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(American Journal of Pathology. 2004;165:707-718.)
© 2004 American Society for Investigative Pathology

Technical Advance

Highly Tissue Substructure-Specific Effects of Human Papilloma Virus in Mucosa of HIV-Infected Patients Revealed by Laser-Dissection Microscopy-Assisted Gene Expression Profiling

Nicole Baumgarth^*, Richard Szubin, Greg M. Dolganov, Mitchell R. Watnik, Deborah Greenspan, Maria Da Costa, Joel M. Palefsky, Richard Jordan, Mario Roederer^¶ and John S. Greenspan

From the Center for Comparative Medicine^* and the Department of Statistics, University of California, Davis, California; the Department of Stomatology, and the Division of Pulmonary and Critical Care Medicine, Department of Medicine, University of California, San Francisco, California; and the Vaccine Research Center,^¶ National Institutes of Health, Bethesda, Maryland

Human papilloma virus (HPV) causes focal infections of epithelial layers in skin and mucosa. HIV-infected patients on highly active antiretroviral therapy (HAART) appear to be at increased risk of developing HPV-induced oral warts. To identify the mechanisms that allow long-term infection of oral epithelial cells in these patients, we used a combination of laser-dissection microscopy (LDM) and highly sensitive and quantitative, non-biased, two-step multiplex real-time RT-PCR to study pathogen-induced alterations of specific tissue subcompartments. Expression of 166 genes was compared in three distinct epithelial and subepithelial compartments isolated from biopsies of normal mucosa from HIV-infected and non-infected patients and of HPV32-induced oral warts from HIV-infected patients. In contrast to the underlying HIV infection and/or HAART, which did not significantly elaborate tissue substructure-specific effects, changes in oral warts were strongly tissue substructure-specific. HPV 32 seems to establish infection by selectively enhancing epithelial cell growth and differentiation in the stratum spinosum and to evade the immune system by actively suppressing inflammatory responses in adjacent underlying tissues. With this highly sensitive and quantitative method tissue-specific expression of hundreds of genes can be studied simultaneously in a few cells. Because of its large dynamic measurement range it could also become a method of choice to confirm and better quantify results obtained by microarray analysis.

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