Beruflich Dokumente
Kultur Dokumente
Editors: Parslow, Tristram G.; Stites, Daniel P.; Terr, Abba I.; Imboden,
John B.
Title: Medical Immunology, 10th Edition
Copyright 漏2001 McGraw-Hill
> Table of Contents > Section I - Basic Immunology > 1 - Fundamentals of B lood Cell
Biology
1
Fundamentals of Blood Cell Biology
Immunology is the study of the ways in which the body defends itself from
infectious agents and other foreign substances in its environment. Broadly
defined, the field encompasses many layers of defense, including physical
barriers like the skin, protective chemical substances in the blood and tissue
fluids, and the physiologic reactions of tissues to injury or infection. But by far
the most elaborate, dynamic, and effective defense strategies are carried out by
cells that have evolved specialized abilities to recognize and eliminate potentially
injurious substances. Some of these defensive cells circulate continually through
the body in search of foreign invaders; others are stationary sentinels that lie in
wait in solid tissues or at body surfaces. Because of their central roles in host
defense, these cells are the major focus of contemporary immunology and are
the principal subjects of this book.
Virtually all of the specialized defensive cells have two things in common: They
all spend at least part of their lives in the bloodstream, and they are all
ultimately derived from cells produced in the bone marrow. We therefore begin,
in this chapter, by considering the processes involved in cell formation and
maturation in the bone marrow鈥攐ne of the most prolific sites of cell replication
in the human body and one that is indispensable for health and even for
survival. Investigating these processes provides an opportunity to introduce
many of the individual cell types involved in host defense, as well as several
regulatory factors that govern their lives. We will also examine the fundamental
molecular mechanisms by which cells receive signals from their environments
and how these signals control whether cells proliferate, migrate, and carry out
specific functions, and even when they die.
HEMATOPOIESIS
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some of their daughter cells remain as HSCs, so that the pool of stem cells does
not become depleted.
The other daughters of HSCs, however, can each commit to any of several
alternative differentiation pathways that lead to the production of one or more
specific types of blood cells (Figure 1-1). A typical pathway involves several
cycles of cell division (five or more) and proceeds in stages, with cells at each
stage progressively acquiring features of one particular mature cell type while
losing the capacity to form any others. Because progression along these
pathways is coupled to cell division, the more mature forms greatly outnumber
their less differentiated precursors. As the cells differentiate, however, their
capacity for replication and self-renewal declines. Indeed, most types of
hematopoietic cells lose replicative capacity altogether by the time they are fully
mature, and so are said to be terminally differentiated. Thus, in general, the
less differentiated
P.2
cells in a given pathway are rare but replicate actively, whereas the mature cells
are more numerous but mitotically inert.
Vast numbers of mature blood cells are produced daily in the marrow, but the
rate of production of each cell type is precisely controlled and responsive to
physiologic demands. For example, production of leukocytes often increases
markedly during systemic infections, whereas red cell production can rise as a
reaction to anemia. In addition, many mature leukocytes, particularly
neutrophils, are stored in the marrow before being released into the
bloodstream. This storage pool, which normally accounts for 10鈥?0% of all
marrow cells, provides a reservoir of mature defensive cells that can be
mobilized rapidly in times of need. Thus, bone marrow hematopoiesis is precisely
controlled at several levels in order to (1) maintain an available pool of HSCs;
(2) regulate the commitment, proliferation, and differentiation of cells at all
stages of each hematopoietic pathway; and (3) modulate the activity of each
pathway in response to physiologic demands. As we shall see, much of this
regulation is achieved through physical interactions of the hematopoietic cells
with other cells and with soluble factors in the surrounding tissues.
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Ontogeny of Hematopoiesis
HSCs arise in the mesoderm of the yolk sac during the first weeks of embryonic
life (Figure 1-2). Within 2 months following conception, most HSCs have
migrated to the fetal liver, and it is here that the bulk of hematopoiesis occurs
during fetal development. Most embryonic and fetal hematopoiesis is devoted to
the production of red cells; platelet production first becomes apparent at 3
months of gestation, and leukocytes do not appear until the fifth month. Later in
gestation, HSCs begin to colonize the developing bone marrow cavities
throughout the skeleton, which contain a network of epithelial cells (called the
bone marrow stroma) that provide the necessary environment for growth and
differentiation of HSC and their progeny. By birth, virtually all of the marrow
space is occupied by developing hematopoietic cells, giving the newborn child
about the same hematopoietic capacity as his or her adult parents.
