Transcriptional changes in LMH cells induced by Eimeria tenella rhoptry kinase family protein 17

Abstract

Though a number of Eimeria tenella rhoptry kinase family proteins have been identified, little is known about their molecular functions. In the present study, the gene fragment encoding the matured peptide of E. tenella rhoptry kinase family protein 17 (EtROP17) was used to construct a recombinant vector, followed by transfection into leghorn male hepatoma (LMH) cells. Then, the transcriptional changes in the transfected cells were determined by RNA-seq. The expression of EtROP17 in LMH cells was validated by both Western blot and indirect immunofluorescence analysis. Our analysis showed that EtROP17 altered the expression of 309 genes (114 downregulated genes and 195 upregulated genes) in LMH cells. The quantitative real-time polymerase chain reaction (qRT-PCR) results of the selected differentially expressed genes (DEGs) were consistent with the RNA-seq data. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that DEGs were significantly enriched in nine pathways, such as toll-like receptor signaling pathway, ECM-receptor interaction, intestinal immune network for IgA production and focal adhesion. These findings reveal several potential roles of EtROP17, which contribute to understanding the molecular mechanisms underlying the host-parasite interplay.

The Immunoprotective Effect of ROP27 Protein of Eimeria tenella

“…EtROP plays an important role in the parasitic invasion of host cells, the modification of host vacuoles, and the regulation of host cells, and it has been proven to be a key virulence factor. It also has the properties of a protective antigen, and its corresponding antibodies can inhibit the infection of the host by coccidia [32]. Studies have confirmed the expression of EtROP17 in E. tenella merozoites.…”

“…They interact with the host cell through signal transduction, causing a series of downstream effects and becoming a key determinant of protozoan virulence. After entering the cell, ROPs can disrupt host cell signaling and defense mechanisms and assist in recruiting host organelles [32].…”

Bioinformatics-based prediction of biochemical characteristic and potential immunogenic epitopes for ROPs proteins of the E. tenella as candidate vaccine target

“…EtROP plays a crucial role in the invasion process of the mentioned parasite to cells, the alteration of host vacuoles, and the regulatory process of cells, and there are some types of proof for it to be a vital virulence factor. It possesses characteristics of an antigen causing protection, and its corresponding antibodies are capable of the coccidiosis inhibition within the host [53]. However, we have limited knowledge on how the E. tenella ROPs proteins can cause immunoprotection, and only few vaccination research based studies were about these molecules.…”

Transcriptomic analysis of LMH cells in response to the overexpression of a protein of Eimeria tenella encoded by the locus ETH_00028350

“…Total RNA was extracted from E. tenella SD-01 strain, followed by cDNA generation. The LMH cells were grown under the normal culture condition (5% CO 2 , 37°C) in Dulbecco's modified eagle medium (DMEM) containing 10% fetal bovine serum (FBS), 100 μg/mL streptomycin, and 100U/mL penicillin ( 16 ).…”

“…Following construction of transcriptomic libraries, the paired-end sequencing was carried out by Novogene Corporation (Beijing, China). After removing low-quality reads from raw data and reads containing adapter or ploy-N, high-quality clean reads were mapped against the chicken ( Gallus gallus ) reference genome ( 16 ).…”