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A cutting-edge collection of basic and state-of-the-art methods optimized for investigating the molecular biology of this class of retrovirus. These readily reproducible techniques range from methods for the isolation and detection of human retroviruses to cutting-edge methods for exploring the interplay between the viruses and the host. Here, the researcher will find up-to-date techniques for the isolation and propagation of HIV, HTLV, and foamy virus from a variety of sources. There are also assays for determining the cell tropism of HIV-1, the coreceptor usage of HIV-1, and human gene expression with HIV-1 infection by microarrays, as well as for phenotyping HIV-1 infected monocytes and examining their fitness. Highlights include the detection and quantification of HIV-1 in resting CD4+, a new cloning system for making recombinent virus, cDNA microarrays, and the determination of genetic polymorphisms in two recently identified HIV-1 co-factors that are critical for HIV-1 infection.
A comprehensive compilation of research techniques necessary for investigating the virology, immunology and molecular biology of HIV-1. Protocols are also provided which represent state of the art approaches to a wide spectrum of HIV related issues.
The worldwide impact of infection with human immunodeficiency virus type 1 (HIV- is reflected in the cumulative number ofHIV- 1 infections, which is now predicted to exceed 40 million by the year 2000---equivalent to the n- ber of humans who perished in World War II. The medical and scientific - sponse to the HIV-1 pandemic has steadily grown since its recognition in 1981. The outlay by the United States alone for HIV research funded by the National Institutes of Health in 1997 was $1. 4 billion. Laboratory-based HIV research has brought together academic clinicians, retrovirologists, molecular biologists, and immunologists in the formation of research teams attempting to dissect the viral and host factors contributing to disease pathogenesis. Increasing focus is being placed on those aspects of viral biology and host immune responses that bear on the development of vaccines to prevent HIV infection. HIVProtocols reflects the state of HIV research in several ways. First, chapters are organized into four sections: Virology, Molecular Biology, Humoral Immunology, and Cellular Immunology. This organization is a natural consequence of the diverse scientific disciplines that have been attracted to HIV research. Second, the chapters reflect such diverse research directions as viral coreceptor usage, quantitation of viral genomes, HIV promoter function, B-cell epitope mapping, and measurements of T-cell function, each of which bears on the goal of understanding the viral and host immune responses that will be critical to the design of effective preventive vaccines.