Immunology Of Nude Mice

180 Pages·1989·26.399 MB·\180

by  Miroslav Holub (Author)

Immunology
o f
Nude Mice
Author
Miroslav Holub
Head, Experimental Immunology Group
Institute for Clinical and Experimental Medicine
Prague, Czechoslovakia
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PREFACE
At the 1st International Workshop on Nude Mice held in Aarhus, Denmark in 1973, 36
papers were delivered, as judged from the proceedings published 1 year later. Of these,
92% were devoted to the breeding, biology, immunology, embryology, cytology, physi
ology, pathology, and reconstitution of the mouse itself, so to speak, of the mouse personally.
Only three papers dealt with the application of this model for in vivo culture of xenografts.
At the 5th International Workshop held in Copenhagen, Denmark in 1985, 67 papers (and
posters) dealing with nude mice were presented, as evident from the proceedings (Immune-
Deficient Animals in Biomedical Research) published 2 years later. Of these, 21% were
concerned with the nature (biology, immunology) of the mouse, 53 papers and posters
addressed the use of the mouse as a living test tube.
The shift in the proportion of contributions on basic questions about the nu mutation from
92 to 21% is representative also for the situation in biomedical laboratories and journals in
general. It looks as if the nude mutation is sufficiently known to be justly applied as a
standard and routine tool. It is not, of course; not only are there some physiological problems
that have failed to trigger enough attention, but many of the basic immunological and
endocrinological studies have been performed on poorly defined animal material and with
hardly comparable methods. The purpose of this book is to survey at least some areas of
solid knowledge and to show how many questions regarding the nude mouse itself remain
unanswered.
The scope of a single author must be, by definition, limited, but writing scientific mono
graphs in this old-fashioned way has one advantage: there is less overlapping and less
background noise than in a collection of chapters/articles by single specialists.
I am much indebted to the people at CRC Press for coping with the manuscript written
in a nonmaternal language and in conditions where technological progress has to be sup
plemented by human effort and dedication. I am also deeply grateful to the Director and
workers of the Institute for Clinical and Experimental Medicine in Prague.
Miroslav Holub
THE AUTHOR
Miroslav Holub, M.D., Ph.D., is a senior scientific worker at the Institute for Clinical
and Experimental Medicine in Prague, Czechoslovakia. He heads the experimental unit of
the Institute's Immunological Department.
Dr. Holub graduated and received the M.D. degree from the Charles IV. University,
Prague, in 1953 and worked as a clinical pathologist. In 1954, he joined the newly formed
immunological laboratory in the Biological (later Microbiological) Institute of the Czech
oslovak Academy of Sciences and obtained the scientific degree, about equivalent to Ph.D.
in 1958. He was engaged in the studies of cytological aspects of antibody formation and of
the key role of the lymphocyte.
During 1965 to 1967, he worked in the immunological laboratory of the Public Health
Research Institute of New York City and in 1968 to 1969 at the Max Planck Institute for
Immunobiology in Freiburg, W. Germany. He was introduced to the nude mouse club by
the late Dr. Berenice Kindred, and has stayed on as a faithful member for all years thereafter,
with the main interest focused on the nature of thymic dysgenesis.
In 1972, he was transferred from the Academy to the Institute for Clinical and Experimental
Medicine. He has published, so far, 130 scientific papers in immunology, one monograph
(Structure of the Immune System, Prague 1979) and chapters in the Modern Trends in
Immunology 2, London 1967 and in the Grundriss der Immunbiologie, Leipzig 1978 and
1988.
During 1968 to 1970 Dr. Holub was a member of the presidium of the Union of Czech
oslovak Scientific Workers. He is also a known writer translated in most European countries,
the U.S., India, China, and Japan. In 1985, he obtained an honorary degree from Oberlin
College, Oberlin, Ohio, and is a member of the Bavarian Academy of Arts.
