中国真菌学杂志2025，Vol.20Issue(2)：166-175,10.
白假丝酵母菌抗原在外阴阴道假丝酵母菌病中免疫原性的初步研究
Preliminary study on the immunogenicity of Candida albicans antigen in vulvovaginal candidiasis
摘要
Abstract
Objective Screen Candida albicans antigen by bioinformatics analysis and study its immunogenicity in vulvo-vaginal candidiasis(VVC).Methods ①NCBI,CGD,AlphaFold2,VaxiJen,Expasy,IEDB and other network databases were used to search and analyze the biological information such as gene sequence,protein sequence,protein structure,pro-tein antigenicity,protein physicochemical properties and protein immune epitopes of Candida albicans proteins.② Accord-ing to the predicted results,the target antigen was screened and the recombinant expression vector was constructed using Escherichia coli as the host.The expression of the target protein was induced by the E.coli expression system,and the target protein was purified by His tag method.③Serum samples and vaginal secretion samples of patients with clinically di-agnosed VVC,RVVC and normal people were collected.ELISA was used to detect the levels of antigen-specific antibody IgG in serum samples and antigen-specific antibody sIgA in vaginal secretion samples.Results ① Fifteen candidate antigens of C.albicans were preliminarily screened by literature review and bioinformatics analysis.②Proteins with high immunoge-nicity,strong hydropathy,stable structure,and covering as many immunodominant epitopes as possible were predicted and screened by bioinformatics database.Finally,three single proteins with His tag Eno1,Sap6p,and Ywp1p were constructed.Among them,Eno1 and Sap6p were expressed in soluble form and inclusion body in E.coli BL21(DE3),respectively,and Ywp1p was expressed in soluble form in Rosetta(DE3).After purification by His tag,the target proteins with high purity were obtained.③ELISA was used to detect antigen-specific IgG in serum samples.There was a significant difference in Eno1 protein between VVC group and normal group(P<0.01),Sap6p protein in VVC group and normal group(P<0.001),Ywp1p protein in VVC group and normal group had no statistical difference.There was no significant difference in Eno1,Sap6p,Ywp1p protein between RVVC group and normal group,VVC group and RVVC group.sIgA was not detec-ted in Eno1,Sap6p and Ywp1p protein.Conclusion The recombinant proteins Eno1,Sap6p and Ywp1p containing His tag were successfully constructed,expressed and purified.ELISA results showed that the recombinant proteins Eno1 and Sap6p could be used as candidate antigens for C.albicans vaccine.关键词
白假丝酵母菌/外阴阴道假丝酵母菌病/重组蛋白/免疫原性Key words
Candida albicans/vulvovaginal candidiasis/recombinant proteins/immunogenicity分类
医药卫生引用本文复制引用
杨楠,马守叶,祝秉东,刘会玲..白假丝酵母菌抗原在外阴阴道假丝酵母菌病中免疫原性的初步研究[J].中国真菌学杂志,2025,20(2):166-175,10.基金项目
甘肃省卫生健康行业科研计划项目(GSWSKY2022-17) （GSWSKY2022-17）
甘肃省人民医院院内科研基金项目(21GSSYC-51) （21GSSYC-51）
