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Techniques in HIV research
Cell, Vol. 65. 1103-l 105, June 29, 1991, Copyright 0 1991 by Cell Press
Book Reviews
A to Z of Methods in HIV Research Techniques in HIV Research. Edited by A. Aldovini and B. D. Walker. New York: Stockton Press. (1990). 285 pp. $49.95.
HIV research nowadays represents the achievement of one of the greatest world-wide mobilizations of the scientific community during the last decades. Only 8 years after its discovery, the number of investigations on HIV and on the disease that it causes is still increasing. Our current knowledge of HIV is the result of the work of numerous experts from different fields including virology, medicine, biochemistry, molecular biology, immunology, and epidemiology. Such an overflow of scientific interests urges the investigators to seek a general knowledge in different disciplines in order to understand and to perform different techniques efficiently. Consequently, a laboratory manual on techniques used in HIV research is almost a “must” for any laboratory working on HIV-related research projects. Fortunately, this manual edited by Aldovini and Walker provides almost all the information that one might require to appreciate the state of the game. The book is fairly comprehensive. It contains four main sections on techniques used in viral diagnostics, basic virology, molecular biology, immunology, and finally a section on guidelines to evaluate potential candidates as anti-HIV agents in vitro. Each section has several chapters written by specialists in each field who present detailed methods and also provide a troubleshooting guide for the inexperienced researcher. In the section dealing with methods in viral diagnostics one can find all the techniques used for HIV antibody detection in serum, assay of HIV protein p24/p25 in serum, and HIV nucleic acid (RNA and DNA) detection in clinical specimens. This section should satisfy both the inexperienced investigator who has to set up a laboratory for HIV diagnosis and also the experienced researcher who might require some additional information on one of the techniques. The section on basic virology techniques provides information and protocols for isolation of viruses from T lymphocytes, monocytes, and their derivatives present in various patient tissues. In addition, one can find details on virus infection protocol cultures that are employed for the production of high titers of HIV. Quantitative methods to estimate virus infectivity and to determine the presence of the virus itself have been presented using protocols based on the assay of the cytopathic effect and syncytium formation in infected cultures, the indirect immunofluorescence assay to demonstrate the number of infected cells, the reverse transcriptase assay to show the activity of the viral enzyme that will synthesize the provirus, and lastly the polymerase chain reaction (PCR) assay to illustrate the presence of HIV nucleic acids.
The PCR assay, in contrast to the former assays, should be manipulated in a more specialized laboratory because of the high risk of contamination. The third section of this book, which is on techniques in molecular biology, is a short review of the approach used to clone HIV proviral DNA and transfection of molecularly cloned HIV genomes. This latter section is concise since one can find the detailed methods in more specialized manuals dealing with techniques in molecular biology. Cf special interest is the section focusing on neutralizing antibodies and the cellular immune response to HIV. For the detection of neutralizing antibodies, this manual provides a sensitive, rapid, and practical method that should be used by the different investigators in the field as a standard reference neutralization assay. Two chapters, devoted to HIV-specific cytotoxicity, give efficient techniques to discriminate between the two predominant anti-HIV cytolytic reactivities: cytotoxic T cells and antibodydependent cellular cytotoxicities, CTL and ADCC. The last chapter on virusspecific host immune response discusses the T helper cell responses that are required for optimal cytotoxic T cell response to kill virus-infected cells as well as for antibody responses to protein antigens. T helper cell activity is measured by antigen-induced proliferation or by interleukin-2 production assays. From the different sections, the editors have succeeded in forming a coherent picture of techniques used in HIV research. The editors finish the manual with appendices giving useful information on World Health Organization centers and the source of reagents used in HIV research. They also provide safety guidelines for investigators working with HIV and recombinant vaccinia viruses. In conclusion, this is the type of book that should be handy to almost anyone working on HIV. The techniques and methods are clearly presented and, for further details, a selected list of references is presented on each topic. Perhaps in a few years, such a manual should be reedited to include new findings and thus polish the methods mentioned here. Whatever is the case, this book certainly will join the series of some laboratory manual classics. Ara G. Hovanessian, Yves RivYn?, and Luc Montagnier Department of AIDS and Retroviruses lnstitut Pasteur 28 Rue du Dr. Roux 75024 Paris France
All the King’s Men Peptide Growth Factors and Their Receptors. Handbook of Pharmacology. Volume 95. Parts 1 and 2. Edited by M. B. Sporn and A. B. Roberts. Heidelberg: Springer-Verlag. (1990). 794 pp (Part 1) and 727 pp (Part 2). DM 498.
