细粒棘球绦虫EgM123表面展示型酿酒酵母菌株的构建及免疫原性初步评价

doi:10.11843/j.issn.0366-6964.2025.01.035

Abstract

Abstract:

The objective of this study is to construct a surface-displayed Saccharomyces cerevisiae EBY100/PYD1-EgM123 for Echinococcus granulosus, and to evaluate its immunogenicity. The EgM123 gene of E. granulosus was cloned, and a surface display yeast strain EBY100/PYD1-EgM123 was constructed. The S. cerevisiae strain EBY100/PYD1-EgM123 was examined using immunofluorescence assay, and BCA protein quantitative analysis to determine its localization and quantity. Additionally, experimental mice were immunized with the EBY100/pYD1-EgM123. The immune efficacy of the EBY100/pYD1-EgM123 was evaluated by measuring antibody and cytokine in the serum of the mice. The results showed that the ORF of EgM123 was successfully cloned and its length was 594 bp. The results of PCR and restriction endonuclease analysis showed that the surface-displayed S. cerevisiae EBY100/PYD1-EgM123 was successfully constructed. The results of immunofluorescence showed that the EBY100/PYD1-EgM123 protein exhibited green fluorescence on the surface of the S. cerevisiae. The expression level of EBY100/pYD1-EgM123 was quantified at 50 OD_{600 nm} using the BCA protein quantitative assay. At 72 h, the expression level was found to be 15.59 μg and 0.31 μg/OD_{600 nm}, respectively. The levels of specific IgG, IgA, and IgM in the EBY100/pYD1-EgM123 group were found to be significantly higher than those in the PBS Group, EBY100 Group, and EBY100/PYD1 group (P < 0.05). The results also indicated that the EBY100/PYD1-EgM123 effectively enhances the secretion of IL-17 and IFN-γ (P < 0.001). In conclusion, the present study successfully constructed the surface-displayed S. cerevisiae EBY100/PYD1-EgM123, and demonstrating its ability to stimulate specific immune responses in mice. It is further indicated that it has the potential to be a candidate vaccine against E. granulosus in the final host.

Key words: Echinococcus granulosus, EgM123, surface-displayed Saccharomyces cerevisiae, oral live vector vaccine

CLC Number:

S852.734

JIA Xinyue, MA Jing, PU Na, ZHAO Wenqing, CHEN Xuke, ZHANG Yanyan, SUN Yan, BO Xinwen, WANG Zhengrong. Construction and Evaluation of a Surface-displayed Saccharomyces cerevisiae EBY100/PYD1-EgM123 for Echinococcus granulosus[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(1): 378-391.

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References 28

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方案 Regimen	首次免疫 First immunity	加强免疫 Boost immunity
1	第1、2、3天每天灌服50 μL	第16、17、18天每天灌服50 μL
2	第1、2天每天灌服75 μL	第16、17天每天灌服75 μL
3	第1天灌服150 μL	第16天灌服150 μL

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Construction and Evaluation of a Surface-displayed Saccharomyces cerevisiae EBY100/PYD1-EgM123 for Echinococcus granulosus

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