera from peanut-allergic patients, the cellular proliferative potency and IgE reactivity (reverse EAST inhibition) and functionality (basophil degranulation capacity) of allergens were assessed. Heating Ara h 2/6 at 110°C resulted in extensive denaturation, hydrolysis and aggregation of the protein, whilst Ara h 2 and 6 isolated from roasted peanut retained its native conformation. Allergen stimulation of PBMC induced proliferation and Th2 cytokine secretion which was unaffected by thermal processing. Conversely, IgE reactivity and functionality of Ara h 2/6 was decreased by heating. Whilst heating-glycation further reduced the IgE binding capacity of the proteins, it moderated their loss of histamine releasing capacity. Ara h 2 and 6 purified from roasted peanut demonstrated the same IgE reactivity as unheated, native Ara h 2/6.Conclusions/SignificanceAlthough no effect of processing on T-cell reactivity was observed, heat induced denaturation reduced the IgE reactivity and subsequent functionality of Ara h 2/6. Conversely, Ara h 2 and 6 purified from roasted peanut retained the structure and IgE reactivity/functionality of the native protein which may explain the allergenic potency of this protein. Through detailed molecular study and allergenicity assessment approaches, this work then gives new insights into the effect of thermal processing on structure/allergenicity of peanut proteins.

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Section: Discussionsupporting

confidence: 91%

Effect of Heating and Glycation on the Allergenicity of 2S Albumins (Ara h 2/6) from Peanut

Vissers

Blanc

Skov³

et al. 2011

PLoS ONE

113

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“…The unaltered production levels of IL-5 and IL-13 (cytokines representing Th2-pathway) are consistent with a slower reacting T-cell compartment in these patients [19]. The frequency of allergen-specific T-cells will still be significant, probably in the order of 1 : 300, in these patients as they are in the pollen season and this arm of the immune system will still be triggered in vivo which compromises the Th2 analysis ex vivo [17, 20–22]. The monocyte compartment, being an essential part of the innate immune system, by definition will react faster on induced changes in the patient than the adaptive immune system, which is dependent on the frequency of allergen-specific T-cells and allergen-specific IgE antibody-forming B cells.…”

Section: Discussionmentioning

confidence: 80%

Citrus/Cydonia Compositum Subcutaneous Injections versus Nasal Spray for Seasonal Allergic Rhinitis: A Randomized Controlled Trial on Efficacy and Safety

Baars

Jong

Nierop³

et al. 2011

ISRN Allergy

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Background. Clinical experiences in vitro and clinical studies have demonstrated the curative potency and safety of Citrus/Cydonia compositum in seasonal allergic rhinitis treatment. Objectives. To compare the efficacy and safety of two routes of administration (nasal spray versus subcutaneous injections). Methodology: Design. a national, randomised, comparative clinical trial with two parallel groups. Participants. 23 patients fulfilled the study requirements. Intervention. after a one- or two-week wash-out period, 23 patients were randomized, to a 6-week treatment period. Outcomes. immunological and symptom severity changes and safety. Immunologic outcome assessments were blinded to group assignment. 23 patients were randomized and from 22/23 patients (11 in each group) blood samples were analyzed before and after treatment. Conclusion. Both routes of administration demonstrate immunological and clinical effects, with larger inflammatory and innate immunological effects of the nasal spray route and larger allergen-specific clinical effects of the subcutaneous route, and are safe.

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“…The viability was greater than 90% for all samples assayed. The cells were then pre-stimulated for 4 h in round-bottomed 5 ml polystyrene tubes at 10 6 cells/ml of TCM with or without allergen or phytohaemagglutinin (PHA) in concentrations previously shown to induce IL-4 and IFN-γ production [15]. The allergens used were Aquagen SQ cat extract (8 kU/ml, ALK-Abelló, Hørsholm, Denmark) or sterile filtered ovalbumin (10 µ g/ml, SigmaAldrich Sweden AB, Stockholm, Sweden).…”

Section: Blood/plasma Collection and Preparation Of Mononuclear Cellsmentioning

confidence: 99%

“…The reverse ELISpot assay, mainly as described by Gabrielsson and colleagues [15], was used for quantification of the numbers of cytokine-secreting cells. Briefly, 100 µ l of coating antibodies was added to 96-well nitrocellulose plates (Millipore Corp., Bedford, MA, USA) at 10 µ g/ml in sterile filtered carbonate buffer (pH 9·6).…”

Section: Quantification Of Cytokine Secretionmentioning

confidence: 99%

“…An allergen-specific IgE level ≥ 0·35 kU/l was considered positive. (12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25) Total IgE was measured in plasma by the ImmunoCAP® IgE fluorescence enzyme immunoassay (FEIA) method (Pharmacia Diagnostics AB, detection limit 2·0 kU/l) according to the manufacturer's instructions.…”

Section: Allergen-specific Ige Determinationmentioning

confidence: 99%

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Relationship between maternal and child cytokine responses to allergen and phytohaemagglutinin 2 years after delivery

Larsson

Nilsson

Höglind

et al. 2006

Clinical and Experimental Immunology

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SummaryLittle is known at present about the relation between parental and child cytokine profiles. In this study we aimed to investigate the cytokine profile of 2-year-old children and their corresponding mothers, 2 years after delivery. Peripheral blood mononuclear cells (PBMC) were isolated from IgEsensitized ( n = = = = 15) and non-sensitized ( n = = = = 58) 2-year-old children and their mothers. The responses to ovalbumin, cat and phytohaemagglutinin (PHA) were investigated using the enzyme-linked immunospot (ELISpot) technique. Interferon (IFN)-γ γ γ γ -, interleukin (IL)-4-, IL-10-and IL-12-producing cells were enumerated. At 2 years of age, IgE-sensitized children exhibited increased numbers of IL-4-producing cells in response to PHA and also showed an increase in IL-10-and IL-12-producing cells to allergen that was more pronounced in response to ovalbumin than to cat. A statistically significant increase in the numbers of IFN-γ γ γ γ -, IL-10-and IL-12-producing cells to the allergens, but not to PHA, was found in the mothers of IgE-sensitized children irrespective of their own atopic status. IgE levels and cytokine responses were correlated between the mothers and their children, indicating that cytokine responses to both allergen and PHA might be governed by genetic factors. We speculate that the increased cytokine response to allergen, as opposed to the allergic status of the mother, might be a better predictor and/or a risk factor for the child to develop IgE-sensitization in early life.

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Specific induction of interleukin‐4‐producing cells in response to in vitro allergen stimulation in atopic individuals

Abstract: Allergen-specific IL-4 producing cells in a peripheral blood mononuclear cell (PBMC) culture can be detected by ELISPOT and the response can synergistically be enhanced by suboptimal concentrations of PHA.

Cited by 31 publications

References 38 publications

Effect of Heating and Glycation on the Allergenicity of 2S Albumins (Ara h 2/6) from Peanut

Effect of Heating and Glycation on the Allergenicity of 2S Albumins (Ara h 2/6) from Peanut

Citrus/Cydonia Compositum Subcutaneous Injections versus Nasal Spray for Seasonal Allergic Rhinitis: A Randomized Controlled Trial on Efficacy and Safety

Relationship between maternal and child cytokine responses to allergen and phytohaemagglutinin 2 years after delivery

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