Recombinant GMA56 and ROP17 of Eimeria magna conferred protection against infection by homologous species

One of the most common rabbits coccidia species, Eimeria magna is mainly parasitic in the ileal and jejunal epithelial cells. E. magna infection can affect the growth performance of rabbits or cause other secondary diseases. Traditional methods of anticoccidial treatment typically result in drug res...

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Main Authors: Jie Xiao, Hao Chen, Ruoyu Zheng, Jiayan Pu, Xiaobin Gu, Yue Xie, Ran He, Jing Xu, Bo Jing, Xuerong Peng, Guangyou Yang
Format: Article
Language:English
Published: Frontiers Media S.A. 2023-01-01
Series:Frontiers in Immunology
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2022.1037949/full
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author Jie Xiao
Hao Chen
Ruoyu Zheng
Jiayan Pu
Xiaobin Gu
Yue Xie
Ran He
Jing Xu
Bo Jing
Xuerong Peng
Guangyou Yang
author_facet Jie Xiao
Hao Chen
Ruoyu Zheng
Jiayan Pu
Xiaobin Gu
Yue Xie
Ran He
Jing Xu
Bo Jing
Xuerong Peng
Guangyou Yang
author_sort Jie Xiao
collection DOAJ
description One of the most common rabbits coccidia species, Eimeria magna is mainly parasitic in the ileal and jejunal epithelial cells. E. magna infection can affect the growth performance of rabbits or cause other secondary diseases. Traditional methods of anticoccidial treatment typically result in drug resistance and drug residue. Therefore, vaccination is a promising alternative. Gametocyte antigen 56 (GAM56) and rhoptry kinase family proteins (ROPs) are involved in oocyst wall formation and parasite invasion, respectively. A virulence factor, ROP17 contains a serine/threonine kinase catalytic domain. In this study, recombinant E. magna GAM56 (rEmGAM56) and ROP17 (rEmROP17) proteins were obtained from a prokaryotic expression system and their reactogenicity was investigated with immunoblotting. To assess the potential of rEmGAM56 and rEmROP17 as coccidiosis vaccines, New Zealand White rabbits were subcutaneously immunized with 100 μg rEmGAM56 (rGC group) or rEmROP17 (rRC group) twice at 2-week intervals followed by homologous oocyst challenge. The rabbit serum was collected weekly to detect the specific antibody levels. The cytokine levels of pre-challenge serum were measured by enzyme-linked immunosorbent assay and the rabbits were observed and recorded post-challenge for the onset of clinical symptoms. The weight gain, oocyst output, and feed conversion ratio were calculated at the end of the experiment. The results showed that both rEmGAM56 and rEmROP17 had good reactogenicity. The rEmGAM56- or rEmROP17-immunized rabbits had milder clinical symptoms and feed conversion ratios of 3.27:1 and 3.37:1, respectively. The rEmGAM56-immunized rabbits had 81.35% body weight gain and 63.85% oocyst output reduction; the rEmROP17-immunized rabbits had 79.03% body weight gain and 80.10% oocyst output reduction. The ACI of rGC and rRC groups were 162.35 and 171.03, respectively. The specific antibody levels increased rapidly after immunization. Significantly increased interleukin (IL)-2, interferon (IFN)-γ, and IL-17 levels were evident in the rGC and rRC groups (p < 0.05). The rEmGAM56 and rEmROP17 elicited humoral and cellular responses, which protected against E. magna infection in rabbits. Thus, rEmGAM56 and rEmROP17 are potential vaccine candidates against E. magna, and rEmROP17 performed better than rEmGAM56.
