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Peer-reviewed veterinary case report

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

Journal:
Veterinary dermatology
Year:
2024
Authors:
Ward, Robert et al.
Affiliation:
Veterinary Allergy Dermatology and Ear Referral Clinic · Canada
Species:
dog

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&#x2009;h. Cytokines were measured by ELISA for interleukin (IL)-4, IL-10 and interferon-gamma (IFN-&#x3b3;), and by qRT-PCR for IL-4, IL-10, IL-13, IFN-&#x3b3;, tumour necrosis factor alpha (TNF-&#x3b1;) 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&#x2009;<&#x2009;0.05). Protein analysis reflected this finding, with a maximum IL-10 concentration of 300.6&#x2009;&#xb1;&#x2009;38.3&#x2009;&#x3bc;g/mL. Compared to the negative control, post-stimulation elevation of IFN-&#x3b3; mRNA was noted at 24&#x2009;h with all concentrations of MCWF (p&#x2009;<&#x2009;0.01), and TNF-&#x3b1; mRNA was increased for 0.5&#x2009;&#x3bc;g/dL MCWF only at 72&#x2009;h (p&#x2009;<&#x2009;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.

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Original publication: https://pubmed.ncbi.nlm.nih.gov/39140285/