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IFN-γ inhibitory molecules derived from Eimeria maxima inhibit IL-12 secretion by modulating MAPK pathways in chicken
Chen Chen1, Yufeng Chen1, Mingmin Lu1
1Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China.
Abstract:
IFN-γ plays a crucial role in resisting intracellular parasitic protozoa, such as Eimeria species. In our previous study, we identified 4 molecules derived from Eimeria maxima (E. maxima) that significantly inhibited IFN-γ production. However, the mechanism underlying this inhibitory effect remains unknown. In this study, we first investigated the effects of these 4 IFN-γ inhibitory molecules on the expression levels of chicken Toll-like receptors (chTLRs), IL-12, IL-10, TGF-β, and TNF-α in chicken macrophage HD11 and bone marrow-derived dendritic cells (BMDCs). The results demonstrated that these 4 inhibitory molecules significantly downregulated the mRNA levels of chTLR-2, chTLR-4, chTLR-21, and both mRNA and protein levels of IL-12. Subsequently, to clarify the effects of these 4 inhibitory molecules on the IL-12 secretion-related signaling pathways in chicken macrophages, qRT-PCR and Western blot were used to detect the changes of key molecules involved in the signaling pathways of IL-12 secretion (NF-κB, ERK1/2, p38, JNK, STAT3) following coincubation with these inhibitory molecules. Finally, RNAi was employed to verify the function of key molecules in the signaling pathway. The results revealed a significant upregulation in the expression of ERK1/2 phosphorylated protein induced by the 4 inhibitory molecules. Knockdown of the ERK1/2 gene significantly reduced the inhibitory effect of the 4 E. maxima inhibitory molecules on IL-12. These findings indicate that the 4 inhibitory molecules can inhibit the secretion of IL-12 by upregulating the expression of ERK1/2 phosphorylated protein, which is a key molecule in the ERK-MAPK pathway. Our study may contribute to elucidating the mechanisms underlying immune evasion during E. maxima infections, thereby providing new insights for the control of chicken coccidiosis.
Insights
Four molecules from Eimeria maxima inhibit chicken immune responses by downregulating IL-12 secretion via the ERK1/2 pathway. This discovery offers new strategies for controlling chicken coccidiosis.
Area of Science:
- Immunology
- Parasitology
- Molecular Biology
Background:
- Interferon-gamma (IFN-γ) is vital for combating protozoan parasites like Eimeria species.
- Previous research identified four Eimeria maxima molecules that suppress IFN-γ production, but the mechanism was unclear.
Purpose of the Study:
- To investigate how four Eimeria maxima molecules affect chicken immune gene expression and signaling pathways.
- To elucidate the molecular mechanism by which these molecules inhibit Interleukin-12 (IL-12) secretion.
Main Methods:
- Quantitative reverse transcription PCR (qRT-PCR) and Western blotting were used to analyze gene and protein expression.
- Chicken macrophage (HD11) and bone marrow-derived dendritic cells (BMDCs) were utilized.
- RNA interference (RNAi) was employed to validate key signaling molecules.
Main Results:
- The four molecules downregulated chicken Toll-like receptors (chTLRs) 2, 4, and 21, and suppressed IL-12 mRNA and protein levels.
- These molecules upregulated phosphorylated ERK1/2 protein expression.
- Knockdown of ERK1/2 reversed the inhibitory effect of the molecules on IL-12.
Conclusions:
- The four Eimeria maxima molecules inhibit IL-12 secretion by upregulating ERK1/2 phosphorylation within the ERK-MAPK pathway.
- This study provides insights into Eimeria maxima immune evasion strategies.
- Findings may aid in developing new methods to control chicken coccidiosis.