TABLE OF CONTENTS
Chapter 1
Introduction: The Contradictory Mouse 1
Chapter 2
The Nude Mutation 5
Chapter 3
The Thymic Defect 13
I. Introduction 13
II. The Derivation of the Epithelial Stroma 13
III. Thymic Cysts and Brown Fat 16
IV. The Mesenchymal Component 18
V. The Essential Defect in the Nude Mouse Thymus 22
VI. The Lymphatic Rudiment and the Lymphoid Infiltraton in the
Polycystic Organ 37
VII. The Thymus of the nul + Hybrid 42
VIII. Thymic Hormones 44
References 46
Chapter 4
T Cells 53
I. T-Cell Differentiation and Function 53
II. The Specificity Pattern of T Cells 56
III. Natural Killer and Cytotoxic Cells 59
References 62
Chapter 5
B Cells and Immunoglobulins 67
I. Introduction 67
II. B-Cell Turnover and Differentiation 67
III. Antibody Formation 70
IV. Tolerance 77
V. Polyclonal Activators and Background Immunoglobulins 79
References 82
Chapter 6
Stem Cells and Connective Tissues 89
I. Bone Marrow Stem Cell Potential and Leukocyte Numbers 89
II. Connective Tissues 92
III. Mast Cells 93
References 94
Chapter 7
Phagocytic System and Native Resistance 97
I. Phagocytic Cells in Ontogeny 97
II. Phagocytic Cells in Adult Nude Mice 100
References 107
Chapter 8
Lymphatic Tissues and Cellular Immune Reactions Ill
I. Lymphatic Tissues Ill
II. Cell-Mediated Immune Reactions 122
References 125
Chapter 9
Hairlessness and Metabolic Compensations 131
I. The Skin 131
II. Metabolic Rates and Deviations 135
References 137
Chapter 10
Neuronendocrine Regulations 141
I. Endocrine Organs and Functions 141
II. Thermogenesis and Neurotransmitters 146
III. Central Nervous System and Behavior 150
References 155
Chapter 11
Nude Gene Transfers and Remutation 159
Chapter 12
Conclusions 161
Index 163
1
Chapter 1
INTRODUCTION: THE CONTRADICTORY MOUSE
In a Czech fairy tale a smart mountaineer woman is charged by a man of power to come
to see him "neither in the day nor at night, neither naked nor dressed, neither on foot nor
riding". The clever girl solves the problem by coming at dawn, with a loosely woven sack
on her body, with one shoe and one sock on her feet, sitting on a goat with her legs on the
ground.
The athymic nude mouse reminds me of that viable mountain-girl-mutant; the mouse is
not athymic, because it possesses at least one component of the normal mouse thymus; it
is not nude, but it grows hair which is invisible most of the time. And lacking the key part
of the immune defenses, it is not defenseless; on the contrary it may be better equipped to
fight certain bacterial infections, at least in the early or acute phase, than a normal mouse.
Lacking the key part of the "immune surveillance" system, it has no higher incidence of
spontaneous tumors, at least not of solid epithelial tumors, and lives in a germ-free state or
in perfect specific-pathogen-free (SPF) conditions for the same 1000 days as do normal mice
of the given background, outliving even the immune surveillance theory in its original form.
The nude mutant may have been here since 1962, or indeed since 1850, when two hairless
mice died in a zoologist's grape vase and the third one escaped. Even since 1962 or 1968,
the year the immunological career of the nude mutant started, when it occurred to one of
the nude mice pioneers to check what the nude mouse had inside, the nude mouse may have
changed considerably due to nu gene transfer to different background strains and for different
levels of backcrossing, and as a result of progressively improving housing conditions,
breeding and husbandry, with consequent elimination of some known or unknown natural
pathogens. It cannot be claimed that an SPF BALB/c(10th backcross)fiw/ww mouse of 1987
is exactly the same as such a mouse from 1972. This is very good for the mouse which
nowadays has litters from homozygous (nu/nu x nu/nu) matings in almost every breeding
center — a rarity 15 years ago; but this is not good for the results or the comparability of
immunological and physiological studies (Figure 1).
In any case, the nude mouse has been a tool for an impressive amount of work on the
nature of T cells and on the interaction of T cells with MHC class I and class II antigens
(or rather molecular complexes with processed exogenous antigen). The T-cell studies were
motivated by the "athymic state" of the model, but the "athymic state" itself was not
enough of a motive for deeper embryological investigation or manipulation to pin down the
reason for the athymic state, which is as obscure today as it was in 1968. Hence, one may
read in the discussion in a paper on nude mice that "the presence of thymus cells in the
nude mouse is undoubtedly related to the presence of a thymus during the first 12 days of
gestation...".
I wonder what would have happened to the poor nude mouse if it had first fallen into the
hands of neurophysiologists — given its cortical and oligodendroglial deficits — or of
endocrinologists — given its hypothalamic or even pineal body alterations and disturbed
circadian rhythms — or of students of phagocytosis — given its impressive phagocytic
capacities. Undoubtedly, thymic dysgenesis would have been discovered eventually, and it
would have become the immunologically dominant trait, but the thinking would then not
have been so thymus-monomanic. Not every single physiological alteration would have been
attributed primarily to the athymia, which may, at best, affect the ontogeny from the 12th
to 13th gestational day onwards, but also to something that may have gone wrong earlier,
due to the nu gene.
2 Immunology of Nude Mice
FIGURE 1. C57B1.10 LP female nude mouse, 3 months old, carrying her off
spring from a homozygous litter (nu/nu x nul nú).