Book Reviews
A to Z of Methods in HIV Research Techniques in HIV Research. Edited by A. Aldovini and B. D. Walker. New York: Stockton Press. (1990). 285 pp. $49.95.
HIV research nowadays represents the achievement of one of the greatest world-wide mobilizations of the scientific community during the last decades. Only 8 years after its discovery, the number of investigations on HIV and on the disease that it causes is still increasing. Our current knowledge of HIV is the result of the work of numerous experts from different fields including virology, medicine, biochemistry, molecular biology, immunology, and epidemiology. Such an overflow of scientific interests urges the investigators to seek a general knowledge in different disciplines in order to understand and to perform different techniques efficiently. Consequently, a laboratory manual on techniques used in HIV research is almost a “must” for any laboratory working on HIV-related research projects. Fortunately, this manual edited by Aldovini and Walker provides almost all the information that one might require to appreciate the state of the game. The book is fairly comprehensive. It contains four main sections on techniques used in viral diagnostics, basic virology, molecular biology, immunology, and finally a section on guidelines to evaluate potential candidates as anti-HIV agents in vitro. Each section has several chapters written by specialists in each field who present detailed methods and also provide a troubleshooting guide for the inexperienced researcher. In the section dealing with methods in viral diagnostics one can find all the techniques used for HIV antibody detection in serum, assay of HIV protein p24/p25 in serum, and HIV nucleic acid (RNA and DNA) detection in clinical specimens. This section should satisfy both the inexperienced investigator who has to set up a laboratory for HIV diagnosis and also the experienced researcher who might require some additional information on one of the techniques. The section on basic virology techniques provides information and protocols for isolation of viruses from T lymphocytes, monocytes, and their derivatives present in various patient tissues. In addition, one can find details on virus infection protocol cultures that are employed for the production of high titers of HIV. Quantitative methods to estimate virus infectivity and to determine the presence of the virus itself have been presented using protocols based on the assay of the cytopathic effect and syncytium formation in infected cultures, the indirect immunofluorescence assay to demonstrate the number of infected cells, the reverse transcriptase assay to show the activity of the viral enzyme that will synthesize the provirus, and lastly the polymerase chain reaction (PCR) assay to illustrate the presence of HIV nucleic acids.
The PCR assay, in contrast to the former assays, should be manipulated in a more specialized laboratory because of the high risk of contamination. The third section of this book, which is on techniques in molecular biology, is a short review of the approach used to clone HIV proviral DNA and transfection of molecularly cloned HIV genomes. This latter section is concise since one can find the detailed methods in more specialized manuals dealing with techniques in molecular biology. Cf special interest is the section focusing on neutralizing antibodies and the cellular immune response to HIV. For the detection of neutralizing antibodies, this manual provides a sensitive, rapid, and practical method that should be used by the different investigators in the field as a standard reference neutralization assay. Two chapters, devoted to HIV-specific cytotoxicity, give efficient techniques to discriminate between the two predominant anti-HIV cytolytic reactivities: cytotoxic T cells and antibodydependent cellular cytotoxicities, CTL and ADCC. The last chapter on virusspecific host immune response discusses the T helper cell responses that are required for optimal cytotoxic T cell response to kill virus-infected cells as well as for antibody responses to protein antigens. T helper cell activity is measured by antigen-induced proliferation or by interleukin-2 production assays. From the different sections, the editors have succeeded in forming a coherent picture of techniques used in HIV research. The editors finish the manual with appendices giving useful information on World Health Organization centers and the source of reagents used in HIV research. They also provide safety guidelines for investigators working with HIV and recombinant vaccinia viruses. In conclusion, this is the type of book that should be handy to almost anyone working on HIV. The techniques and methods are clearly presented and, for further details, a selected list of references is presented on each topic. Perhaps in a few years, such a manual should be reedited to include new findings and thus polish the methods mentioned here. Whatever is the case, this book certainly will join the series of some laboratory manual classics. Ara G. Hovanessian, Yves RivYn?, and Luc Montagnier Department of AIDS and Retroviruses lnstitut Pasteur 28 Rue du Dr. Roux 75024 Paris France
All the King’s Men Peptide Growth Factors and Their Receptors. Handbook of Pharmacology. Volume 95. Parts 1 and 2. Edited by M. B. Sporn and A. B. Roberts. Heidelberg: Springer-Verlag. (1990). 794 pp (Part 1) and 727 pp (Part 2). DM 498.