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spelling doaj.art-af7d4092e1014a3badb0261df2d388352023-01-12T06:21:16ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-01-011310.3389/fimmu.2022.10379491037949Recombinant GMA56 and ROP17 of Eimeria magna conferred protection against infection by homologous speciesJie Xiao0Hao Chen1Ruoyu Zheng2Jiayan Pu3Xiaobin Gu4Yue Xie5Ran He6Jing Xu7Bo Jing8Xuerong Peng9Guangyou Yang10Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, ChinaDepartment of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, ChinaDepartment of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, ChinaDepartment of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, ChinaDepartment of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, ChinaDepartment of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, ChinaDepartment of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, ChinaDepartment of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, ChinaDepartment of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, ChinaDepartment of Chemistry, College of Life and Basic Science, Sichuan Agricultural University, Chengdu, ChinaDepartment of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, ChinaOne of the most common rabbits coccidia species, Eimeria magna is mainly parasitic in the ileal and jejunal epithelial cells. E. magna infection can affect the growth performance of rabbits or cause other secondary diseases. Traditional methods of anticoccidial treatment typically result in drug resistance and drug residue. Therefore, vaccination is a promising alternative. Gametocyte antigen 56 (GAM56) and rhoptry kinase family proteins (ROPs) are involved in oocyst wall formation and parasite invasion, respectively. A virulence factor, ROP17 contains a serine/threonine kinase catalytic domain. In this study, recombinant E. magna GAM56 (rEmGAM56) and ROP17 (rEmROP17) proteins were obtained from a prokaryotic expression system and their reactogenicity was investigated with immunoblotting. To assess the potential of rEmGAM56 and rEmROP17 as coccidiosis vaccines, New Zealand White rabbits were subcutaneously immunized with 100 μg rEmGAM56 (rGC group) or rEmROP17 (rRC group) twice at 2-week intervals followed by homologous oocyst challenge. The rabbit serum was collected weekly to detect the specific antibody levels. The cytokine levels of pre-challenge serum were measured by enzyme-linked immunosorbent assay and the rabbits were observed and recorded post-challenge for the onset of clinical symptoms. The weight gain, oocyst output, and feed conversion ratio were calculated at the end of the experiment. The results showed that both rEmGAM56 and rEmROP17 had good reactogenicity. The rEmGAM56- or rEmROP17-immunized rabbits had milder clinical symptoms and feed conversion ratios of 3.27:1 and 3.37:1, respectively. The rEmGAM56-immunized rabbits had 81.35% body weight gain and 63.85% oocyst output reduction; the rEmROP17-immunized rabbits had 79.03% body weight gain and 80.10% oocyst output reduction. The ACI of rGC and rRC groups were 162.35 and 171.03, respectively. The specific antibody levels increased rapidly after immunization. Significantly increased interleukin (IL)-2, interferon (IFN)-γ, and IL-17 levels were evident in the rGC and rRC groups (p < 0.05). The rEmGAM56 and rEmROP17 elicited humoral and cellular responses, which protected against E. magna infection in rabbits. Thus, rEmGAM56 and rEmROP17 are potential vaccine candidates against E. magna, and rEmROP17 performed better than rEmGAM56.https://www.frontiersin.org/articles/10.3389/fimmu.2022.1037949/fullEimeria magnagametocyte antigen 56rhoptry kinase family protein 17recombinant proteinsprotective effect
spellingShingle Jie Xiao
Hao Chen
Ruoyu Zheng
Jiayan Pu
Xiaobin Gu
Yue Xie
Ran He
Jing Xu
Bo Jing
Xuerong Peng
Guangyou Yang
Recombinant GMA56 and ROP17 of Eimeria magna conferred protection against infection by homologous species
Frontiers in Immunology
Eimeria magna
gametocyte antigen 56
rhoptry kinase family protein 17
recombinant proteins
protective effect
title Recombinant GMA56 and ROP17 of Eimeria magna conferred protection against infection by homologous species
title_full Recombinant GMA56 and ROP17 of Eimeria magna conferred protection against infection by homologous species
title_fullStr Recombinant GMA56 and ROP17 of Eimeria magna conferred protection against infection by homologous species
title_full_unstemmed Recombinant GMA56 and ROP17 of Eimeria magna conferred protection against infection by homologous species
title_short Recombinant GMA56 and ROP17 of Eimeria magna conferred protection against infection by homologous species
title_sort recombinant gma56 and rop17 of eimeria magna conferred protection against infection by homologous species
topic Eimeria magna
gametocyte antigen 56
rhoptry kinase family protein 17
recombinant proteins
protective effect
url https://www.frontiersin.org/articles/10.3389/fimmu.2022.1037949/full
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