As Hans Gruüneberg wrote in 1948:1
On the morphological level, gene-controlled disturbances provide a unique material for the study of epigenetic
relationships... Work of this kind has as its background the postulate of the unity of primary gene action. From
this postulate it follows logicaly that the various effects of a gene on the morphological level, however ill-assorted
they may appear at first glance, are causually connected as an epigenetic pattern... That the progress made in the
analysis of these phenomena is so far slight has two main reasons. One is that one cannot usually penetrate to this
level without breaking down a solid morphological crust which hides the more fundamental processes from view.
The other is the comparatively untractable chemistry of these big molecules...
This monograph is based on the attempt not to ascribe too many phenomena within "the
solid morphological crust' ' to athymia, but to try to penetrate back to the level o fthe possible
3
Table 1
PRINCIPAL PUBLICATIONS ON NUDE MICE
Proc. 1st Int. Workshop on Nude Mice, Rygaard, J. and Povlsen, C. O., Eds., G. Fischer, Stuttgart, 1974.
Proc. 2nd Int. Workshop on Nude Mice, Nomura, T., Ohsawa, N., Tamaoki, N., and Fujiwara, K., Eds., University
of Tokyo Press, Tokyo, 1977 and G. Fischer, Stuttgart, 1977.
Proc. 3rd Int. Workshop on Nude Mice, Reed, N. D., Ed., G. Fischer, New York, 1982.
Immune-deficient Animals, Proc. 4th Int. Workshop on Immune Deficient Animals, Sordat, B., Ed., S. Karger,
Basel, 1984.
Immune-deficient Animals in Biomedical Research, Proc. 5thlnt. Workshop on Immune Deficient Animals, Rygaard,
J., Brünner, N., Graem, N., and Spang-Thomsen, M., Eds., S. Karger, Basel, 1987.
Rygaard, J., Thymus & Self, Immunobiology of the Mouse Mutant Nude, F. A. D L. Copenhagen, 1973.
Bibliography of the Nude Mouse 1966—1976, Rygaard, J. and Povlsen, C. O. Eds., G. Fischer, Stuttgart, 1977.
Proc. Symp. on the Use of Athymic (Nude) Mice in Cancer Research, Houchens, O. P. and Ovejera, A. A., Eds.,
G. Fischer, Stuttgart, 1978.
The Nude Mouse in Experimental and Clinical Research, Vols. 1 and 2, Fogh, J. and Giovanella, B.C., Eds.,
Academic Press, New York, 1978, 1982.
Immunodeficient Animals in Cancer Research, Sparrow, S., Ed., Macmillan, London, 1980.
Thy musaplastic Nude Mice and Rats in Clinical Oncology, Bastert, G., Fortmeyer, H. P., and Schmidt-Matthiesen,
H., Eds., G. Fischer, Stuttgart, 1981.
Fortmeyer, H. P., Thymusaplastische Maus (nulnu) Thymusaplastische Ratte (rnu/rnu). Haltung, Zucht, Versuchs-
modelle, Paul Parey, Berlin, 1981.
Kindred, B., Nude mice in immunology, Prog. Allergy, 26, 137, 1979.
Kindred, B., Deficient and sufficient immune systems in the nude mouse, in Immunologic Defects in Laboratory
Animals, Vol. 1, Gerschwin, M. E. and Merchant, B. Eds., Plenum Press, New York, 1981, 215.
''primary gene action", at least by showing the multitude of alterations everywhere in the
organism which has to survive the defect in one entire organ system2.
The alterations may affect other alterations, compensations may affect the defects and
may themselves be altered by other compensations. Some of the alterations may be parallel,
not consequent upon the thymic dysgenesis. In other words, the nu gene effect may well
be operating before the first thymic epithelial cells assemble to the anlage. Absence of
evidence is not evidence of absence of these ontogenic factors.
However, in view of the available monographs, special attention and discussion will be
focused on the thymus problem proper which is a neglected orphan in all nude mouse
literature, albeit an orphan blamed or made responsible for all of the nude mice.
This leaves little space for the favorite topics of nude mouse reconstitution, which were
exhaustively surveyed by Kindred (Table 1) and no space to the applications of nude mice
in experimental oncology, to which a special symposium and linearly increasing proportions
of international workshops on nude mice and other immunodeficient animals (IWIDA) are
devoted (Table 1).
A perfect introduction to the problems of nude mouse breeding and production is provided
by Fortmeyer's monograph and by specialized chapters in the book edited by Fogh and
Giovanella and in separate IWIDA proceedings (Table 1).
REFERENCES
1. Grüneberg, H., Genes and pathological development in mammals, in Growth in Relation to Differentiation
and Morphogenesis, Symposia of the Society for Experimental Biology No. 2, Cambridge University Press,
Cambridge, 1948, 155.
2. Rygaard, J., Thymus & Self, Immunobiology of the Mouse Mutant Nude, F. A. D. L., Copenhagen, 1973.