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A cook book of immunological methods. Antibodies. A laboratory manual
Molecular Immunology, Vol. 28, No. 6, pp. 681682, Pergamon Press plc. Printed in Great Britain.
1991
BOOK REVIEWS
The Scid Mouse, Characterization and Potential Uses. Edited bv M. J. Bosma. R. A. Phillins and W. Schuler. Current T&s in Microbklogy and inukology, Vol. 152. Springer, Berlin, 1989, 263 pp. DMI32. This comprehensive book presents data on severe combined immunodeflcient (scid) mice. It provides their background and characterization, including cellular and molecular aspects. It reveals the many areas in immunology in which these mice can serve as useful experimental models, which allow studying questions that were heretofore unapproachable. The scid mice were discovered by Bosma in 1980. He was performing a routine screening of immunoglobulin allotypes in the heavy chain congenic BALB/c and C.B-17 mouse strains, when he noticed that some of the Cb-17 mice did not express serum immunoglobulin of the appropriate ahotype. Further checking revealed that the mice actually lacked all of the major immunoglobulin classes, and it became apparent that these were mutant mice. By 1987, the uniqueness and versatility of the scid mouse model was well recognized. At that time the European Molecular Biology Organi~tion (EMBO) held a workshop on the subject and this book is a collection of the papers presented thereat. The book is impressive in its scope, covering the full range of important research issues that have been investigated on these mice. Data provided by Bosma point out the developmental stages in the T and B lineages at which differentiation ceases. The mutation was found to interrupt the process of gene-rearrangement, due to a defective V(D)J recombinase system in developing lymphoid cells. It is manifested in prevention of the proper expression of antigen receptors on both B and T lymphocytes. The scid mice thus offer a special expe~mental model for studies of lymphoid development. A detailed study on hemopoiesis in the scid mouse is reported by Phillip and Fullop, showing normal development of myeloid and NK cells and absence of mature lymphoid elements. These studies led also to the observation on increased radiation sensitivity in these mice, which is of particular interest in relation to Ataxia telangiectasia in the human. Extensive analysis of B and T lymphocyte development are reported from several laboratories. Noteworthy examples are Hardy et ~1,“sexperiments illustrating sequential stages of development within the bone marrow B cell compartment by dete~ination of cell membrane phenotypes, and von Boehmer’s analysis of T lymphocyte development, which makes use of the scid along with mice transgenic for the T cell receptor (TCR), which contain productively arranged alpha and beta TCR genes, and of crosses between these mice. A considerable volume of data relates to potential uses for the scid mice. They include studies on the transplantation of lymphohemopoietic cells from normal mice to examine reconstitution of the different cellular elements in intact and irradiated recipients, on grafting of bone marrow and fetal liver cells, and of fetal primordia, for examination of lymphoid developments and studies of myeloid and lymphoid stem cells. The studies on bone marrow transplantation, focussing on mechanisms underlying graft rejection, further emphasize the advantages of these mice. A study by Bennet et al. examines extensively the histocompatibility differences
identified by the scid mice, and analyzes the cellular mechanisms involved. Since graft rejection in these mice cannot be related to either T or B cells, the cause of bone marrow rejection may be NK cells, which are functional in the scid mice. Other particularly interesting potential uses include engraftment of human cells. The idea is to devise in viuo culture conditions that will allow studies on isolated human cells. Once this is achieved, a whole new spectrum of studies will be feasible. The part of the book devoted to this area includes research from different laboratories, on the transplantation of various types of human cells, including peripheral blood lymphocytes and hematopoietic cells, into the scid. Insight into important applications is provided by the experiments on HIV infection and Epstein-Barr Virus related B cell lymphomas by Mosier et al. Other projects concern transplantation of human tumor cells and tumor infiltrating lymphocytes. Obviously, such strategies will facilitate experiments on possible interventions in tumor development and progression. On the whole, this is a landmark book. It reflects the increasing interest and awareness of the potential uses of the scid mouse experimental model. Anyone studying immunodeficiencies, lymphocyte differentiation, bone marrow transplantation and graft rejection will find this book extremely valuable. It also offers possible approaches to monitor interventions, focussing on human cells. It will be useful as a handbook for those searching for information on the scid mouse, and for those wishing to use them as a powerful experimental tool. Weizmann Institute Rehovot, Israel
of Science
AMIELA GLOBERSON
