What question did this study set out to answer?

The research aims to uncover the mechanisms by which EtMIC2 facilitates E. tenella invasion and inhibits apoptosis of host cells.

April 12, 2026Open Access

PATHOGENIC MECHANISM OF EIMERIA TENELLA EtMIC2 promotes Eimeria tenella invasion and inhibits host cell apoptosis through binding to the ITGAV receptor

Key Points

The research aims to uncover the mechanisms by which EtMIC2 facilitates E. tenella invasion and inhibits apoptosis of host cells.
Utilized His pull-down mass spectrometry to identify interacting proteins.
Conducted KEGG enrichment analysis to reveal apoptosis-related proteins.
Performed co-immunoprecipitation in chicken enterocyte cells to detect EtMIC2 and ITGAV interactions.
Employing Yeast Two-Hybrid assays to confirm protein interactions.
Performed bimolecular fluorescence complementation to visualize interactions in live cells.
EtMIC2 was found to significantly increase E. tenella infection rates and activity of apoptosis-related proteins ITGAV, FAK, and Bcl2.
The intervention of EtMIC2 decreased host cell apoptosis markers such as Caspase 3 activity and increased proliferation pathways.
Knockdown of ITGAV led to a reversal of effects on E. tenella invasion and apoptosis, confirming its role as a mediator.

Abstract

Chicken coccidiosis inflicts severe economic losses on the poultry industry. The mechanisms underlying Eimeria tenella ( E. tenella ) invasion, colonization, and damage to host cells remain incompletely elucidated. This study was designed to elucidate Et MIC2-interacting proteins and the mechanism that EtMIC2 regulates E. tenella invasion and host cell apoptosis. His pull-down-MS and KEGG enrichment results showed apoptosis-related proteins including ITGAV (Integrin alpha-V), PlexinB2, MAP2K6, GNA11, TRK, CASP7, PAK1, and RAP1b. Only the interaction model between EtMIC2 and ITGAV met the AlphaFold2 thresholds (pLDDT > 70, pTM+ipTM > 0.75). The CO-IP results showed that in the primary host cells (CECs) of E. tenella , protein bands corresponding to Et MIC2 (50 kDa) and ITGAV (135 kDa) were successfully detected in the IP group. Blue colony growth on SD/-Ade,-His,-Leu,-Trp/X-α-Gal plates for the group pGBKT7-EtMIC2 or pGBKT7- Et MIC2-3 (153-1029 bp) or pGBKT7- Et MIC2-5 (855-1029 bp) co-transformed with pGADT7-ITGAV in Yeast Two-Hybrid assay. Green fluorescence was observed in DF-1 cells pBiFC-EtMIC2-VN173 or pBiFC-EtMIC2-3-VN173 or pBiFC-EtMIC2-5-VN173 were co-transfected with pBiFC-ITGAV-VC155 in Bimolecular fluorescence complementation. The addition of EtMIC2 protein significantly increased (P < 0.05) E. tenella infection rate, gene expression and protein activity of ITGAV , FAK , PLC , PKC , p65 , PI3K , Akt , ERK , and Bcl2, and reduced host cell Caspase 3 activity, apoptosis rate, gene expression and protein activity of JNK , p38 , Bax , and Caspase 3. Knockdown of ITGAV produced the opposite effects. The function of Et MIC2 was prevented following the knockdown of ITGAV . The above finding indicate that the EtMIC2 855-1029 bp (285-342 aa) region interacts with ITGAV, leading to the upregulation of ITGAV, FAK, PLC, PKC, p65, PI3K, Akt, ERK , and Bcl2 , while downregulating JNK, p38, Bax , and Caspase 3 , thereby promoting E. tenella invasion and inhibiting host cell apoptosis.

PATHOGENIC MECHANISM OF EIMERIA TENELLA EtMIC2 promotes Eimeria tenella invasion and inhibits host cell apoptosis through binding to the ITGAV receptor

Key Points

Abstract

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