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特定粉尘螨提取物用于犬类免疫治疗。

Specific Dermatophagoides farinae extract for canine immunotherapy.

机构信息

R&D Allergy & Immunology Unit, Laboratorios LETI, S.L, Calle del Sol no. 5, Tres Cantos, Madrid, 28760, Spain.

Animal Health B.U, Laboratorios LETI, S.L, Gran Via de les Corts Catalanes 184, Barcelona, 08038, Spain.

出版信息

Vet Dermatol. 2021 Apr;32(2):131-e29. doi: 10.1111/vde.12902. Epub 2020 Oct 28.

Abstract

BACKGROUND

Canine atopic dermatitis (cAD) is a pruritic allergic skin disease most often caused by Dermatophagoides farinae. Differences in the sensitization profile to D. farinae have been reported between people and dogs. However, allergic dogs traditionally have been treated with extracts intended for human immunotherapy.

HYPOTHESIS/OBJECTIVES: To develop a specific allergen immunotherapy for veterinary practice enriched in canine major allergens and to demonstrate its in vitro efficacy.

ANIMALS

Twenty privately owned dogs, clinically diagnosed with cAD, and three healthy dogs.

METHODS AND MATERIALS

A veterinary D. farinae allergen extract was manufactured and characterized compared to D. farinae extract used for human immunotherapy. The protein profile was analysed by SDS-PAGE and size exclusion chromatography and Der f 15 and Der f 18 allergens quantified by mass spectrometry. The allergenic profile was studied by immunoblot and the biological potency by enzyme-linked immunosorbent assay-inhibition assays. The extract's capacity to induce cytokine production [interleukin (IL)-10, interferon (IFN)-Ɣ] by peripheral blood mononuclear cells also was evaluated.

RESULTS

The veterinary extract showed a higher content of high molecular weight proteins, preferentially recognized by atopic dog sera. The fold-increases in Der f 15 and Der f 18 with respect to the human extract were 2.07 ± 0.32 and 1.63 ± 0.15, respectively. The veterinary extract showed higher biological potency (0.062 versus 0.132 µg required for 50% inhibition of dogs sera) compared to the human extract and induced significantly higher levels of IL-10 (1,780 pg/mL) and IFN-Ɣ (50.4 pg/mL) with respect to the negative control.

CONCLUSIONS AND CLINICAL IMPORTANCE

A veterinary D. farinae extract with a higher content of dog major allergens was developed and in vitro efficacy demonstrated by immunological parameters.

摘要

背景

犬特应性皮炎(cAD)是一种瘙痒性过敏性皮肤病,最常见的病因是粉尘螨。人与犬之间已报道了对粉尘螨的致敏谱存在差异。然而,传统上,过敏的犬类一直接受用于人类免疫疗法的提取物治疗。

假设/目的:为兽医实践开发一种富含犬主要过敏原的特定过敏原免疫疗法,并证明其体外疗效。

动物

20 只私人拥有的、临床诊断为 cAD 的犬和 3 只健康犬。

方法和材料

制造并比较了兽医粉尘螨过敏原提取物与用于人类免疫疗法的粉尘螨提取物。通过 SDS-PAGE 和尺寸排阻色谱法分析蛋白质谱,并通过质谱法定量测定 Der f 15 和 Der f 18 过敏原。通过免疫印迹研究过敏原谱,通过酶联免疫吸附抑制试验评估生物效价。还评估了提取物诱导外周血单核细胞产生细胞因子(白细胞介素(IL)-10、干扰素(IFN)-γ)的能力。

结果

兽医提取物显示出更高含量的高分子量蛋白,这些蛋白更优先被特应性犬血清识别。与人类提取物相比,Der f 15 和 Der f 18 的倍数增加分别为 2.07±0.32 和 1.63±0.15。兽医提取物与人类提取物相比具有更高的生物效价(抑制犬血清所需的 0.062 与 0.132μg),并且与阴性对照相比,诱导的 IL-10(1780pg/mL)和 IFN-γ(50.4pg/mL)水平显著更高。

结论和临床意义

开发了一种含有更高含量犬主要过敏原的兽医粉尘螨提取物,并通过免疫学参数证明了其体外疗效。

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