Immunomodulatory effects of thymol and cinnamaldehyde in chicken cell lines

Abstract

Thymol and cinnamaldehyde are phytogenic feed additives that have been developed to improve gut health, immunity and growth performance in poultry and swine. This study evaluated the immune modulating effects of a thymol and cinnamaldehyde blend (TCB) in the intestinal system of poultryin vitro, using two chicken cell lines, LMH (liver cell line) which has been used to mimic epithelial cell responses, and HD-11 (monocyte/macrophage-like). Cells with high viability (>95%) from established cell lines were cultured in the presence of TCB at concentrations ranging from 1 ng/ml to 100 ng/ml. The viability, transepithelial electrical resistance (TEER) and phagocytic capacity of co-cultured LMH cells, with or without stimulation with lipopolysaccharide (LPS), was subsequently evaluated. The expression of cytokines, chemokines and pattern recognition receptors by HD-11 monocytes/macrophages was measured by RT-PCR and by proteomic analysis. TCB was well tolerated by both cell lines (cell viability >90% after co-culture with TCB at 100 ng/ml for 48 h with or without LPS). Epithelial integrity of LMH cells (as assessed by TEER) was increased by TCB (10 ng/ml) after 4 h incubation, versus untreated controls, and phagocytic capacity of HD-11 cells was increased, in a dose-dependent manner (P<0.05). In HD-11 cells, TCB (10 ng/ml) downregulated the relative expression of pro-inflammatory cytokines interleukin (IL)-1β, IL-6, IL-8 and the transcription factor cyclooxygenase-2 and upregulated expression of anti-inflammatory IL-10, versus untreated controls (P<0.05). In summary, under the tested conditions, TCB enhanced the epithelial barrier integrity of poultry hepatocytes, increased phagocytic activity and production of anti-inflammatory cytokines by monocytes and macrophages. These results indicated how supplementing TCB in poultry diets can increase bird performance, by increasingin vivo cell membrane integrity (especially important in the gut) and assisting in immune responses, which can liberate energy for growth.