Gary P. Wormser, Section Editor Human Papillomaviruses: Methods and Protocols Edited by Clare Davy and John Doorbar Totowa, NJ: Humana Press, 2005. 512 pp., illustrated. $145.00 (cloth)

Significant advances have been made in the past decade in the field of genital cancer and human papillomavirus (HPV) research. In contrast to most viruses, HPVs cannot be easily propagated in vitro. Moreover, these viruses exhibit strong host and tissue tropism. Specialized techniques have been developed to overcome these limitations. This monograph, which is part of the Methods in Molecular Medicine series, focuses on some of the methods that have enabled the scientific community to comprehensively study HPV infection in vitro and in vivo. Since our current knowledge of diseases caused by HPV is derived in part from our ability to study HPV infection in humans, several of the methods proposed in the book can be applied to clinical samples. Overall, this book provides the reader with detailed protocols that cover a wide spectrum of novel and well-established techniques applicable to HPV research. This variety of techniques will be appealing to basic as well as clinical scientists involved in cancer research. A diverse team of 79 contributors, all experts in their field, present an authoritative and comprehensive text that describes cutting-edge molecular biology methods for HPV research. Each of the 33 chapters is well written and well referenced. The book is not formally divided into sections; however, the chapters belong to 3 distinct themes that encompass a broad range of techniques: the detection and characterization of HPV DNA or mRNA, the analysis of the life cycle of HPVs, and the production and functional analysis of HPV proteins. Some of these protocols (such as those for HPV DNA detection and typing) are well established, while others (such as real-time PCR assays and HPV pseudovirions) are novel techniques. Each chapter provides a summary; a short, helpful introduction on the state of knowledge of a research topic; a Materials section for each step of the different techniques that enumerates the instruments and reagents required; a Methods section that describes the procedures step-by-step; and, finally, a section entitled “Notes,” in which the authors discuss practical issues related to these techniques. Each chapter contains useful tables for primer sequences or protocols, as well as schematic drawings of the methods or figures of results obtained with the procedures described. The tables and figures stand on their own and complement the text. Molecular typing methods have been instrumental in delineating the epidemiology of HPV infection. The first chapters focus on HPV DNA detection methods, as well as the identification of HPV types or variants. The most widely used PCR assays for genital and cutaneous HPV types are discussed along with various processing methods for a variety of samples screened for HPV. The histochemical analysis of biopsy samples is presented along with the use of potential biomarkers for the progression of cervical intraepithelial neoplasia. The analysis of HPV expression in vivo is also discussed in several sections. These chapters are followed by protocols for establishing keratinocyte cell lines or cell lines derived from HPV-induced lesions with a description of organotypic culturing techniques and retrovirus gene transfer to analyze HPV gene regulation. These tools are useful for studying the HPV life cycle, HPV gene regulation, and protein functions. Several approaches for the investigation of HPV protein functions, including the analysis of interactions between HPV and cellular proteins (e.g., E7/Rb and p53/E6), transformation assays, and the induction of apoptosis by HPV proteins, are detailed in the following chapters. There are also chapters on 2 animal models of HPV pathogenesis: the severe combined immune deficiency mouse and the cotton rabbit. Several strategies for analyzing HPV expression at the level of mRNA or viral proteins are presented in the following chapters. The chapter on the investigation of viral transcription regulation proposes 3 methods: DNase footprinting, electrophoretic mobility shift assay, and bisulfite sequencing procedures. Finally, methods to generate HPV-specific viruslike particles are presented. This book is much more than a molecular biology cookbook or a collection of protocols. Investigators new to this field and well-established researchers alike will find precious information in the “Notes” sections, often derived from the experience of the authors, that is unavailable from other sources. The “Notes” sections contain advice that could only have been obtained by spending some time in the laboratory with the authors. Most of the information provided in these sections throughout the book is very useful, especially when potential pitfalls are discussed. Use of caution during the analysis of results is also addressed. Some useful Web site addresses are provided. The Protocol section describes, in quite a bit of detail, experimental procedures on the topics presented. The book also succeeds in giving a variety of protocols, from histopathological examination to HPV DNA detection and HPV-cellular protein interactions. Interestingly, the same techniques are often discussed in different chapters for different purposes by different investigators, each with a different view on their value or application. The authors usually refer to other chapters when the technique is discussed in more than one chapter. This is a comprehensive and invaluable resource on the technical aspects of HPV research. We should congratulate the editors for assembling such an expert team that has produced this solid information. It will become an important resource for those who work in the realm of HPV research, including clinical pathologists, clinical microbiologists, virologists, and infectious disease specialists, as well as their trainees. Potential conflicts of interest. F.C. has received reagents for HPV research from Roche Molecular Systems.

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