CAJ | 학술논문

目的：预测并鉴定沙眼衣原体（Ct）Tarp蛋白的B细胞表位，为进一步研究Tarp蛋白的生物学特性及表位疫苗研制奠定基础。方法：利用生物信息学软件综合分析、预测筛选E血清型CtTarp蛋白的B细胞表位，获得6个潜在目的表位。将人工合成的表位肽段与弗氏佐剂充分乳化，经背部皮下多点注射途径免疫BALB/c小鼠（每次100μg/只，3只/组，共24只），间隔2周，共免疫3次。采用ELISA法检测免疫小鼠血清中Tarp蛋白特异性抗体IgG及其亚类IgG1、IgG2a，以及阴道分泌物中Tarp蛋白特异性sIgA抗体；进一步采用间接免疫荧光实验、免疫沉淀实验和Westernblot法检测表位免疫的血清抗体与Tarp蛋白结合的特异性。结果：筛选并鉴定了Tarp蛋白的一个免疫优势B细胞表位（aa171-186），该表位可以刺激小鼠产生较高水平的Tarp蛋白特异性IgG抗体和sIgA抗体，且抗体亚类以IgG1为主。免疫荧光实验、免疫沉淀实验和Westernblot法检测结果显示，该段表位肽免疫血清抗体可特异性识别Tarp蛋白。结论：获得了CtTarp蛋白的一个免疫优势B细胞表位，为进一步研究Tarp蛋白的生物学特性及表位疫苗研制奠定基础。
목적：예측병감정사안의원체（Ct）Tarp단백적B세포표위，위진일보연구Tarp단백적생물학특성급표위역묘연제전정기출。방법：이용생물신식학연건종합분석、예측사선E혈청형CtTarp단백적B세포표위，획득6개잠재목적표위。장인공합성적표위태단여불씨좌제충분유화，경배부피하다점주사도경면역BALB/c소서（매차100μg/지，3지/조，공24지），간격2주，공면역3차。채용ELISA법검측면역소서혈청중Tarp단백특이성항체IgG급기아류IgG1、IgG2a，이급음도분비물중Tarp단백특이성sIgA항체；진일보채용간접면역형광실험、면역침정실험화Westernblot법검측표위면역적혈청항체여Tarp단백결합적특이성。결과：사선병감정료Tarp단백적일개면역우세B세포표위（aa171-186），해표위가이자격소서산생교고수평적Tarp단백특이성IgG항체화sIgA항체，차항체아류이IgG1위주。면역형광실험、면역침정실험화Westernblot법검측결과현시，해단표위태면역혈청항체가특이성식별Tarp단백。결론：획득료CtTarp단백적일개면역우세B세포표위，위진일보연구Tarp단백적생물학특성급표위역묘연제전정기출。
Objective: To predict and identify the B-cell epitope ofChlamydia trachomatis Tarp.Methods:The amino acid sequence of Tarp was analyzed using computer-assisted techniques to scan B-cell epitopes, and 6 possible linear B-cell epitopes peptides (aa80-95, aa107-123, aa152-170, aa171-186, aa239-253 and aa497-513) with high predicted antigenicity and high conservation were investigated. After coupling to KLH, the syn-thetic peptides were emulsiifeld with Freund’s adjuvant. Then, BALB/c mice were randomly assigned to receive (subcutaneous injection) peptides-KLH, KLH or PBS (n=3, 100 μg/time/mouse), respectively, and the same immunization schedule was repeated third times at a 2-week interval. ELISA was used to detect the IgG and sub-types IgG1, IgG2a in the serum as well as the sIgA in the vaginal secretions. Furthermore, the speciifc binding of anti-sera produced by epitopes immunization to natural Tarp antigen was detected with indirect immunolfuo-rescence assay (IFA), immunoprecipitated and Western blot analysis, respectively.Results: The epitope peptide F4 (aa171-186) was able to induce signiifcant Tarp speciifc antibody IgG in serum and sIgA in vaginal wash as compared with KLH or PBS (1:100) (P<0.05), and the major IgG antibody subtype was IgG1. Western blot and IFA results conifrmed that speciifc antibodies were induced by epitope aa171-186 recognized natural Tarp anti-gen.Conclusion: Our results demonstrated that the predictive epitopes (aa171-186) are immunodominant B-cell epitope of Tarp, and if proven protective and safe, in conjunction with others well-documented epitopes, they may be included into a candidate epitope-based vaccine.