Hematopoietic activity in the long bones then declines with age, so that after
puberty it is largely confined to the axial skeleton鈥攖he pelvis, sternum, ribs,
vertebrae, and skull. If the bone marrow is injured by infection or malignancy,
however, hematopoiesis can resume in the liver and spleen of an adult to
maintain the supply of blood cells.
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HSCs are small, morphologically nondescript cells with round nuclei and scant
cytoplasm. They vary markedly in their ability to proliferate and differentiate
into mature hematopoietic cells (Figure 1-4). At any given moment, about 25%
are actively progressing through mitosis, but at varying rates鈥攁dult marrow
HSCs proliferate slowly, fetal liver HSCs do so rapidly. The entire human HSC
pool probably turns over every several months. Since the total number of HSCs
in the body normally remains constant, some portion of their daughter cells must
undergo differentiation into the various mature cell types. The factors that
control this lineage commitment 鈥渄ecision鈥?for any individual HSC are
unknown. It is clear, however, that both the self-replication of HSCs and their
ability to produce differentiated cells depend on hormonal growth factors called
cytokines. The cytokines are a diverse group of polypeptides, secreted by both
hematopoietic and nonhematopoietic cells. Many cytokines have specific effects
on the growth, differentiation, survival, or function of blood cells. There are
several different classes of cytokines (see Chapter 10); most of those known to
regulate hematopoiesis belong to subgroups called the colony-stimulating
P.5
factors (CSFs) or the interleukins. Cytokines that can affect HSCs include
stem cell growth factor (SCF), Flt3-ligand, interleukin-6 (IL-6), and interleukin-
11 (IL-11). These and other cytokines are produced by bone marrow stromal and
hematopoietic cells in response to environmental stimuli (eg, anemia or
infection) to regulate HSC proliferation and differentiation. Besides self-
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such progenitors grow readily in tissue culture, they are comparatively easy to
study experimentally, and it is from such cultures that most of our knowledge of
human progenitor cells has been gained. For example, when a mixed population
of bone marrow cells is cultured as a suspension in agar or some other semisolid
medium, in the presence of specific cytokines, specific progenitors proliferate
and differentiate to produce small multicellular colonies composed of one or
more mature cell types. Such experiments are called colony-forming assays, and
the progenitors themselves are often referred to as colony-forming units
(CFUs). Using these assays, individual cytokines have been shown to promote
the growth of specific types of progenitors: For example, a CSF called
erythropoietin (EPO) enhances production of erythrocyte colonies; interleukin-5
(IL-5) favors colonies of eosinophils; and granulocyte鈥搈onocyte colony-
stimulating factor (GM-CSF) promotes colonies containing both neutrophils and
macrophages (from either HSCs or mixed marrow populations).
Progenitors of all types together account for less than 1% of marrow cells, and
fully committed progenitors are more abundant than the multipotent forms.
Although comparatively rare, these marrow progenitors are the cells that give
rise to the various common forms of cancer known as leukemias鈥攎alignant
populations of leukocytes, usually from a particular hematopoietic lineage鈥攖hat
flood the marrow cavities and peripheral blood and that result from excessive
proliferation of the progeny from a single founder cell. Most mature cell types in
the marrow, despite their greater abundance, cannot give rise to leukemias, in
part because they have lost the ability to proliferate.
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尾1 family
尾2 family
伪X尾2 Fibrinogen.
尾3 family
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E-selectin CLA b
P-selectin PSGL-1.
P.8
The cell surface protein CD44 can exist in a variety of alternative forms that
differ in their extracellular ligand-binding regions (and hence in their binding
specificities) as a result of alternative RNA splicing. Hematopoietic progenitor
cells express the most truncated form of CD44, which binds hyaluronic acid, an
abundant glycosaminoglycan found in the ECM. Compounds that interfere with
this binding block hematopoietic cell proliferation in LTBMCs.
