Immune escape of tumors: apoptosis resistance
and tumor counterattack
Frederik H. Igney and Peter H. Krammer
Tumor Immunology Program, German Cancer Research Center (DKFZ), Heidelberg, Germany
Abstract: Interactions between the immune sys-
tem and malignant cells play an important role in
tumorigenesis. Failure of the immune system to
detect and reject transformed cells may lead to
cancer development. Tumors use multiple mecha-
nisms to escape from immune-mediated rejection.
Many of these mechanisms are now known on a
cellular and molecular level. Despite this knowl-
edge, cancer immunotherapy is still not an estab-
lished treatment in the clinic. This review discusses
the immune escape mechanisms used by tumors
with an emphasis on mechanisms related to
apoptosis. J. Leukoc. Biol. 71: 907920; 2002.
Key Words: CD95L cytotoxic T cells CD95 death receptor
immunosuppression
TUMORS AND THE IMMUNE SYSTEM
The immune system has evolved in order to detect and elimi-
nate pathogens that may do harm to the organism. Moreover, it
serves as a watchdog against transformed cells that may lead to
cancer [1, 2]. The key cells of the immune system for tumor
surveillance are T cells, which are part of the adaptive immune
response. After recognition of an antigen on a tumor cell via the
T cell receptor (TCR), activated CD8 T cells can kill the
tumor target cell and thus are called cytotoxic T cells (CTL).
One subset of CD4 T cells, T helper cell type 1 (Th1),
provides help for the activation of CD8 T cells. The other
CD4 subset, Th2 cells, stimulates a humoral immune re-
sponse and suppresses the development of a Th1 response.
CD4 T cells can also display cytotoxic activity in some
situations. CD8 and CD4 T cells recognize antigens pre-
sented as peptides by major histocompatibility complex (MHC)
class I or class II molecules, respectively.
Numerous tumor antigens have been identified that can be
recognized by T cells [35]. Some of these antigens are ex-
pressed exclusively by tumors and thus are called tumor-
specific antigens. These antigens arise from mutations or trans-
locations of normal cellular genes (e.g., catenin, cdk4, ras).
The mutations may be involved directly in carcinogenesis.
Another group of antigens are the tumor-associated antigens
that are not only expressed by tumor cells, but are also ex-
pressed by other cells of the body. Cancer-testes antigens are
expressed on a variety of epithelial tumors as well as on testis
and placental tissue (e.g., MAGE, BAGE, GAGE). Overex-
pressed nonmutated proteins (e.g., p53, Her2/neu) may also
serve as tumor antigens for T cells. In addition, tumor-infil-
trating lymphocytes have been identified that are reactive
against differentiation antigens present on normal melanocytes
as well as melanomas (e.g., MART-1/Melan-A, tyrosinase,
gp100). Moreover, antigens from tumorigenic viruses are pre-
sented on tumor cells. The expression of tumor antigens may be
heterogeneous within a tumor, and a single patient can develop
immune reactions to multiple antigens [6, 7].
Two different models for the immune response to tumors
have been proposed: the concept of immunosurveillance and
the danger model. According to the immunosurveillance hy-
pothesis, tumors expressing antigens are regarded as nonself
by the immune system, and a major function of the immune
system is to survey the body for the development of malignancy
and to eliminate tumor cells as they arise [8]. To detect
danger, the immune system uses professional antigen-pre-
senting cells (APC) as sentinels of tissue damage. In the
presence of danger signals, APCsuch as dendritic cells,
activated macrophages, and B cellsstimulate the T cell re-
sponse. The danger model proposes that cancer cells do not
appear dangerous to the immune system, so that the response
of T cells to tumors is not initiated [9].
Natural killer (NK) cells of the innate immune system also
play an important role in immune surveillance of tumors [1].
NK cells kill MHC class I-deficient cellsa phenomenon that
is part of the missing self hypothesis [10, 11]. The activity of
NK cells is controlled by a balance of positive and negative
signals. Engagement of inhibitory receptors by MHC class I
molecules blocks activation signals. Two families of inhibitory
receptors have been identified in humans: the immunoglobu-
lin-like killer cell inhibitory receptors and the lectin-like
CD94-NKG2 receptors. Stimulatory receptors comprise recep-
tors (e.g., CD16, CD94-NKG2C, natural cytotoxicity receptors)
that are supposed to bind to constitutively expressed ligands
[12] and NKG2d receptors, which bind to molecules that are
induced by cellular stress [1315]. Ligands for NKG2d recep-
tors are the MHC class I chain-related (MIC) glycoproteins
MICA and MICB in humans and the minor histocompatibility
antigen H60 and the retionic acid early inducible (Rae-1)
family in mice.
