Functional and Key Gene Expression Analyses of Chicken Monocyte-derived Dendritic Cells with Recombinant Interleukin...

Area of Science:

• Immunology
• Molecular Biology
• Veterinary Science

Background:

• Dendritic cells (DCs) are crucial for bridging innate and adaptive immunity.
• DCs are increasingly utilized in poultry vaccine development.
• Monocyte transformation induced by interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF) is an effective method for DC production.

Purpose of the Study:

• To clone, characterize, and express chicken IL-4 (cIL-4) cDNA in Escherichia coli.
• To utilize the expressed cIL-4 to induce chicken monocytes into monocyte-derived DCs (MDDCs).
• To evaluate the efficiency and timeline of MDDC maturation using bacterially expressed cIL-4.

Main Methods:

• Cloning of the full-length chicken IL-4 (cIL-4) cDNA.
• Expression of cIL-4 in a bacterial host (Escherichia coli).
• Induction of chicken monocytes using expressed cIL-4 and GM-CSF, followed by morphological and gene expression analysis.

Main Results:

• Successful cloning and expression of functional cIL-4 in E. coli.
• Induction of chicken monocytes resulted in cells with typical DC features, including membrane protrusions, by day four.
• Gene expression analysis confirmed a significant shift from monocytes to DCs.
• Mature MDDCs were produced within six to seven days post-cytokine induction.

Conclusions:

• Bacterial expression of cIL-4 is a viable strategy for producing chicken dendritic cells.
• This method provides a potentially efficient and timely approach for generating MDDCs for poultry vaccine applications.
• No codon optimization or further modifications were needed for bacterial cIL-4 expression to yield mature MDDCs.