Evaluation of antimicrobial peptides and cytokine production in primary keratinocyte cell culture from healthy and atopic beagles

Abstract

Increased secretion of antimicrobial peptides and cytokines is present in atopic skin. The purpose of this study was to compare the production of b-defensin (cBD)3-like, cathelicidin (cCath) and cytokines in atopic and healthy canine keratinocytes. Seven atopic house dust mites (HDMs) sensitive and five healthy age-matched beagles were used. Keratinocytes were stimulated for 24 h, and the supernatant was collected. A significantly higher production of cBD3-like was present at baseline in atopic compared with healthy keratinocytes, but cBD3-like did not increase after stimulation. IL-17 and lipopolysaccharide increased cBD3-like in healthy compared with atopic keratinocytes. cCath increased in both groups after stimulation. Atopic keratinocytes exposed to HDM produced more IL-8 and keratinocyte-derived chemokine-like than healthy keratinocytes. Exposure to HDM induced an increased IL-8 in atopic keratinocytes and a decreased IFN-c in healthy keratinocytes. These results may suggest an over sensitization of atopic keratinocytes and a possible impairment of the cutaneous defense against micro-organisms.

Adjuvanting Allergen Extracts for Sublingual Immunotherapy: Calcitriol Downregulates CXCL8 Production in Primary Sublingual Epithelial Cells

“…Substances that increase TGF-β secretion by sublingual epithelial cells would therefore be interesting candidates for use as adjuvants during SLIT. While production of IL-6 and IL-10 has been observed in other types of epithelial cells (57)(58)(59)(60)(61)(62)(63), no secretion of these cytokines was detected for canine sublingual epithelial cells by ELISA or RNA sequencing. Furthermore, proallergenic mediators associated with the epithelium, TSLP, IL-25, and IL-33 (4,64) and the proinflammatory cytokines IL-1α and IL-1β were not expressed by canine sublingual epithelial cells.…”

Vitamin D shows in vivo efficacy in a placebo‐controlled, double‐blinded, randomised clinical trial on canine atopic dermatitis

“…Some recent in vitro studies have shown the involvement of AMP in AD in both human beings and dogs 57 58 . In human beings, the synthesis of AMPs is increased under the influence of vitamin D 20 and decreased in vitamin D deficiency 59 .…”

Evaluation of the secretion of antimicrobial peptides and antimicrobial effect of skin wash in atopic and healthy dogs: a preliminary study

“…The AMP levels present in the skin washes of atopic and healthy dogs were measured using either a commercially available (S100A8, S100A9 and S100A12 – Neoscientific; Woburn, MA, USA) or in‐house (cBD3‐like/cBD122 and cCath) previously validated, canine‐specific competitive inhibition ELISAs . To normalize the values of each AMP, the amount of total protein present in the skin wash was also determined using a commercially available kit (Micro BCA™ Protein Assay Kit, ThermoFischer Scientific; Grand Island, NY, USA) following the manufacturer's protocol.…”

“…The most commonly studied AMPs have been identified primarily in epithelial tissues, and include β‐defensins (BDs), cathelicidin (Cath) and S100A peptides . Human and canine keratinocytes have been demonstrated to produce AMPs, among others, BDs, Cath and S100A peptides …”

“…Several studies have been published showing an increase in AMP gene expression in canine atopic versus healthy skin . A positive stimulation of AMPs in both healthy and atopic keratinocytes has been shown after incubation with several immunogens, including bacterial proteins . A modified skin wash technique has been reported to be useful for collecting and quantifying AMPs in canine skin wash .…”