Correspondence: Prof. Dr. Peter H. Krammer, Tumor Immunology Program,
German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, D-69120
Heidelberg, Germany. E-mail: P.Krammer@dkfz-heidelberg.de
Received February 3, 2002; revised March 18, 2002; accepted March 20,
2002.
Journal of Leukocyte Biology Volume 71, June 2002 907
 Additional cells of the innate immune system involved in
immunity against tumors are macrophages and neutrophils
[16 19]. They can reject tumors by direct killing of the tumor
cells, by destruction of tumor vessels and matrix, and by
inhibition of angiogenesis. Moreover, they display tumor anti-
gens and can stimulate other immune cells such as CTL, NK
cells, or APC. In contrast, inflammatory cells may also con-
tribute to tumor progression by production of tumor growth
factors and stimulation of angiogenesis [20]. Macrophages and
neutrophils are recruited to the tumor site by expression of
adhesion molecules on endothelial cells and by chemotactic
proteins.
TUMOR IMMUNE ESCAPE MECHANISMS
Despite presentation of antigens by malignant cells and the
presence of immune cells that could potentially react against
these cells, in many cases the immune system does not get
activated but ignores the tumor [2, 21]. According to the
danger model, APC may not get activated in this situation due
to of a lack of danger signals [9]. Other factors may also
contribute to immunological ignorance. The immune system
ignores tumor cells, which fail to migrate to lymph nodes and
fail to activate T cells directly [22, 23]. In addition, tumors
growing in immune privileged sites such as the brain or the eye
are not surveilled by the immune system [24]. Down-regulation
of adhesion molecules in malignant tissue may inhibit immune
infiltration and thus may also contribute to immunological
ignorance [2527]. The tumor stroma has also been shown to
play a critical role [23, 28]. It may serve as a physical barrier
between the tumor and immune cells.
Growth of antigenic tumors in the presence of potent im-
mune cells cannot be explained by immunological ignorance
alone. A major goal of cancer immunotherapy is to generate an
anti-tumor immune response, e.g., by vaccination with cancer
cells fused with APC or by transfer of anti-tumor T cells.
TABLE 1. Tumor Immune Escape Mechanisms
Strategy
Ignorance
Impaired antigen presentation
Expression of immunosuppressive factors and molecules
Tolerance induction
Apoptosis resistance
Counterattack (?)
908 Journal of Leukocyte Biology Volume 71, June 2002
However, many of these approaches do not efficiently stimulate
immunity, or the tumors continue to grow despite the presence
of an immune reaction [1, 3, 29]. Multiple mechanisms have
been identified that tumors use to escape from rejection (Table
1) [30 32]. It is likely that malignant escape variants are
selected by the immune system.
One strategy to escape from the adaptive immune response
is by impaired antigen presentation. In general, defects in
antigen presentation are more pronounced in metastatic lesions
than in the primary tumor. In some tumors, expression of the
tumor antigen is down-regulated, leading to enhanced tumor
incidence and metastasis [3335]. Moreover, mutations of the
antigen can result in escape from the initial response and
contribute to the heterogeneity of tumor lesions [36]. The
heterogeneous expression of multiple antigens may hinder the
establishment of an efficient specific immune response. In
addition, reduced MHC-I expression prevents recognition of
tumor cells by the immune system [37, 38]. Tumors frequently
have a heterogeneous pattern of MHC-I expression. Total loss
of MHC-I is mainly caused by mutations of the 2 microglobu-
lin subunit [39]. Gene deletions or rearrangements, point mu-
tations, and defects in transcriptional regulation lead to selec-
tive loss of an MHC haplotype, locus, or allele [27, 35, 40 43].
Total MHC-I down-regulation may target the tumor for NK cell
attack. Therefore, the tumor needs further mechanisms to resist
NK cell-mediated lysis, such as the expression of MHC-I
surrogates [44, 45]. Reduced presentation of the tumor antigen
is also achieved by defects in the antigen processing machin-
ery. Down-regulation of the proteasome subunits LMP2 (low
molecular mass polypeptide 2) and LMP7 changes the spec-
trum of peptides presented by MHC molecules [46 48]. Two
proteins involved in loading antigenic peptides onto MHC-I
molecules, TAP (transporter associated with antigen process-
ing) and tapasin, are also frequently mutated or down-regulated
in tumor cells [37, 38, 46 50]. TAP deficiency results in loss
of MHC-I expression and an increase in tumorigenesis. More-
over, some proteins of tumorigenic viruses are not efficiently
Mechanism
Lack of danger signals
Lack of tumor antigens in lymphoid organs
Growth in immune privileged sites
Lack of adhesion molecules
Physical barrier by stroma
Mutation or downregulation of tumor antigens
Mutation or downregulation of MHC genes
Defects in antigen processing (e.g., TAP, LMP deficiency)
Cytokines (TGF-, IL-10, VEGF, etc.)
