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
2008年
11期
984-988
,共5页
张晶晶%杨光%杨春%蒿艳蓉%曹骥%欧超%李瑗%班克臣%Hubert E.Blum%苏建家
張晶晶%楊光%楊春%蒿豔蓉%曹驥%歐超%李瑗%班剋臣%Hubert E.Blum%囌建傢
장정정%양광%양춘%호염용%조기%구초%리원%반극신%Hubert E.Blum%소건가
乙肝病毒%感染%树鼩%原代肝细胞
乙肝病毒%感染%樹鼩%原代肝細胞
을간병독%감염%수구%원대간세포
Hepatitis B virus%Infection%Tupala%Primary hepatocytes
目的 为进一步研究人类乙肝病毒(HBV)提供接近自然感染状态的理想细胞模型.方法 体外二步灌流法分离树鼩原代肝细胞,纯化后的乙肝患者血清感染上述肝细胞,Southern blot和Noahem blot检测感染后细胞内的DNA和RNA,ELISA方法 检测细胞上清的HBsAg,免疫组化检测细胞内HBsAg的表达.结果 可检测出肝细胞内cccDNA(共价闭合环状DNA)、pgRNA(前基因组RNA)和sgRNA(亚基因组mRNA),感染后第7天,信号开始增强,持续到实验结束的第14天.细胞上清中HBsAg自第1天到第5天S/CO值逐渐下降,随后S/CO值逐渐升高.结论 HBV可在原代树鼩肝细胞中复制和表达.
目的 為進一步研究人類乙肝病毒(HBV)提供接近自然感染狀態的理想細胞模型.方法 體外二步灌流法分離樹鼩原代肝細胞,純化後的乙肝患者血清感染上述肝細胞,Southern blot和Noahem blot檢測感染後細胞內的DNA和RNA,ELISA方法 檢測細胞上清的HBsAg,免疫組化檢測細胞內HBsAg的錶達.結果 可檢測齣肝細胞內cccDNA(共價閉閤環狀DNA)、pgRNA(前基因組RNA)和sgRNA(亞基因組mRNA),感染後第7天,信號開始增彊,持續到實驗結束的第14天.細胞上清中HBsAg自第1天到第5天S/CO值逐漸下降,隨後S/CO值逐漸升高.結論 HBV可在原代樹鼩肝細胞中複製和錶達.
목적 위진일보연구인류을간병독(HBV)제공접근자연감염상태적이상세포모형.방법 체외이보관류법분리수구원대간세포,순화후적을간환자혈청감염상술간세포,Southern blot화Noahem blot검측감염후세포내적DNA화RNA,ELISA방법 검측세포상청적HBsAg,면역조화검측세포내HBsAg적표체.결과 가검측출간세포내cccDNA(공개폐합배상DNA)、pgRNA(전기인조RNA)화sgRNA(아기인조mRNA),감염후제7천,신호개시증강,지속도실험결속적제14천.세포상청중HBsAg자제1천도제5천S/CO치축점하강,수후S/CO치축점승고.결론 HBV가재원대수구간세포중복제화표체.
Objective To provide a better cell model of closely nature infectious state for further research of hepatitis B virus(HBV). Methods Primary tupaia hepatocytes were isolated by the two-step perfusion method. The hepatocytes were then infected with purified serum from patients with hepatitis B. DNA and RNA isolated from the hepatocytes were detected with Southern blot and Northern blot. HBsAg in supernatant was tested by immunohistochemical method. Results cccDNA, pgRNA and sgRNA could be detected by Southern blot and Northem blot, and strong signals could be seen from day 7 to day 14 post-in-fection. The S/CO value of HBsAg in supernatant decreased from day 1 to day 5 and then increased after 5 day. Conclusion Primary tupaia hepatocytes are competent for infection with HBV. HBV can stably repli-cate and express in HBV-infected tupaia hepatocytes.
