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首页> 外文期刊>Frontiers in Microbiology >Protective Efficacy of Coccidial Common Antigen Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) against Challenge with Three Eimeria Species
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Protective Efficacy of Coccidial Common Antigen Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) against Challenge with Three Eimeria Species

机译：球虫常见抗原甘油醛3-磷酸脱氢酶（GAPDH）对三种 Eimeria 物种的攻击防护作用

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Coccidiosis is an intestinal disorder of poultry and often caused by simultaneous infections of several Eimeria species. GAPDH is one of the immunogenic common antigens among Eimeria tenella , E. acervulina , and E. maxima identified in our previous study. The present study was performed to further evaluate its immunogenicity and protective efficacy. The genes of GAPDH cloned from E. acervulina and E. maxima were named as EaGAPDH and EmGAPDH , respectively. The immunogenicity of recombinant proteins of EaGAPDH and EmGAPDH were analyzed by Western blot. The transcription and expression of pVAX-EaGAPDH and pVAX-EmGAPDH in the injected muscles were detected by reverse transcription PCR (RT-PCR) and Western blot, respectively. GAPDH-induced changes of T lymphocytes subpopulation, cytokines production, and antibody were determined using flow cytometry, quantitative real-time PCR (qPCR), and ELISA, respectively. Finally, the protective efficacies of pVAX-EaGAPDH and pVAX-EmGAPDH were evaluated by vaccination and challenge experiments. The results revealed that the recombinant GAPDH proteins reacted with the corresponding chicken antisera. The EaGAPDH genes were successfully transcribed and expressed in the injected muscles. Vaccination with pVAX-EaGAPDH and pVAX-EmGAPDH significantly increased the proportion of CD4~(+)and CD8~(+)T lymphocytes, the cytokines productions of IFN-γ, IL-2, IL-4 et al., and IgG antibody levels compared to controls. The vaccination increased the weight gains, decreased the oocyst outputs, alleviate the enteric lesions compared to controls, and induced moderate anti-coccidial index (ACI). In conclusion, the coccidial common antigen of GAPDH induced significant humoral and cellular immune response and effective protection against E. tenella , E. acervulina , E. maxima , and mixed infection of the three Eimeria species.
机译：球虫病是一种家禽肠道疾病，通常是由几种艾美球虫同时感染引起的。 GAPDH是在我们先前的研究中鉴定的艾美尔球虫，小肠埃希菌和大肠埃希菌中的一种免疫原性常见抗原。进行本研究以进一步评估其免疫原性和保护功效。从小球藻和大肠埃希菌克隆的GAPDH基因分别命名为EaGAPDH和EmGAPDH。通过Western blot分析EaGAPDH和EmGAPDH重组蛋白的免疫原性。分别通过逆转录PCR（RT-PCR）和Western blot检测pVAX-EaGAPDH和pVAX-EmGAPDH在所注射的肌肉中的转录和表达。 GAPDH诱导的T淋巴细胞亚群，细胞因子生成和抗体的变化分别使用流式细胞仪，定量实时PCR（qPCR）和ELISA进行测定。最后，通过疫苗接种和攻击实验评估了pVAX-EaGAPDH和pVAX-EmGAPDH的保护效果。结果显示重组GAPDH蛋白与相应的鸡抗血清反应。 EaGAPDH基因已成功转录并在注射的肌肉中表达。用pVAX-EaGAPDH和pVAX-EmGAPDH进行疫苗接种可显着增加CD4〜（+）和CD8〜（+）T淋巴细胞的比例，IFN-γ，IL-2，IL-4等的细胞因子产生以及IgG抗体水平与对照相比。与对照组相比，接种疫苗可增加体重，减少卵囊产量，减轻肠道损害，并诱导中等的抗球虫指数（ACI）。总之，GAPDH的球菌通用抗原诱导了显着的体液和细胞免疫应答，并有效抵抗了艾美球虫，小球藻，大肠埃希菌和三种艾美球虫的混合感染。

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