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In vitro HPV-11 infection of human foreskin
L H Smith1, C Foster, M E Hitchcock
1Department of Obstetrics and Gynecology, University of California at Davis, School of Medicine 95816.
Insights
Researchers developed an in vitro method to detect human papillomavirus type 11 (HPV-11) infection in keratinocytes. This new system allows for the study of early HPV-11 infection events.
Area of Science:
- Virology
- Dermatology
- Molecular Biology
Background:
- Human papillomavirus type 11 (HPV-11) infection is studied using xenografts of infected neonatal human foreskin epithelium in athymic nude mice.
- Previous models have limitations in studying the early infectious process.
Purpose of the Study:
- To establish and validate an in vitro system for detecting HPV-11 infection in cultured keratinocytes.
- To analyze the expression of early HPV-11 transcripts in vitro.
Main Methods:
- Neonatal human foreskin keratinocytes were infected with HPV-11 in vitro.
- Polymerase chain reaction (PCR) was used with specific primers to detect HPV-11 E1^E4 mRNA.
- Heat inactivation of the inoculum was performed to assess its effect on infection.
Main Results:
- HPV-11 infection was detected in cultured keratinocytes using PCR.
- Expression of the HPV-11 E1^E4 mRNA was confirmed in cultured cells.
- Heat inactivation of the inoculum abrogated E1^E4 mRNA expression.
- Infected explants and subsequent passages of keratinocytes continued to produce E1^E4 mRNA for up to 5 weeks.
Conclusions:
- An effective in vitro system for studying HPV-11 infection has been developed.
- This system allows for the detection of HPV-11 infection and early gene expression in cultured keratinocytes.
- The in vitro model offers a novel approach for investigating the initial stages of HPV-11 infection.
Abstract:
Study of the infectious process of human papillomavirus type 11 (HPV-11) has been facilitated by the discovery that HPV-11-infected neonatal human foreskin epithelium can proliferate as xenografts into condyloma-like growths within athymic nude mice. Here we describe detection of HPV-11 infection of neonatal human foreskin-derived keratinocytes, infected and cultured entirely in vitro, by use of the polymerase chain reaction and primers straddling the splice donor/acceptor site of the most prevalent early gene HPV-11 transcript (E1 increase E4). Expression of the E1 increase E4 HPV-11 mRNA is abrogated by 60 degrees C heat inactivation of the inoculum. HPV-11-infected foreskin explants continue to produce the E1 increase E4 mRNA for up to 5 weeks in culture, and second-passage keratinocytes derived from infected explant outgrowths continue to produce the E1 increase E4 mRNA. The in vitro system described here provides a new way to study HPV-11 infection and may be useful in evaluating early events of infection.