Recombinant <i>Culicoides obsoletus</i> complex allergens stimulate antigen‐specific T cells of insect bite hypersensitive Shetland ponies <i>in vitro</i>

Meulenbroeks, Chantal; Meide, Nathalie M.A. van der; Willemse, Ton; Rutten, Victor P.M.G.; Tijhaar, Edwin

doi:10.1111/vde.12251

Cited by 6 publications

(4 citation statements)

References 27 publications

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“…While FBS can in general be considered further for in vitro research, the results here re-emphasize the need for batch testing. These results are very encouraging for the clinical application of equine MoDC and confirm a most recent report using cells generated with horse serum for recall responses [44]. However, the effect of autologous serum or different serum conditions on the phenotype and function of equine MoDC had not been systematically investigated previously.…”

Section: Resultssupporting

confidence: 83%

Equine dendritic cells generated with horse serum have enhanced functionality in comparison to dendritic cells generated with fetal bovine serum

et al. 2016

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BackgroundDendritic cells are professional antigen-presenting cells that play an essential role in the initiation and modulation of T cell responses. They have been studied widely for their potential clinical applications, but for clinical use to be successful, alternatives to xenogeneic substances like fetal bovine serum (FBS) in cell culture need to be found. Protocols for the generation of dendritic cells ex vivo from monocytes are well established for several species, including horses. Currently, the gold standard protocol for generating dendritic cells from monocytes across various species relies upon a combination of GM-CSF and IL-4 added to cell culture medium which is supplemented with FBS. The aim of this study was to substitute FBS with heterologous horse serum. For this purpose, equine monocyte-derived dendritic cells (eqMoDC) were generated in the presence of horse serum or FBS and analysed for the effect on morphology, phenotype and immunological properties. Changes in the expression of phenotypic markers (CD14, CD86, CD206) were assessed during dendritic cell maturation by flow cytometry. To obtain a more complete picture of the eqMoDC differentiation and assess possible differences between FBS- and horse serum-driven cultures, a transcriptomic microarray analysis was performed. Lastly, immature eqMoDC were primed with a primary antigen (ovalbumin) or a recall antigen (tetanus toxoid) and, after maturation, were co-cultured with freshly isolated autologous CD5+ T lymphocytes to assess their T cell stimulatory capacity.ResultsThe microarray analysis demonstrated that eqMoDC generated with horse serum were indistinguishable from those generated with FBS. However, eqMoDC incubated with horse serum-supplemented medium exhibited a more characteristic dendritic cell morphology during differentiation from monocytes. A significant increase in cell viability was also observed in eqMoDC cultured with horse serum. Furthermore, eqMoDC generated in the presence of horse serum were found to be superior in their functional T lymphocyte priming capacity and to elicit significantly less non-specific proliferation.ConclusionsEqMoDC generated with horse serum-supplemented medium showed improved morphological characteristics, higher cell viability and exhibited a more robust performance in the functional T cell assays. Therefore, horse serum was found to be superior to FBS for generating equine monocyte-derived dendritic cells.

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Section: Resultssupporting

confidence: 83%

Equine dendritic cells generated with horse serum have enhanced functionality in comparison to dendritic cells generated with fetal bovine serum

et al. 2016

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“…This might be due to a higher Our study confirmed that a combination of r-allergens differentiates better between IBH-affected and nonaffected horses than the use of single r-allergens or WBE. 16,52,53 Only 45% and 42% of the IBH-affected horses were positive on CO-WBE and CN-TH, respectively, when a cut-off giving a high specificity was used.…”

Section: Discussionmentioning

confidence: 99%

Component‐resolved microarray analysis of IgE sensitization profiles to Culicoides recombinant allergens in horses with insect bite hypersensitivity