A Cook Book of Immunological Methods. Antibodies. A Laboratory Manual. By Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, IL, U.S.A., 1988. 726 pp. S5O. This manual is a close relative to the widely known book by Maniatis ef at. “Molecular Clonina”. the same place of birth (Cold Spring Harbor Laboratory), the same appearance and style and the same main goal-to present a detailed description of simple and reproducible methods. Immunolo~cal approaches are now as widely used by molecular biologists as biochemical and genetical methods. A large army of students and young scientists requires a manual which presents useful methods of production and isolation of antibodies as well as their application for detection and quantitation of antigens in an unsophisticated manner. This book fully complies with these requirements. In the introductory chapters, a clear simplistic picture of antibody structure and biosynthesis is given. An extended list of references helps the newcomer find additional details which may be needed for a deeper understanding of the techniques and theory. The main part of the manual is devoted to the description of methods of how to obtain polyclonal and monoclonal antibodies, purify and use them in various assays for finding antigens in solutions and cells. The book is not a collection of many known methods but a careful selection of more or 681
682
Book Reviews
less simple and efficient ones. A beginner could easily choose a precise protocol necessary to perform an immunological procedure, and the comments included would help avoid methodological traps and pitfalls. Many schemes, figures and tables provide the necessary pictorial representations that enhance the clarity together with the written text. Some investigators, especially “professional” immunologists may like to add a variation to a particular recipe since each laboratory usually has its own small tricks and secrets. But in general the authors describe widely accepted methods which allow one to obtain satisfactory and precise results. Some general comments. The authors do not constantly explain why a particular step is necessary to perform and an
appropriate reference cannot always be found for an answer to that in original publications. Some references in introductory chapters are outdated. It is quite understandable in every large publication to find inaccuracies and even errors, However, it was not so easy to spot them in the Manual. On page 630 (point 3) “pepsin” is printed instead “papain”. It is not very clear what classes of antibodies are mentioned in Table 15.1 @. 617). The recommendation to use elastase instead of pepsin (p. 631) for digestion of mouse IgG,, is questionable. Department of Chemical Immunology, Weizmann Institute of Science, Israel
ROALDNEZLIN
1991
BOOK REVIEWS
The Scid Mouse, Characterization and Potential Uses. Edited bv M. J. Bosma. R. A. Phillins and W. Schuler. Current T&s in Microbklogy and inukology, Vol. 152. Springer, Berlin, 1989, 263 pp. DMI32. This comprehensive book presents data on severe combined immunodeflcient (scid) mice. It provides their background and characterization, including cellular and molecular aspects. It reveals the many areas in immunology in which these mice can serve as useful experimental models, which allow studying questions that were heretofore unapproachable. The scid mice were discovered by Bosma in 1980. He was performing a routine screening of immunoglobulin allotypes in the heavy chain congenic BALB/c and C.B-17 mouse strains, when he noticed that some of the Cb-17 mice did not express serum immunoglobulin of the appropriate ahotype. Further checking revealed that the mice actually lacked all of the major immunoglobulin classes, and it became apparent that these were mutant mice. By 1987, the uniqueness and versatility of the scid mouse model was well recognized. At that time the European Molecular Biology Organi~tion (EMBO) held a workshop on the subject and this book is a collection of the papers presented thereat. The book is impressive in its scope, covering the full range of important research issues that have been investigated on these mice. Data provided by Bosma point out the developmental stages in the T and B lineages at which differentiation ceases. The mutation was found to interrupt the process of gene-rearrangement, due to a defective V(D)J recombinase system in developing lymphoid cells. It is manifested in prevention of the proper expression of antigen receptors on both B and T lymphocytes. The scid mice thus offer a special expe~mental model for studies of lymphoid development. A detailed study on hemopoiesis in the scid mouse is reported by Phillip and Fullop, showing normal development of myeloid and NK cells and absence of mature lymphoid elements. These studies led also to the observation on increased radiation sensitivity in these mice, which is of particular interest in relation to Ataxia telangiectasia in the human. Extensive analysis of B and T lymphocyte development are reported from several laboratories. Noteworthy examples are Hardy et ~1,“sexperiments illustrating sequential stages of development within the bone marrow B cell compartment by dete~ination of cell membrane phenotypes, and von Boehmer’s analysis of T lymphocyte development, which makes use of the