Characterization of the Antigenic and Immunogenic Properties of the Gametocyte Antigen 56 from <i>Eimeria necatrix</i>

Coccidiosis, caused by <i>Eimeria</i> spp., significantly reduces poultry productivity and causes major economic losses. Traditional control methods are limited by drug resistance and high production costs. Recent genomic and bioinformatic advances have enabled the identification of nove...

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Main Authors: Feiyan Wang, Liqin Cao, Lele Wang, Jinjun Xu, Jianping Tao, Dandan Liu
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Animals
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Online Access:https://www.mdpi.com/2076-2615/15/12/1750
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author Feiyan Wang
Liqin Cao
Lele Wang
Jinjun Xu
Jianping Tao
Dandan Liu
author_facet Feiyan Wang
Liqin Cao
Lele Wang
Jinjun Xu
Jianping Tao
Dandan Liu
author_sort Feiyan Wang
collection DOAJ
description Coccidiosis, caused by <i>Eimeria</i> spp., significantly reduces poultry productivity and causes major economic losses. Traditional control methods are limited by drug resistance and high production costs. Recent genomic and bioinformatic advances have enabled the identification of novel antigens, making recombinant subunit vaccines a promising next-generation strategy by eliciting robust cellular and humoral immune responses. This study investigates the <i>E. necatrix</i> gametocyte protein 56 (EnGAM56) as a potential candidate for recombinant subunit vaccines. The full-length <i>E. necatrix</i> gametocyte <i>gam56</i> gene (En<i>gam56</i>-F) was amplified, expressed in vitro, and characterized via SDS-PAGE and Western blot. Immunofluorescence assays revealed that EnGAM56-F is specifically localized in gametocytes and unsporulated oocysts. Chickens immunized with recombinant proteins (rEnGAM56-F and rEnGAM56-T) were evaluated for immunoprotection against <i>E. necatrix</i> infection through lesion scores, weight gain, oocyst production, anticoccidial index (ACI), and antibody and cytokine levels. The synergistic effects were evaluated by employing various combinations of recombinant proteins, including rEtGAM22, rEtGAM56-T, and rEtGAM59. Results showed that EnGAM56-F encodes a 468-amino acid protein with distinct tyrosine-serine-rich and proline-methionine-rich regions. rEnGAM56-F was specifically recognized by both anti-6 × His tag antibodies and convalescent serum from chickens infected with <i>E. necatrix</i>. Both rEnGAM56-F and rEnGAM56-T provided immune protection, with rEnGAM56-T showing superior efficacy. The combination of rEnGAM (22 + 59 + 56-T) yielded the strongest immune response, followed by rEnGAM (22 + 56-T). These findings highlight the potential of EnGAM56 as a candidate for recombinant subunit anticoccidial vaccines.
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spelling doaj-art-76d60cbd17f54063af2d63b2cf8278c72025-08-20T03:30:29ZengMDPI AGAnimals2076-26152025-06-011512175010.3390/ani15121750Characterization of the Antigenic and Immunogenic Properties of the Gametocyte Antigen 56 from <i>Eimeria necatrix</i>Feiyan Wang0Liqin Cao1Lele Wang2Jinjun Xu3Jianping Tao4Dandan Liu5College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, ChinaJiu Long Animal Husbandry and Veterinary Station, Taizhou 225300, ChinaCollege of Veterinary Medicine, Yangzhou University, Yangzhou 225009, ChinaCollege of Veterinary Medicine, Yangzhou University, Yangzhou 225009, ChinaCollege of Veterinary Medicine, Yangzhou University, Yangzhou 225009, ChinaCollege of Veterinary Medicine, Yangzhou University, Yangzhou 225009, ChinaCoccidiosis, caused by <i>Eimeria</i> spp., significantly reduces poultry productivity and causes major economic losses. Traditional control methods are limited by drug resistance and high production costs. Recent genomic and bioinformatic advances have enabled the identification of novel antigens, making recombinant subunit vaccines a promising next-generation strategy by eliciting robust cellular and humoral immune responses. This study investigates the <i>E. necatrix</i> gametocyte protein 56 (EnGAM56) as a potential candidate for recombinant subunit vaccines. The full-length <i>E. necatrix</i> gametocyte <i>gam56</i> gene (En<i>gam56</i>-F) was amplified, expressed in vitro, and characterized via SDS-PAGE and Western blot. Immunofluorescence assays revealed that EnGAM56-F is specifically localized in gametocytes and unsporulated oocysts. Chickens immunized with recombinant proteins (rEnGAM56-F and rEnGAM56-T) were evaluated for immunoprotection against <i>E. necatrix</i> infection through lesion scores, weight gain, oocyst production, anticoccidial index (ACI), and antibody and cytokine levels. The synergistic effects were evaluated by employing various combinations of recombinant proteins, including rEtGAM22, rEtGAM56-T, and rEtGAM59. Results showed that EnGAM56-F encodes a 468-amino acid protein with distinct tyrosine-serine-rich and proline-methionine-rich regions. rEnGAM56-F was specifically recognized by both anti-6 × His tag antibodies and convalescent serum from chickens infected with <i>E. necatrix</i>. Both rEnGAM56-F and rEnGAM56-T provided immune protection, with rEnGAM56-T showing superior efficacy. The combination of rEnGAM (22 + 59 + 56-T) yielded the strongest immune response, followed by rEnGAM (22 + 56-T). These findings highlight the potential of EnGAM56 as a candidate for recombinant subunit anticoccidial vaccines.https://www.mdpi.com/2076-2615/15/12/1750<i>Eimeria necatrix</i>gametocyte antigensrecombinant subunit vaccineImmunoprotection
spellingShingle Feiyan Wang
Liqin Cao
Lele Wang
Jinjun Xu
Jianping Tao
Dandan Liu
Characterization of the Antigenic and Immunogenic Properties of the Gametocyte Antigen 56 from <i>Eimeria necatrix</i>
Animals
<i>Eimeria necatrix</i>
gametocyte antigens
recombinant subunit vaccine
Immunoprotection
title Characterization of the Antigenic and Immunogenic Properties of the Gametocyte Antigen 56 from <i>Eimeria necatrix</i>
title_full Characterization of the Antigenic and Immunogenic Properties of the Gametocyte Antigen 56 from <i>Eimeria necatrix</i>
title_fullStr Characterization of the Antigenic and Immunogenic Properties of the Gametocyte Antigen 56 from <i>Eimeria necatrix</i>
title_full_unstemmed Characterization of the Antigenic and Immunogenic Properties of the Gametocyte Antigen 56 from <i>Eimeria necatrix</i>
title_short Characterization of the Antigenic and Immunogenic Properties of the Gametocyte Antigen 56 from <i>Eimeria necatrix</i>
title_sort characterization of the antigenic and immunogenic properties of the gametocyte antigen 56 from i eimeria necatrix i
topic <i>Eimeria necatrix</i>
gametocyte antigens
recombinant subunit vaccine
Immunoprotection
url https://www.mdpi.com/2076-2615/15/12/1750
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