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LIBRERIA STUDIUM
Human Cytomegaloviruses
Methods and Protocols
Yurochko, Miller
Editore
Springer
Anno
2014
Pagine
368
ISBN
9781627037877
114,00 €
- Includes cutting-edge methods and protocols
- Provides step-by-step detail essential for reproducible results
- Contains key notes and implementation advice from the experts
With the significant medical role that human cytomegalovirus plays in human disease (from acute disease following primary infection, to chronic disease due to life-long viral persistence), the need to better understand human cytomegalovirus biology and pathogenesis is needed. Human Cytomegaloviruses: Methods and Protocols is designed to be an inclusive document covering all of the techniques and approaches necessary to understand and study the pathobiology of human cytomegalovirus. Topics covered include the history of human cytomegaloviruses, techniques to culture and grow the virus in model cell types, the use of primary cells for the study of human cytomegalovirus pathogenesis, modern molecular techniques for assessing the biological consequences of viral infection, animal models for study of cytomegalovirus replication and numerous other topics of current interest. Written in the successful Methods in Molecular Biology series format, each chapter includes an introduction, list of necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and easily accessible, Human Cytomegaloviruses: Methods and Protocols serves as a tool for basic scientists as well as clinical scientists with an interest in the basic fundamental aspects of viral gene expression and specific aspects of viral pathogenesis.
Contents
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix
1 History of the Molecular Biology of Cytomegaloviruses . . . . . . . . . . . . . . . . . 1
Mark F. Stinski
2 Overview of Human Cytomegalovirus Pathogenesis . . . . . . . . . . . . . . . . . . . . 15
Maciej T. Nogalski, Donna Collins-McMillen,
and Andrew D. Yurochko
3 Distinct Properties of Human Cytomegalovirus Strains
and the Appropriate Choice of Strains for Particular Studies . . . . . . . . . . . . . . 29
Giada Frascaroli and Christian Sinzger
4 Use of Diploid Human Fibroblasts as a Model System to Culture,
Grow, and Study Human Cytomegalovirus Infection. . . . . . . . . . . . . . . . . . . . 47
Elizabeth A. Fortunato
5 Use of Recombinant Approaches to Construct Human
Cytomegalovirus Mutants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
Iryna Dekhtiarenko, Luka Čičin-Šain, and Martin Messerle
6 The Use of Primary Human Cells (Fibroblasts, Monocytes, and Others)
to Assess Human Cytomegalovirus Function . . . . . . . . . . . . . . . . . . . . . . . . . . 81
Emma Poole, Matthew Reeves, and John H. Sinclair
7 Hematopoietic Long-Term Culture (hLTC)
for Human Cytomegalovirus Latency and Reactivation . . . . . . . . . . . . . . . . . . 99
Mahadevaiah Umashankar and Felicia Goodrum
8 Analysis of Cytomegalovirus Binding/Entry-Mediated Events. . . . . . . . . . . . . 113
Gary C.T. Chan and Andrew D. Yurochko
9 Use of 5-Ethynyl-2′-Deoxyuridine Labelling and Flow Cytometry
to Study Cell Cycle-Dependent Regulation of Human Cytomegalovirus
Gene Expression . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
Lüder Wiebusch and Christian Hagemeier
10 Methods for Studying the Function of Cytomegalovirus GPCRs . . . . . . . . . . . 133
Christine M. O’Connor and William E. Miller
11 Methods for the Detection of Cytomegalovirus
in Glioblastoma Cells and Tissues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165
Charles S. Cobbs, Lisa Matlaf, and Lualhati E. Harkins
12 Methods to Study the Nucleocytoplasmic Transport of Macromolecules
with Respect to Their Impact on the Regulation
of Human Cytomegalovirus Gene Expression . . . . . . . . . . . . . . . . . . . . . . . . . 197
Marco Thomas, Barbara Zielke, Nina Reuter, and Thomas Stamminger
13 Fluorescence-Based Laser Capture Microscopy
Technology Facilitates Identification of Critical
In Vivo Cytomegalovirus Transcriptional Programs . . . . . . . . . . . . . . . . . . . . . 217
Craig N. Kreklywich, Patricia P. Smith, Carmen Baca Jones,
Anda Cornea, Susan L. Orloff, and Daniel N. Streblow
14 Techniques for Characterizing Cytomegalovirus-Encoded miRNAs . . . . . . . . . 239
Lauren M. Hook, Igor Landais, Meaghan H. Hancock,
and Jay A. Nelson
15 What We Have Learned from Animal Models of HCMV. . . . . . . . . . . . . . . . . 267
Pranay Dogra and Tim E. Sparer
16 Rodent Models of Congenital Cytomegalovirus Infection . . . . . . . . . . . . . . . . 289
Djurdjica Cekinovic, Vanda Juranic Lisnic, and Stipan Jonjic
17 Recent Approaches and Strategies in the Generation
of Antihuman Cytomegalovirus Vaccines. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 311
Suresh B. Boppana and William J. Britt
18 Approaches for the Generation of New Anti- cytomegalovirus Agents:
Identification of Protein–Protein Interaction Inhibitors
and Compounds Against the HCMV IE2 Protein . . . . . . . . . . . . . . . . . . . . . . 349
Beatrice Mercorelli, Giorgio Gribaudo, Giorgio Palù,
and Arianna Loregian
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 365
