中华皮肤科杂志

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人乳头瘤病毒6/11型体外感染HaCaT细胞的研究

吴剑波, 李新宇, 郑家润   

  1. 中国医学科学院、中国协和医科大学皮肤病研究所 南京 210042
  • 收稿日期:2005-05-14 出版日期:2006-04-15 发布日期:2006-04-15

In vitro infection of HaCaT cells by HPV6/11

WU Jian-bo, LI Xin-yu, ZHENG Jia-run   

  1. Institute of Dermatology, Chinese Academy of Medical Sciences & Peking Union Medical College, Nanjing 210042, China
  • Received:2005-05-14 Online:2006-04-15 Published:2006-04-15

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摘要： 目的 研究人乳头瘤病毒(HPV)6/11型病毒颗粒体外感染HaCaT细胞后的感染标志表达及检测方法。方法 从临床尖锐湿疣标本中提取HPV6/11病毒颗粒,用于感染单层培养的HaCaT细胞,以FQ-PCR监测病毒悬液与感染后HaCaT细胞的病毒DNA载量;用引物原位延伸标记方法原位检测感染后HaCaT细胞中HPV DNA的存在及分布;以免疫组化法检测HPV的衣壳抗原表达。结果 在病毒悬液中HPV DNA达到10~5拷贝/ml以上时,病毒处理后的HaCaT细胞FQ-PCR检测结果呈稳定阳性;PRINS方法可检测到呈灶状分布的胞核阳性染色的细胞;免疫组化法未能检测到HPV衣壳蛋白的表达。结论 一定浓度的HPV6/11可以在体外培养系统中感染HaCaT细胞,在其细胞核内出现HPV DNA,但无衣壳蛋白表达。

关键词: 乳头状瘤病毒, 人, 体外研究, HaCaT细胞株

Abstract: Objective To investigate the expression of infectious markers of HaCaT cells infected in vitro by HPV 6/11.Methods HPV 6/11 virions isolated from condyloma acuminata lesions were used to infect cultured HaCaT cells.HPV DNA was determined quantitatively by FQ-PCR and primed in situ labeling (PRINS).The HPV capsid antigen was detected by immunohistochemistry method.Results HPV DNA was consistently positive in HaCaT cell culture,when DNA copy number was over 10~5/mL in suspension.A focal distribution of HPV DNA(+) was revealed in HaCaT cells.Capsid antigen was not found in infected HaCaT cells.Conelusion It is suggested that HPV6/11 could infect HaCaT cells in vitro.HPV DNA could be found in the nuclei of infected HaCaT cells,but capsid protein is not expressed.

Key words: Papillomavirus,human, in vitro, HaCaT cell strain