Expression of toll-like receptor 4 and its associated cytokines from peripheral blood mononuclear cells in Leghorn chickens

Juthatip Jeenkeawpieam; Chananphat Tantikositruj; Warangkana Kitpipit; Anyarat Thiptara; Autchara Kayan; Kittichai Unjit; Siriluk Sintupachee; Chaiwat Boonkaewwan

doi:10.14202/vetworld.2023.1541-1545

Expression of toll-like receptor 4 and its associated cytokines from peripheral blood mononuclear cells in Leghorn chickens

Vet World. 2023;16(7):1541-1545. doi: 10.14202/vetworld.2023.1541-1545. Epub 2023 Jul 31.

Authors

Juthatip Jeenkeawpieam¹, Chananphat Tantikositruj², Warangkana Kitpipit¹, Anyarat Thiptara³, Autchara Kayan⁴, Kittichai Unjit², Siriluk Sintupachee⁵, Chaiwat Boonkaewwan^{1

6}

Affiliations

¹ Akkhraratchakumari Veterinary College, Walailak University, Nakhon Si Thammarat 80160, Thailand.
² Department of Livestock Development, National Institute of Animal Health, Bangkok 10900, Thailand.
³ Department of Livestock Development, Veterinary Research and Development Center (Upper Southern Region), Nakhon Si Thammarat 80110, Thailand.
⁴ Department of Animal Science, Kasetsart University, Bangkok 10900, Thailand.
⁵ Program in Creative Innovation in Science and Technology, Faculty of Science and Technology, Nakhon Si Thammarat Rajabhat University, Nakhon Si Thammarat, 80280, Thailand.
⁶ One Health Research Center, Walailak University, Nakhon Si Thammarat, 80160, Thailand.

Abstract

Background and aim: Immune cells require toll-like receptor 4 (TLR4) to respond to lipopolysaccharides (LPS) by releasing pro-inflammatory cytokines. Peripheral blood mononuclear cells (PBMCs) are used to assess changes in cytokines released in response to diseases or pathogens. This study aimed to assess TLR4 gene expression in PBMCs from Leghorn chicken and the release of related cytokines.

Materials and methods: Peripheral blood mononuclear cells were isolated from blood samples obtained from Leghorn chicks. The PBMC cultures were stimulated with various concentrations of LPS (0.01-1 μg/ml). Polymerase chain reaction was used to detect TLR4 expression. The production of tumor necrosis factor-alpha (TNF-α) and interleukins (IL-1β and IL-6) was quantified using an enzyme-linked immunosorbent assay.

Results: We found that TLR4 was expressed in both non-stimulated and stimulated Leghorn chicken PBMCs. In addition, the release of TNF-α, IL-1β, and IL-6 levels in Leghorn chicken PBMCs increased significantly with an increase in LPS concentration (0.01-1 μg/mL) (p < 0.05).

Conclusion: Although TLR4 was expressed in both non-stimulated and stimulated Leghorn chicken PBMCs, its expression was significantly higher in LPS-stimulated PBMCs Therefore, the chicken's endotoxin response can be assessed by evaluating the pro-inflammatory cytokine production from PBMCs.

Keywords: Leghorn chicken; Toll-like receptor 4; peripheral blood mononuclear cell; pro-inflammatory cytokine.