Prostaglandines
RCAS1
Anergy induction (lack of costimulatory molecules)
Immune deviation
Regulatory T cells
T cell deletion
Expression of anti-apoptotic molecules
Downregulation and mutation of pro-apoptotic molecules
CD95L expression
Expression of other death-receptor ligands
http://www.jleukbio.org
presented because they interfere with their proteosomal prote-
olysis, e.g., by mutation of the clevage site [51].
Another strategy that tumors use to escape from immune-
mediated rejection is the expression of immunosuppressive
factors [18, 52]. These factors may be expressed by the malig-
nant cells themselves or by noncancerous cells present at the
tumor site, such as immune, epithelial, or stromal cells. The
most prominent of these factors is transforming growth factor
(TGF-) [5358]. TGF- is a cytokine that affects prolifera-
tion, activation, and differentiation of cells of innate and adap-
tive immunity and thus inhibits the anti-tumor immune re-
sponse. Moreover, vascular endothelial growth factor (VEGF) is
produced by many tumor cells [59]. Besides its angiogenic
properties, it inhibits the differentiation of progenitors into
dendritic cells. Further immunosuppressive factors expressed
by malignant cells are prostaglandins [60 62], interleukin
(IL)-10 [63], macrophage-colony stimulating factor [64, 65],
and soluble tumor gangliosides [66]. The membrane protein
RCAS1 (receptor-binding cancer antigen expressed on SiSo
cells) inhibits proliferation and induces apoptosis in T cells in
vitro, suggesting a role of this molecule in immune evasion of
tumors [67].
The lack of a T cell response against tumor-associated
antigens that are also expressed by other cells of the body or
during development may be explained by tolerance [30, 68]. In
healthy organisms, self-reactive T cells are tolerized mainly by
deletion in the thymus, a process known as central tolerance.
The mechanisms of peripheral tolerance induction prevent
autoimmunity by tolerizing T cells that have escaped the
process of central tolerance. Peripheral tolerance induction is
a complex multistep process [69, 70], but in principle, four
major mechanisms can be distinguished. One mechanism is
the induction of anergy. T cell activation requires two signals,
binding of a peptide-MHC complex to the TCR and binding of
costimulatory molecules (e.g., B7) to their ligands (CD28) on
the T cell surface [71, 72]. If a T cell binds via its TCR to a
peptide-MHC complex on the target cell without sufficient
costimulation, the T cell is rendered anergic and does not
become activated when restimulated with antigen. Many tumor
cells do not express costimulatory molecules and thus may
induce anergy in anti-tumor lymphocytes [27, 7375]. Another
process of tolerance induction that tumors exploit is immune
deviation. In this process, the immune response is driven
toward a Th2 humoral response away from a Th1 response
required for efficient tumor rejection by cytotoxic T cells. The
mechanism of immune deviation is not exactly understood, but
it may depend on secretion of TGF- and IL-10 [76] or on the
presentation of the tumor antigen by B cells to CD4 Th cells
[77]. Tumors can also induce the generation of regulatory T
cells [78]. Although the molecular mechanism is not clear, a
subset of CD4 T cells seems to suppress the response of
cytotoxic T cells against tumors in some settings [79, 80]. A
further mechanism to establish peripheral tolerance to self-
antigens is T cell deletion. Repetitive stimulation of T cells
with the antigen induces apoptosis, a process referred to as
activation-induced cell death (AICD). Thus, administration of
antigens, such as superantigens, peptides, or allogeneic cells,
or direct restimulation of the TCR by anti-CD3 antibodies has
been shown to induce tolerance by T cell deletion [81 84].
This process is mainly mediated via the CD95/CD95L death
system [85 88]. Costimulation via CD28 can rescue T cells
from AICD [89], implicating another important role for expres-
sion of B7 on tumor cells. Tumors have been shown to induce
tolerance by deleting anti-tumor T cells [90, 91]. AICD, as a
result of chronic stimulation with the tumor antigen may con-
tribute to immune escape, yet the significance of this process
has not directly been shown.
