Functional characterization of the turkey macrophage migration inhibitory factor

Area of Science:

• Immunology
• Molecular Biology
• Comparative Genomics

Background:

• Macrophage migration inhibitory factor (MIF) is crucial for immune regulation in innate and adaptive immunity.
• Understanding avian MIF functions aids in comparative immunology and disease resistance research.

Purpose of the Study:

• To clone and express the turkey MIF (TkMIF) gene.
• To characterize the basic biological functions of recombinant TkMIF (rTkMIF).
• To investigate potential cross-reactivity between turkey and chicken MIF.

Main Methods:

• TkMIF gene amplification from turkey spleen RNA.
• Cloning into a prokaryotic expression vector (pET11a) and expression in E. coli.
• Purification of rTkMIF via HPLC, followed by SDS-PAGE and Western blot analysis.
• Functional assays including cell migration inhibition and qRT-PCR for cytokine expression.

Main Results:

• The TkMIF gene encodes a 115-amino acid protein with high sequence identity to other avian MIFs (100% with chicken).
• Recombinant TkMIF (rTkMIF) was successfully expressed, purified, and confirmed by Western blot using anti-chicken MIF antisera.
• rTkMIF dose-dependently inhibited mononuclear cell and splenocyte migration and modulated pro-inflammatory cytokine (IFN-γ, IL-17F) and anti-inflammatory cytokine (IL-10, IL-13) expression.

Conclusions:

• Turkey MIF shares high sequence similarity and comparable biological functions with chicken MIF.
• rTkMIF exhibits migration inhibitory activity and influences cytokine profiles, suggesting biological cross-reactivity between turkey and chicken MIF.