Multilineage
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Lineage-Restricted
Synergistic
a
Properties of most of these cytokines are considered in detail in
Chapter 10.
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Thus, a wide variety of adhesive interactions, both of the cell-cell and cell-ECM
variety, are critical for hematopoiesis. Abnormal interactions between
hematopoietic progenitors and ECM components or stromal cells can be observed
in many diseases that involve abnormal hematopoiesis. For example, the
malignant cells in certain types of leukemias, particularly those arising from
myeloid cells, often show significantly reduced adhesion to ECM proteins and
stromal cells. Conversely, stromal cells from patients with leukemia often
produce abnormal cytokines and ECM protein components.
Besides providing the appropriate adhesive and ECM molecules for progenitor
cells, bone marrow stromal cells also synthesize and express a host of cytokines
needed for hematopoietic proliferation (Table 1-2). Some of these are not only
secreted but also expressed as membrane-bound proteins that remain attached
to the surface of the stromal cell. Much evidence suggests that these membrane-
bound cytokines may have much greater biologic activity on hematopoietic
progenitors than the secreted forms, presumably because the effective
concentration of the cytokine is much higher on the stromal cell surface. In
addition, the strong adhesive contacts between the two cell types prolong the
much weaker interaction between a membrane-bound cytokine and its receptor
on the hematopoietic cell. In a similar fashion, IL-3, GM-CSF, and several other
cytokines
P.9
bind tightly to glycosaminoglycans and other components of the ECM, which
immobilizes them, increases their local concentrations, and maximizes their
availability to hematopoietic cells. Localized secretion of cytokines from stromal
cells into the ECM may have a role in delineating specific microenvironments
within the bone marrow.
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Although it was long believed that specific cytokines acted by inducing HSCs and
progenitors to differentiate along a certain pathway, the weight of evidence
currently favors another view. It now appears instead that each cell is
intrinsically predisposed toward one lineage or another (apparently choosing
among them at random) but is unable to proliferate, or even survive, unless the
cytokines appropriate to that lineage are present. In other words, cytokines do
not direct cells into a particular pathway but instead act as lineage-specific
growth and survival factors. Thus, when progenitor cells are deprived of an
essential cytokine, they not only cease growing but often die鈥攁ctively
committing suicide through a process called apoptosis (see later discussion). On
the other hand, progenitors that have been genetically manipulated so that they
cannot undergo apoptosis continue growing and differentiating along a particular
lineage even when the cytokine is withdrawn, implying that the differentiative
fate of each cell is intrinsically programmed. The concept that lineage
commitment is a stochastic (ie, random) process and that one major function of
cytokines is to promote survival, rather than induce differentiation, explains why
so many cytokines seem to use so few signal transduction pathways, as will be
described in the following section.
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Figure 1-6. Cytokine receptor families. The receptor tyrosine kinase family
is characterized by the presence of tyrosine kinase domains (shaded boxes)
within the intracellular portion of the receptor protein. Hematopoietin
receptors lack tyrosine kinase domains, but their extracellular regions share
conserved cysteine residues (thick lines) and the sequence motif
TrpSerXTrpSer (narrow lines), where X is any amino acid. Hematopoietin
receptors can be composed of one, two, or three polypeptide chains.
Examples of each receptor type are listed at bottom of the figure.
Abbreviations: CSF = colony-stimulating factor; SCF = stem cell factor; IL =
interleukin; GM-CSFR = granulocyte-macrophage colony-stimulating factor
R.
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The sequences of events that occur following PTK activation and that lead to
specific cellular responses vary among different cytokines, receptors, and cell
P.11
types. Perhaps the best characterized signaling pathways are those that affect
cell proliferation or modulate the transcription of particular genes. Following the
onset of PTK activity, many such signals are commonly transmitted to the
nucleus through three primary pathways, called the Ras-dependent pathway, the
Jak-Stat pathway, and the nuclear factor 魏B(NF-魏B) pathway (Figure 1-7).
The Ras pathway is actually much more complicated than the foregoing
explanation would suggest. For example, at least three different forms of MAPK
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P.13
and result in profound deficits in hematopoiesis and in cellular defense functions.
