CAJ | 학술논문

目的：预测和鉴定梅毒螺旋体（Tp）膜蛋白TprF氨基端保守区（TprFN ）的优势B细胞表位，为深入研究梅毒多价表位疫苗提供依据。方法从GenBank获取T prFN的氨基酸序列，采用M obyle、ABCpred和IEDB在线软件综合分析预测TprFN的B细胞表位并人工合成多肽；表达重组蛋白 TprFN并经Western blot鉴定后免疫兔，获取血清并测定抗体效价；以TprFN免疫兔血清、梅毒患者血清（设正常人血清和正常兔血清为阴性对照），间接ELISA测定预测的7条人工合成的B细胞表位多肽的免疫反应性和特异性。结果软件综合预测 TprFN的 P1（43‐62AA）、P2（57‐71AA）、P3（81‐88AA）、P4（89‐103AA）、P5（125‐138AA）、P6（231‐251AA）、P7（268‐279AA）可能为B细胞表位；表达一可溶性蛋白，WB鉴定为目的蛋白，其免疫抗体效价为1∶12800以上；ELISA结果显示，预测表位P1、P3与TprFN免疫兔血清及梅毒患者血清均呈阳性反应，而与对照血清均不反应。结论 P1、P3为T prF潜在的优势B细胞表位。
목적：예측화감정매독라선체（Tp）막단백TprF안기단보수구（TprFN ）적우세B세포표위，위심입연구매독다개표위역묘제공의거。방법종GenBank획취T prFN적안기산서렬，채용M obyle、ABCpred화IEDB재선연건종합분석예측TprFN적B세포표위병인공합성다태；표체중조단백 TprFN병경Western blot감정후면역토，획취혈청병측정항체효개；이TprFN면역토혈청、매독환자혈청（설정상인혈청화정상토혈청위음성대조），간접ELISA측정예측적7조인공합성적B세포표위다태적면역반응성화특이성。결과연건종합예측 TprFN적 P1（43‐62AA）、P2（57‐71AA）、P3（81‐88AA）、P4（89‐103AA）、P5（125‐138AA）、P6（231‐251AA）、P7（268‐279AA）가능위B세포표위；표체일가용성단백，WB감정위목적단백，기면역항체효개위1∶12800이상；ELISA결과현시，예측표위P1、P3여TprFN면역토혈청급매독환자혈청균정양성반응，이여대조혈청균불반응。결론 P1、P3위T prF잠재적우세B세포표위。
To predict and identify the dominant B‐cell epitopes of conserved region of Treponema pallidum repeat protein F (TprFN ) and provide the basis for development of polyvalent epitope‐based syphilis vaccine ,the amino acid sequence of TprFN was obtained from GenBank and analyzed with comprehensive meta‐analysis Mobyle ,ABCpred and IEDB online software .The peptides containing predicted epitopes were artificially synthesized . To obtain and measure the titers of antibodies against TprFN ,New Zealand rabbits were immunized with recombinant protein TprFN expressed in E .coli and identified by Western blot (WB) .Sera from TprFN‐immunized rabbits ,syphilis patients ,and normal human and normal rabbits were used to deter‐mine the immunoreactivity and specificity of 7 predicted peptides of TpFN by indirect ELISA .Comprehensive meta‐analysis of online software showed that P1 (43‐62AA) ,P2(57‐71AA) ,P3(81‐88AA) ,P4(89‐103AA) ,P5(125‐138AA) ,P6(231‐251AA) and P7(268‐279AA) might be the B‐cell epitopes .A protein was expressed in a soluble form and identified as TpFN by WB .The ELISA indicated that P1 and P3 were active with TprFN‐immunized rabbit sera and syphilis patient sera but not with negative control sera .These results indicate that P1 and P3 are the potential dominant B‐cell epitopes .