Molecular characterization of a potential receptor of Eimeria acervulina microneme protein 3 from chicken duodenal epithelial cells

Abstract

Eimeria acervulina is one of seven Eimeria spp. that can infect chicken duodenal epithelial cells. Eimeria microneme protein 3 (MIC3) plays a vital role in the invasion of host epithelial tissue by the parasite. In this study, we found that chicken (Gallus gallus) ubiquitin conjugating enzyme E2F (UBE2F) could bind to the MIC3 protein of E. acervulina (EaMIC3), as screened using the yeast two-hybrid system, and that it might be the putative receptor protein of EaMIC3. The UBE2F gene was cloned from chicken duodenal epithelial cells. The recombinant protein of UBE2F (rUBE2F) was expressed in E. coli and the reactogenicity of rUBE2F was analyzed by Western blot. Gene sequencing revealed that the opening reading frame (ORF) of UBE2F was 558 base pairs and encoded a protein of 186 amino acids with a molecular weight of 20.46 kDa. The predicted UBE2F protein did not contain signal peptides or a transmembrane region, but had multiple O-glycosylation and phosphorylation sites. A phylogenetic analysis showed that the chicken UBE2F protein is closely related to those of quail and pigeon (Coturnix japonica and Columba livia). A sporozoite invasion-blocking assay showed that antisera against rUBE2F significantly inhibited the invasion of E. acervulina sporozoites in vitro. Animal experiments indicated that the antisera could significantly enhance average body weight gains and reduce mean lesion scores following a challenge with E. acervulina. These results therefore imply that the chicken UBE2F protein might be the target receptor molecule of EaMIC3 that is involved in E. acervulina invasion.

Identification and characterization of the receptors of a microneme adhesive repeat domain of Eimeria maxima microneme protein 3 in chicken intestine epithelial cells

“…In this study, we assessed the inhibitory efficacy of antibodies targeting EmMAR2 receptors against E. maxima infections in chickens. To achieve this, we employed the method of administration of polyclonal antibodies to conduct the passive immunization trial as previously described ( Crane et al, 1988 ; Wallach et al, 1990 ; Zhang, et al, 2020 ). Notably, in earlier investigations involving passive immunization of chickens against coccidia, the administration of wings intravenously over a period of 5 to 7 consecutive days did not yield significant levels of adverse effects among the immunized chickens ( Wallach et al, 1990 ; Li et al, 2020 ; Zhang et al, 2020 ).…”

“…To achieve this, we employed the method of administration of polyclonal antibodies to conduct the passive immunization trial as previously described ( Crane et al, 1988 ; Wallach et al, 1990 ; Zhang, et al, 2020 ). Notably, in earlier investigations involving passive immunization of chickens against coccidia, the administration of wings intravenously over a period of 5 to 7 consecutive days did not yield significant levels of adverse effects among the immunized chickens ( Wallach et al, 1990 ; Li et al, 2020 ; Zhang et al, 2020 ). Similar to the mechanism underlying passive immunization administered as a therapeutic intervention, the introduction of specific antibodies targeting 4 receptors into the gut of chickens may serve to hinder the interactions between EmMAR2 and these receptors.…”

“…Except for the unchallenged control group, all groups were subjected to oral infection with 1×10 5 sporulated oocysts of E. maxima . Concurrently, intravenous injections were administered into the wings of each chicken once a day for 7 d, as previously described ( Crane et al, 1988 ; Wallach et al, 1990 ; Zhang et al, 2020 ). These injections included 0.5 mL of antibodies targeting rATP6V1G1, rREEP5, rTMED2, rDEGS1, and rpET-32a tag protein, mixed antibodies against all EmMAR2 receptors, and normal rat serum.…”

Eimeria acervulina Microneme Protein 3 Inhibits Apoptosis of the Chicken Duodenal Epithelial Cell by Targeting the Casitas B-Lineage Lymphoma Protein

“…C-Cbl can degrade Fas and DR4/5 by ubiquitination and then inhibit cell apoptosis in T-lymphoma and prostate cancer. Interference of c-Cbl expression can promote apoptosis by upregulating the expression of Fas and DR4/5 ( 33 , 34 ). Also, c-Cbl can degrade caspase-8 by ubiquitination and then inhibit cell apoptosis in gastric cancer ( 35 ).…”

Evaluation of the immune protective effects of rEmMIC2 and rEmMIC3 from Eimeria magna in rabbits

“…BLASTP (Zhang et al 2020a ) was used to study sequence similarity. IEDB (González-Díaz et al 2014 ) was used to predict B-cell antigenic epitopes and conserved domains (Marchler-Bauer et al 2012 ) to predict conserved structural domains.…”