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with a variety of adapter proteins (including TRAD, TRAF, and RIP). This
complex activates a pathway leading to phosphorylation and degradation of
I-魏B, allowing NF-魏B to migrate into the nucleus. The NF-魏B pathway is
also activated by many other stimuli, all of which function similarly through
I-魏B.
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Figure 1-8. The cell cycle. This simplified depiction illustrates a few of the
molecular factors that influence the G 1 /S transition. Abbreviations: CDK =
cyclin-dependent kinase; CDKI = CDK inhibitor; sharp arrowheads =
activation; blunt arrowheads = inhibition.
Cell cycle regulation takes place mainly at the transition points, or boundaries,
between the different phases. Each of these boundaries serves as a checkpoint:
Specific requirements (eg, completion of DNA synthesis or DNA repair) must be
met before a cell can move through the checkpoint from one phase to the next.
In mammals, most variation in cell cycle timing is due to variation in the length
of G 1 , and the G 1 /S checkpoint is tightly regulated. Three main classes of
proteins are directly involved in checkpoint regulation: (1) the cyclins, whose
levels rise and fall in specific phases of the cycle; (2) the cyclin-dependent
kinases (CDKs),
P.14
a class of serine鈥搕hreonine kinases that regulate cyclin activity by
phosphorylation; and (3) the CDK inhibitors (CDKIs), which inactivate the CDKs.
There are at least seven different cyclins and CDKs, each operative at different
phases of the cycle. The intracellular levels of specific cyclins are in part
responsive to external stimuli: For example, CSF-1 acts on cells of the monocyte
lineage to increase expression of cyclins required for progression through the
G 1 /S checkpoint. The expression levels of particular cyclins can be used to
determine which stage of the cell cycle a population of cells has reached. As
cyclin levels increase during a given phase, they form a complex with the
corresponding CDKs; this activates the CDKs, which then phosphorylate other
substrates that allow checkpoint progression. In hematopoietic cells, for
example, one of the predominant species that accumulates during G 1 is the
cyclinD/ CDK6 complex, whose primary substrate is the nuclear retinoblastoma
(Rb) protein. Rb ordinarily exists as a complex with transcription factors of the
E2F family. When the cyclinD/CDK6 complex hyperphosphorylates Rb, the E2F
proteins dissociate from it and instead bind and activate specific genes, such as
those encoding DNA polymerase and thymidine synthetase, which are required
for DNA synthesis and, hence, for entry into the S phase.
Other signals can inhibit cell cycle progression by inducing CDKIs. The CDKIs are
small molecules, numbered according to their molecular weights, which bind and
inhibit specific cyclin/CDK complexes. In hematopoietic cells, for example, the
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high. These tidy, inconspicuous suicides are very different from the uncontrolled,
accidental form of cell death called necrosis, in which the dying cell often swells
and then bursts, producing further damage by spewing its contents onto its
neighbors.
The biochemical processes that occur during the final (鈥渆xecution鈥? phase of
apoptosis are thought to be similar in all cell types. Perhaps most important is
the activation of cytoplasmic proteases called
P.15
caspases, which are distinguished from other proteases by having an essential
cysteine in their active sites and by their preference for cleaving target proteins
at particular aspartate residues. Caspases are normally expressed in the
cytoplasm as larger, inactive precursors, called procaspases, but become
activated when they are cleaved proteolytically, usually by other caspases. This
allows the caspases to activate one another in a self-amplifying, proteolytic
cascade. More than ten different human caspases have been identified, at least
five of which become activated during apoptosis. Some (including caspases-8
and -9) have large prodomains that can interact with specific regulatory
proteins; these are often the first caspases to become active during an apoptotic
response, whereas those with shorter prodomains (such as caspases-3, -6, and -
7) generally become active later in the cascade.