Novotny

White

Wilson

et al. 2020

Allergy

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Background Allergy to bites of blood‐sucking insects, including biting midges, can affect both human and veterinary patients. Horses are often suffering from an IgE‐mediated allergic dermatitis caused by bites of midges (Culicoides spp). With the aim to improve allergen immunotherapy (AIT), numerous Culicoides allergens have been produced as recombinant (r‐) proteins. This study aimed to test a comprehensive panel of differently expressed Culicoides r‐allergens on a cohort of IBH‐affected and control horses using an allergen microarray. Methods IgE levels to 27 Culicoides r‐allergens, including 8 previously unpublished allergens, of which 11 were expressed in more than one expression system, were determined in sera from 347 horses. ROC analyses were carried out, cut‐offs selected using a specificity of 95% and seropositivity rates compared between horses affected with insect bite hypersensitivity (IBH) and control horses. The combination of r‐allergens giving the best performing test was determined using logistic regression analysis. Results Seropositivity was significantly higher in IBH horses compared with controls for 25 r‐allergens. Nine Culicoides r‐allergens were major allergens for IBH with seven of them binding IgE in sera from > 70% of the IBH‐affected horses. Combination of these top seven r‐allergens could diagnose > 90% of IBH‐affected horses with a specificity of > 95%. Correlation between differently expressed r‐allergens was usually high (mean = 0.69, range: 0.28‐0.91). Conclusion This microarray will be a powerful tool for the development of component‐resolved, patient‐tailored AIT for IBH and could be useful for the study of allergy to biting midges in humans and other species.

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“…Advances in the identification of the allergens involved in IBH provide the basis for both improved diagnostic methods with specific allergens and for new promising tools for targeted allergen immunotherapy (AIT) using the major allergens of Culicoides spp. [4,11,12,[22][23][24].…”

Section: Introductionmentioning

confidence: 99%

Comparison of Skin Prick Tests (SPT), Intradermal Tests (IDT) and In Vitro Tests in the Characterization of Insect Bite Hypersensitivity (IBH) in a Population of Lusitano Horses: Contribution for Future Implementation of SPT in IBH Diagnosis

Pessoa,

Branco-Ferreira,

Jónsdóttir

et al. 2023

Animals

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Thirty controls (C) and 30 IBH-affected (T) Lusitano horses were evaluated. T horses were included based on anamnesis and physical examination, supported by questionnaires. All horses were submitted to skin tests, Intrademal (IDT) and Skin Prick Tests (SPT), on the neck with 14 specific allergens, 13 recombinant proteins (r-proteins) from Culicoides nubeculosus (Cul n) and Culicoides obsoletus (Cul o) salivary glands and Culicoides nubeculosus Whole Body Extract (Cul n WBE). Addicionally, a cluster of six T and six C horses were also tested with Cul n 3 and Cul n 4 produced in insect cells and barley, as well as E. coli produced Cul o 3 and Cul o WBE. Allergen concentrations were 10 µg/mL for IDT and 100 µg/mL for SPT, and wheal diameters assessed at 20 min, 6 and 48 h. IDTs were considered positive when wheal diameter was ≥50% of the histamine wheal and SPT’s ≥ 0.9 cm. In vitro tests, allergen-specific serum IgE and sulfidoleukotriene (sLT) release assay were also carried out. Results showed that Cul n WBE, Cul n 7, 8, 9, Cul o1P and Cul o 2P were the best performing allergens for SPTs (p ≤ 0.0001) for the 1st allergen panel and Cul o WBE, Cul n 3 Bar and Cul n 4 Bac (p ≤ 0.05) for the 2nd, presenting a higher discriminatory diagnostic potential than IDTs, at a concentration of 100 µg/mL, with readings assessed at 20 min. Regarding in vitro tests overall, the sLT release assay performed best.

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Recombinant Culicoides obsoletus complex allergens stimulate antigen‐specific T cells of insect bite hypersensitive Shetland ponies in vitro

Abstract: The recombinant C. obsoletus complex allergens can stimulate antigen-specific Th1 and IL10 producing Treg cells and are therefore promising candidates for the immunotherapy of IBH.

Cited by 6 publications

References 27 publications

Equine dendritic cells generated with horse serum have enhanced functionality in comparison to dendritic cells generated with fetal bovine serum

Equine dendritic cells generated with horse serum have enhanced functionality in comparison to dendritic cells generated with fetal bovine serum

Component‐resolved microarray analysis of IgE sensitization profiles to Culicoides recombinant allergens in horses with insect bite hypersensitivity

Comparison of Skin Prick Tests (SPT), Intradermal Tests (IDT) and In Vitro Tests in the Characterization of Insect Bite Hypersensitivity (IBH) in a Population of Lusitano Horses: Contribution for Future Implementation of SPT in IBH Diagnosis

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