中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2009年
11期
1049-1054
,共6页
假病毒%纽扣珊瑚绿色荧光蛋白%分泌性碱性磷酸酶%中和抗体滴度
假病毒%紐釦珊瑚綠色熒光蛋白%分泌性堿性燐痠酶%中和抗體滴度
가병독%뉴구산호록색형광단백%분비성감성린산매%중화항체적도
Pseudovirus%ZsGreen%SEAP%Neutralizing antibody titer
目的 探讨两种不同的报告基因纽扣珊瑚绿色荧光蛋白(Zoanthus sp.green fluorescent protein,ZsGreen)和分泌性碱性磷酸酶(secreted alkaline phosphatase,SEAP)测量的人乳头状瘤假病毒中和滴度之间的相关性,以及中和滴度和抗体滴度的相关性.方法 将密码子优化的人乳头状瘤病毒(HPV)衣壳蛋白L1、L2基因表达质粒和报告基因质粒共转染293FT细胞,48 h后收集细胞裂解上清,柱层析纯化假病毒,对假病毒滴度进行测定.采集免疫过候选HPV疫苗和Gardasil疫苗的小鼠血清,测量血清的中和滴度和抗体滴度.结果 经统计分析,这两种报告基因系统的假病毒检测的中和滴度结果高度相关(Spearman相关系数r=0.760),而且中和滴度和抗体滴度的相关度很高(Spearman相关系数r=0.577和0.741).结论 两种不同的报告基因ZsGreen和SEAP测量的HPV假病毒中和滴度之间,以及中和滴度和ELISA测定的抗体滴度之间高度相关,揭示了部分HPV疫苗预防病毒入侵的机制,为快速准确鉴定HPV-16和HPV-18候选疫苗的免疫保护效果奠定了基础.
目的 探討兩種不同的報告基因紐釦珊瑚綠色熒光蛋白(Zoanthus sp.green fluorescent protein,ZsGreen)和分泌性堿性燐痠酶(secreted alkaline phosphatase,SEAP)測量的人乳頭狀瘤假病毒中和滴度之間的相關性,以及中和滴度和抗體滴度的相關性.方法 將密碼子優化的人乳頭狀瘤病毒(HPV)衣殼蛋白L1、L2基因錶達質粒和報告基因質粒共轉染293FT細胞,48 h後收集細胞裂解上清,柱層析純化假病毒,對假病毒滴度進行測定.採集免疫過候選HPV疫苗和Gardasil疫苗的小鼠血清,測量血清的中和滴度和抗體滴度.結果 經統計分析,這兩種報告基因繫統的假病毒檢測的中和滴度結果高度相關(Spearman相關繫數r=0.760),而且中和滴度和抗體滴度的相關度很高(Spearman相關繫數r=0.577和0.741).結論 兩種不同的報告基因ZsGreen和SEAP測量的HPV假病毒中和滴度之間,以及中和滴度和ELISA測定的抗體滴度之間高度相關,揭示瞭部分HPV疫苗預防病毒入侵的機製,為快速準確鑒定HPV-16和HPV-18候選疫苗的免疫保護效果奠定瞭基礎.
목적 탐토량충불동적보고기인뉴구산호록색형광단백(Zoanthus sp.green fluorescent protein,ZsGreen)화분비성감성린산매(secreted alkaline phosphatase,SEAP)측량적인유두상류가병독중화적도지간적상관성,이급중화적도화항체적도적상관성.방법 장밀마자우화적인유두상류병독(HPV)의각단백L1、L2기인표체질립화보고기인질립공전염293FT세포,48 h후수집세포렬해상청,주층석순화가병독,대가병독적도진행측정.채집면역과후선HPV역묘화Gardasil역묘적소서혈청,측량혈청적중화적도화항체적도.결과 경통계분석,저량충보고기인계통적가병독검측적중화적도결과고도상관(Spearman상관계수r=0.760),이차중화적도화항체적도적상관도흔고(Spearman상관계수r=0.577화0.741).결론 량충불동적보고기인ZsGreen화SEAP측량적HPV가병독중화적도지간,이급중화적도화ELISA측정적항체적도지간고도상관,게시료부분HPV역묘예방병독입침적궤제,위쾌속준학감정HPV-16화HPV-18후선역묘적면역보호효과전정료기출.
Objective To study the relationship of HPV pseudo-neutralizing titers detected by two different reporter genes: Zoanthus sp. green fluorescent protein (ZsGreen) and secreted alkaline phosphatase (SEAP) , and the relationship between HPV the pseudovirus-neutralizing antibody titer and the antibody titer determined by ELISA method. Methods The plasmids with expression cassettes of the HPV capsid protein L1 and L2 genes after codon optimization and the plasmid with reporter gene (ZsGreen or SEAP) were co-transfected into 293FT cells. The cell lysate supernatants were collected after 48 h culture, then the pseudovirus was purified through POROS column chromatography from the supernatants. After the titer of pseudovirus bulk were measured, HPV-16 and HPV-18 pseudovirus-neutralization assays were carried out for determining the titer of sera collected from immunized mice with HPV candidate vaccine and Gardasil HPV vaccine. Results In statistical analysis, the two reporter gene systems for the detection of the pseudovirus neutralizing antibody titer are highly relevant to each other (Spearman coefficient; r = 0. 760). And their neutralizing antibody titers bear a high degree of correlation with the antibody titer (Spearman coefficient: r= 0.577 and r =0. 741). Conclusion ZsGreen and SEAP pseudovirus neutralizing antibody titers are highly relevant to each other. The neutralizing antibody and the antibody titer are also relevant. These results reveal some mechanism of HPV vaccines to prevent the virus from invading the host cells, and are absolutely useful in the protection efficiency evaluation of the HPV-16 and HPV-18 candidate vaccines.