In addition to cleaving one another, the activated caspases directly attack other
cellular proteins, producing many of the hallmarks of apoptotic death. For
example, caspases break down structural proteins of the nuclear matrix and
cytoskeleton, leading to collapse of the nucleus and cytoplasm. Proteins required
for cell鈥揷ell adhesion are also broken down, causing the cell to detach from its
neighbors, round up, and become easier to engulf. Caspases proteolytically
activate a cellular endonuclease that then attacks and degrades chromosomal
DNA, and they simultaneously destroy DNA repair enzymes that might otherwise
limit the damage. Numerous signaling molecules, cell cycle regulators, and
transcription factors are also degraded, crippling the cell's vital functions. The
importance of caspases as cellular executioners is confirmed by the finding that
drugs that inhibit these enzymes can completely prevent apoptosis in laboratory
studies. Perhaps for this reason, human cells encode at least four related
proteins, called inhibitors of apoptosis (IAPs), that can directly bind and inhibit
caspases; their physiologic role is not yet clear. The same strategy is used by
some poxviruses and herpesviruses, which encode caspase-inhibitor proteins that
prevent infected cells from committing apoptosis.
The procaspases needed for suicide are always ready and can be activated by
various signals from inside or outside the cell. One important receptor for death
signals is a surface protein called Fas (or CD95). Many cells express Fas when
they are suicide-prone because it allows them to be killed by other cells
expressing a surface protein called Fas ligand (FasL). Contact with a FasL-
bearing cell causes clustering of Fas receptors, whose cytoplasmic tails then
associate with specific adapter proteins in the cytoplasm (Figure 1-9). These
adapters then bind and activate procaspase-8 which, in turn, triggers the rest of
the caspase cascade and leads quickly to apoptosis. (In some cases, stressed
cells express both Fas and FasL on their surfaces, and so trigger their own
death!) Binding of TNF伪 by the TNF receptor can likewise lead to caspase
activation, as well as activating the NF-魏B pathway, as previously described. An
even more direct approach can be used by specialized lymphocytes to kill virally
infected cells. These lymphocytes bind the target cell, create minute openings in
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its surface, and secrete into its cytoplasm a protease called granzyme B, which
cleaves and activates multiple caspases, causing apoptosis (see Chapter 9).
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Bcl2 Bax
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BclX L BclX S
Mcl-1 Bak
A-1 Bik
p35 (Baculovirus)
Ced-9 (Nematode)
a
Each of these proteins is expressed by mammalian cells unless
otherwise indicated. BclX L and BclX S are alternatively spliced products
from a single gene.
P.16
The mechanisms by which Bcl2 and related proteins regulate apoptosis are not
fully known. Their structures resemble those of certain bacterial pore-forming
proteins, and Bax and Bcl2 have each been shown to form ion channels with
somewhat different properties in artificial membranes. Bax and Bid normally
reside in the cytosol but move to mitochondria after apoptotic stimulation, when
they trigger the release of cytochrome c into the cytoplasm. By contrast, Bcl2
antagonizes this release, and BclX L may interfere with cytochrome c-mediated
activation of
P.17
caspase-9. It also appears that some cytokines regulate the expression of Bcl2
family members. For example, withdrawing a particular cytokine may lead to
reduced Bcl2 expression in some hematopoietic progenitors, leaving these cells
vulnerable to the unopposed effects of Bax. Certain viruses also manipulate
expression of these proteins: For example, the Epstein-Barr virus not only
encodes a Bcl2-like protein of its own, but also specifically activates expression
of cellular Bcl2, thereby preventing the infected cell from committing apoptosis.
CONCLUSION
The regulation of hematopoiesis reflects the individual and combined effects of
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1988;10:271.
Obaya AJ et al: Mysterious liaisons: The relationship between c-Myc and the
cell cycle. Oncogene 1999;18: 2934.
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APOPTOSIS
Wickremasinghe RG, Hoffbrand AV: Biochemical and genetic control of
apoptosis: Relevance to normal hematopoiesis and hematological
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Lundberg AS, Weinberg RA: Control of the cell cycle and apoptosis. Eur J
Cancer 1999;35:531.
Chao DT, Korsmeyer SJ: Bcl-2 family: Regulators of cell death. Ann Rev
Immunol 1998;16:395.
*The term myeloid means 鈥渙f the bone marrow.鈥?As a group, cells of the
myeloid lineage are the most abundant cells in the marrow.
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