scid along with mice transgenic for the T cell receptor (TCR), which contain productively arranged alpha and beta TCR genes, and of crosses between these mice. A considerable volume of data relates to potential uses for the scid mice. They include studies on the transplantation of lymphohemopoietic cells from normal mice to examine reconstitution of the different cellular elements in intact and irradiated recipients, on grafting of bone marrow and fetal liver cells, and of fetal primordia, for examination of lymphoid developments and studies of myeloid and lymphoid stem cells. The studies on bone marrow transplantation, focussing on mechanisms underlying graft rejection, further emphasize the advantages of these mice. A study by Bennet et al. examines extensively the histocompatibility differences
identified by the scid mice, and analyzes the cellular mechanisms involved. Since graft rejection in these mice cannot be related to either T or B cells, the cause of bone marrow rejection may be NK cells, which are functional in the scid mice. Other particularly interesting potential uses include engraftment of human cells. The idea is to devise in viuo culture conditions that will allow studies on isolated human cells. Once this is achieved, a whole new spectrum of studies will be feasible. The part of the book devoted to this area includes research from different laboratories, on the transplantation of various types of human cells, including peripheral blood lymphocytes and hematopoietic cells, into the scid. Insight into important applications is provided by the experiments on HIV infection and Epstein-Barr Virus related B cell lymphomas by Mosier et al. Other projects concern transplantation of human tumor cells and tumor infiltrating lymphocytes. Obviously, such strategies will facilitate experiments on possible interventions in tumor development and progression. On the whole, this is a landmark book. It reflects the increasing interest and awareness of the potential uses of the scid mouse experimental model. Anyone studying immunodeficiencies, lymphocyte differentiation, bone marrow transplantation and graft rejection will find this book extremely valuable. It also offers possible approaches to monitor interventions, focussing on human cells. It will be useful as a handbook for those searching for information on the scid mouse, and for those wishing to use them as a powerful experimental tool. Weizmann Institute Rehovot, Israel
of Science
AMIELA GLOBERSON
A Cook Book of Immunological Methods. Antibodies. A Laboratory Manual. By Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, IL, U.S.A., 1988. 726 pp. S5O. This manual is a close relative to the widely known book by Maniatis ef at. “Molecular Clonina”. the same place of birth (Cold Spring Harbor Laboratory), the same appearance and style and the same main goal-to present a detailed description of simple and reproducible methods. Immunolo~cal approaches are now as widely used by molecular biologists as biochemical and genetical methods. A large army of students and young scientists requires a manual which presents useful methods of production and isolation of antibodies as well as their application for detection and quantitation of antigens in an unsophisticated manner. This book fully complies with these requirements. In the introductory chapters, a clear simplistic picture of antibody structure and biosynthesis is given. An extended list of references helps the newcomer find additional details which may be needed for a deeper understanding of the techniques and theory. The main part of the manual is devoted to the description of methods of how to obtain polyclonal and monoclonal antibodies, purify and use them in various assays for finding antigens in solutions and cells. The book is not a collection of many known methods but a careful selection of more or 681
682
Book Reviews
less simple and efficient ones. A beginner could easily choose a precise protocol necessary to perform an immunological procedure, and the comments included would help avoid methodological traps and pitfalls. Many schemes, figures and tables provide the necessary pictorial representations that enhance the clarity together with the written text. Some investigators, especially “professional” immunologists may like to add a variation to a particular recipe since each laboratory usually has its own small tricks and secrets. But in general the authors describe widely accepted methods which allow one to obtain satisfactory and precise results. Some general comments. The authors do not constantly explain why a particular step is necessary to perform and an
appropriate reference cannot always be found for an answer to that in original publications. Some references in introductory chapters are outdated. It is quite understandable in every large publication to find inaccuracies and even errors, However, it was not so easy to spot them in the Manual. On page 630 (point 3) “pepsin” is printed instead “papain”. It is not very clear what classes of antibodies are mentioned in Table 15.1 @. 617). The recommendation to use elastase instead of pepsin (p. 631) for digestion of mouse IgG,, is questionable. Department of Chemical Immunology, Weizmann Institute of Science, Israel
ROALDNEZLIN