<i>In vitro</i> detection of chemical allergens: an optimized assay using mouse bone marrow‐derived dendritic cells

Battais, F.; Huppert, Cécile; Langonné, Isabelle; Müller, S.; Sponne, Isabelle

doi:10.1111/cod.12829

Cited by 12 publications

(42 citation statements)

References 59 publications

(83 reference statements)

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“…To overcome this problem, granulocyte-macrophage colony-stimulating factor (GM-CSF)-differentiated bone marrow-derived DC (BMDC) culture methods have been developed to generate DCs in large numbers (5). The availability of cultured BMDCs facilitates their use in in vitro screening methods, including immunogenicity tests for vaccines and toxicity tests for allergens (6)(7)(8). In addition to well-characterized murine BMDC culture methods, such methods have been developed for veterinary species, including dogs [Ricklin (9)], cats (10), cattle (11), sheep (12), pigs (13), and chickens (14).…”

Section: Introductionmentioning

confidence: 99%

In vitro Chicken Bone Marrow-Derived Dendritic Cells Comprise Subsets at Different States of Maturation

et al. 2020

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Research in chickens has been fundamental for the discovery of basic aspects of the immune system and has led to an interest in the in-depth characterization of avian immune cell types including dendritic cells (DCs). The in vitro generation and expansion of chicken bone marrow-derived DCs (chBMDCs) in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) has provided a way to study chicken DCs, which are only present at limited cell numbers in vivo. This method has been employed to study the interactions between chicken DCs and pathogens or vaccines. However, a detailed characterization of the chBMDC culture is still lacking. In the present study, we performed an elaborate phenotypical and functional analysis of the chBMDC culture and addressed its heterogeneity. After 8 days of culture, chBMDCs comprised major histocompatibility complex class II (MHC-II) low and MHC-II high subsets with different morphologies. Compared with MHC-II low chBMDCs, the MHC-II high subset showed a more mature phenotype, with higher expressions of CD1.1, CD40, CD80, CCR7, and CD83, and a relatively low opsonophagocytic capacity. Nevertheless, MHC-II high chBMDCs did not show an increased capacity to induce T-cell proliferation. Therefore, MHC-II high chBMDCs were found to be semi-mature. Interestingly, the presence of the semi-mature MHC-II high chBMDC subset reduced when cells were cultured in the presence of IL-4. Finally, prolonged cell culture after fluorescence-activated cell sorting (FACS) converted the semi-mature MHC-II high subset back into the immature phenotype of the MHC-II low subset, demonstrating plasticity of their maturation state. This detailed characterization explained the heterogeneity of the chBMDC culture by the simultaneous presence of immature and semi-mature chBMDC subsets, in addition to cells without features of antigen-presenting cells. Our findings are instrumental for the interpretation of experiments using the chBMDC culture in past and future research by providing insights into its phenotypically and functionally distinct cell types.

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Section: Introductionmentioning

confidence: 99%

In vitro Chicken Bone Marrow-Derived Dendritic Cells Comprise Subsets at Different States of Maturation

et al. 2020

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“…In this study, we assessed the ability of DCs pre-exposed to a wellknown reference sensitizer, AHCP, 19,21,23 to activate TCs. For this purpose, DCs were co-cultured with TCs, and TC activation was evaluated through the expression of membrane markers on the TCs, cell proliferation, and cytokine secretion.…”

Section: Discussionmentioning

confidence: 99%

“…18 A DC activation model based on bone marrow-derived DCs (BMDCs) was previously developed in our laboratory, and showed encouraging results for a wide range of reference chemicals. 19,20 In the present study, we aimed to assess the functionality of activated murine DCs by evaluating their ability to prime naive TCs and induce a proliferative response in vitro.…”

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confidence: 99%

Activation of T cells by dendritic cells exposed to a reference sensitizer: Towards a promising model to assess the allergenic potential of chemicals

et al. 2018

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“…The protocol was adapted from an existing protocol . Cells were extracted from the femoral and tibial bone marrow of the mice, and red blood cells were lysed using RBC lysis buffer (eBioscience, Thermo Fisher Scientific).…”

Section: Methodsmentioning

confidence: 99%

“…The protocol was adapted from an existing protocol. 32 Cells were extracted from the femoral and tibial bone marrow of the mice, and red blood cells were lysed using RBC lysis buffer (eBioscience, Thermo Fisher Scientific). Nucleated cells were cultured at 37 °C with 5% CO 2 in BMDC medium: RPMI 1640 medium-GlutaMAX with 1 mM sodium pyruvate, 10 mM HEPES, 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco, Thermo Fisher Scientific), 10% heat-inactivated fetal bovine serum (FBS) (Eurobio Scientific, Les Ulis, France), and 100 μM 2-mercaptoethanol supplemented with 10−50 ng/mL granulocyte-macrophage colony-stimulating factor (GM-CSF) (Miltenyi, Bergisch Gladbach, Germany).…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

Deamidation and Enzymatic Hydrolysis of Gliadins Alter Their Processing by Dendritic Cells in Vitro

Villemin

Tranquet

Solé-Jamault

et al. 2019

J. Agric. Food Chem.

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Gliadins are major wheat allergens. Their treatment by acid or enzymatic hydrolysis has been shown to modify their allergenic potential. As the interaction of food proteins with dendritic cells (DCs) is a key event in allergic sensitization, we wished to investigate whether deamidation and enzymatic hydrolysis influence gliadin processing by DC and to examine the capacity of gliadins to activate DCs. We compared the uptake and degradation of native and modified gliadins by DCs using mouse bone marrow-derived DCs. We also analyzed the effects of these interactions on the phenotypes of DCs and T helper (Th) lymphocytes. Modifying gliadins induced a change in physicochemical properties (molecular weight, hydrophobicity, and sequence) and also in the peptide size. These alterations in turn led to increased uptake and intracellular degradation of the proteins by DCs. Native gliadins (NGs) (100 μg/mL), but not modified gliadins, increased the frequency of DC expressing CD80 (15.41 ± 2.36% vs 6.81 ± 1.10%, p < 0.001), CCR7 (28.53 ± 8.17% vs 17.88 ± 2.53%, p < 0.001), CXCR4 (70.14 ± 4.63% vs 42.82 ± 1.96%, p < 0.001), and CCR7-dependent migration (2.46 ± 1.45 vs 1.00 ± 0.22, p < 0.01) compared with NGs. This was accompanied by Th lymphocyte activation (30.37 ± 3.87% vs 21.53 ± 3.14%, p < 0.1) and proliferation (16.39 ± 3.97% vs 9.31 ± 2.80%, p > 0.1). Moreover, hydrolysis decreases the peptide size and induces an increase in gliadin uptake and degradation. Deamidation and extensive enzymatic hydrolysis of gliadins modify their interaction with DCs, leading to alteration of their immunostimulatory capacity. These findings demonstrate the strong relationship between the biochemical characteristics of proteins and immune cell interactions.

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In vitro detection of chemical allergens: an optimized assay using mouse bone marrow‐derived dendritic cells

Cited by 12 publications

References 59 publications

In vitro Chicken Bone Marrow-Derived Dendritic Cells Comprise Subsets at Different States of Maturation

In vitro Chicken Bone Marrow-Derived Dendritic Cells Comprise Subsets at Different States of Maturation

Activation of T cells by dendritic cells exposed to a reference sensitizer: Towards a promising model to assess the allergenic potential of chemicals

Deamidation and Enzymatic Hydrolysis of Gliadins Alter Their Processing by Dendritic Cells in Vitro

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