Analysis of the immunomodulatory properties of mycobacterium cell wall fraction on the cytokine production of peripheral blood mononuclear cells of healthy dogs.

IF 1.9 3区 农林科学 Q3 DERMATOLOGY
Veterinary dermatology Pub Date : 2024-12-01 Epub Date: 2024-08-14 DOI:10.1111/vde.13288
Robert Ward, Geoffrey A Wood, Charlotte Pye, Khalil Karimi, Anthony Yu, Myles St-Denis, Katherine Blake, Sugandha Raj, Samson Oladokun, Shayan Sharif
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Abstract

Background: Mycobacterium cell wall fraction (MCWF) is derived from nonpathogenic Mycobacterium phlei and is used as an immunomodulatory compound in clinical practice, yet its mode-of-action requires further research.

Objective: To evaluate the host response to MCWF in canine peripheral blood mononuclear cells (PBMCs) by using enzyme-linked immunosorbent assays (ELISA) and quantitative reverse transcription (qRT)-PCR for assessment of cytokines.

Animals: Eight healthy Labrador retrievers.

Materials and methods: PBMCs were isolated from whole blood using density centrifugation. The cells were cultured with different concentrations of MCWF or a potent stimulator of cytokine production, phorbol 12-myristate 13-acetate/ionomycin, or left in cell culture medium for 24, 48 and 72 h. Cytokines were measured by ELISA for interleukin (IL)-4, IL-10 and interferon-gamma (IFN-γ), and by qRT-PCR for IL-4, IL-10, IL-13, IFN-γ, tumour necrosis factor alpha (TNF-α) and transforming growth factor-beta.

Results: A significant increase of IL-10 messenger ribonucleic acid (mRNA) was detected at all time points for all concentrations of MCWF (p < 0.05). Protein analysis reflected this finding, with a maximum IL-10 concentration of 300.6 ± 38.3 μg/mL. Compared to the negative control, post-stimulation elevation of IFN-γ mRNA was noted at 24 h with all concentrations of MCWF (p < 0.01), and TNF-α mRNA was increased for 0.5 μg/dL MCWF only at 72 h (p < 0.05).

Conclusions and clinical relevance: MCWF stimulation of PBMCs results in the elevation of both proinflammatory and regulatory cytokine mRNA. Further research into the role of MCWF as a systemically administered regulatory immunomodulator or adjuvant to allergen-specific immunotherapy should be considered.

分析分枝杆菌细胞壁成分对健康狗外周血单核细胞细胞因子产生的免疫调节作用。
背景:分枝杆菌细胞壁组分(MCWF)来自非致病分枝杆菌phlei,在临床实践中被用作免疫调节化合物,但其作用模式还需要进一步研究:通过酶联免疫吸附试验(ELISA)和用于评估细胞因子的定量反转录(qRT)-PCR,评估犬外周血单核细胞(PBMC)中宿主对 MCWF 的反应:八只健康的拉布拉多猎犬:使用密度离心法从全血中分离出 PBMC。用不同浓度的 MCWF 或细胞因子产生的强效刺激物--光稳定剂 12-肉豆蔻酸 13-乙酸酯/洋霉素培养细胞,或将细胞留在细胞培养基中培养 24、48 和 72 小时。细胞因子用 ELISA 法检测白细胞介素(IL)-4、IL-10 和干扰素-γ(IFN-γ),用 qRT-PCR 法检测 IL-4、IL-10、IL-13、IFN-γ、肿瘤坏死因子α(TNF-α)和转化生长因子-β:结果:IL-10 信使核糖核酸(mRNA)在所有浓度的 MCWF 的所有时间点均有明显增加(p 结论和临床意义:MCWF 刺激 PBMCs 会导致促炎细胞因子和调节细胞因子 mRNA 的升高。应考虑进一步研究 MCWF 作为系统给药的调节性免疫调节剂或过敏原特异性免疫疗法辅助剂的作用。
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来源期刊
Veterinary dermatology
Veterinary dermatology 农林科学-兽医学
CiteScore
3.20
自引率
21.40%
发文量
92
审稿时长
12-24 weeks
期刊介绍: Veterinary Dermatology is a bi-monthly, peer-reviewed, international journal which publishes papers on all aspects of the skin of mammals, birds, reptiles, amphibians and fish. Scientific research papers, clinical case reports and reviews covering the following aspects of dermatology will be considered for publication: -Skin structure (anatomy, histology, ultrastructure) -Skin function (physiology, biochemistry, pharmacology, immunology, genetics) -Skin microbiology and parasitology -Dermatopathology -Pathogenesis, diagnosis and treatment of skin diseases -New disease entities
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