Two further strategies used by tumors to evade rejection by
the immune system are related to apoptosis. First, malignant
cells have changes in the expression of molecules involved in
apoptosis signaling, resulting in resistance of the tumor to the
killing mechanisms of the immune system. Second, tumors may
adopt a killing mechanism from cytotoxic immune cells to
delete the attacking anti-tumor lymphocytes, a concept called
tumor counterattack. These apoptosis-related immune-es-
cape mechanisms will be discussed in detail below.
KILLING MECHANISMS OF THE IMMUNE
SYSTEM
T cells and NK cells use two major mechanisms to kill tumor
cells: the death receptor pathway and the granule exocytosis
pathway [92]. In the death receptor pathway, the lymphocyte
displays the death ligand CD95L on the cell surface, triggering
apoptosis via the death receptor CD95 on the target cell
[9395]. Moreover, for immune surveillance of tumors and
metastases, NK cells also use the death ligand TRAIL [tumor
necrosis factor (TNF)-related apoptosis-inducing ligand],
which triggers apoptosis via the death receptors TRAIL-R1 or
TRAIL-R2 [96 98]. Death receptors are members of the TNF
receptor superfamily and comprise a subfamily characterized
by an intracellular domain, the death domain [99, 100]. The
so-called decoy receptors are closely related to the death
receptors and lack a functional death domain [101, 102]. Death
receptors are activated by their natural ligands, co-evolved as
a death ligand family, called the TNF family (Fig. 1). When
the respective ligand binds to the death receptor, the death
domains attract intracellular adaptor proteins, which, in turn,
recruit the proform of the initiator caspase 8. (Caspase-10
may also be an initiator caspase, but this is still controversial)
[103107]. The resulting protein complex is called death-
inducing signaling complex (DISC). At the DISC, procaspase-8
is cleaved autocatalytically and yields the active initiator
caspase-8. In some cells, so-called type I cells, the amount of
active caspase-8 formed at the DISC is sufficient to initiate
apoptosis directly. In type II cells, the amount is too small, and
mitochondria are used as amplifiers of the apoptotic signal
[108]. Activation of mitochondria is mediated by the Bcl-2
homology (BH)3-only Bcl-2 family member Bid, which is
cleaved by active caspase-8 and translocates to the mitochon-
dria. After activation, mitochondria release cytochrome c, ap-
optosis-inducing factor, and other apoptogenic factors from the
intermembrane space to the cytosol [109, 110]. Concomitantly,
the mitochondrial transmembrane potential m drops. In the
cytosol, cytochrome c forms a complex with Apaf-1, adenosine
5-triphosphate, and procaspase-9. This complex is called
Igney and Krammer Apoptosis and tumor immune escape 909
Fig. 1. Apoptosis signaling via death recep-
tors. Binding of death ligands (shown here for
CD95L) leads to formation of the DISC. In the
DISC, the initiator caspase-8 is activated.
(Whether caspase-10 is a true initiator caspase
is still controversial.) Cleavage of Bid by
caspase-8 activates the mitochondrial pathway
and can be used to amplify the apoptotic signal.
Activation of mitochondria leads to cytochrome
c release into the cytosol and formation of the
apoptosome. At the apoptosome, caspase-9 is
activated. Caspase-8 and -9 activate execu-
tioner caspases, which in turn cleave the death
substrates, eventually resulting in apoptosis.
Apoptosis can be inhibited on different levels
by antiapoptotic proteins (shown in red).
apoptosome. Within the apoptosome, the initiator caspase-9 is
activated.
Activated initiator caspases cleave and activate execu-
tioner caspases, mainly caspase-3, -6, and -7 [111]. The
active executioner caspases then cleave each other, and thus
start an amplifying proteolytic cascade of caspase activation.
The active executioner caspases cleave cellular substrates, the
death substrates, leading to the biochemical and morpholog-
ical changes characteristic of apoptosis [111].
Various proteins regulate the apoptotic pathway at different
levels. FLIPs (FLICE-inhibitory proteins) interfere with the
initiation of apoptosis directly at the level of death receptors
[112]. Two splice varians have been identified in human cells,
a long form (cFLIPL) and a short form (cFLIPS). They share
structural homology with procaspase-8, but lack its catalytic
site. This structure enables them to bind to the DISC, thereby
inhibiting the processing and activation of the initiator
caspases.
A major class of regulatory proteins are members of the
Bcl-2 family that regulate apoptosis at the mitochondrial level
[109, 110]. According to their function, Bcl-2 family members
can be divided into antiapoptotic and proapoptotic proteins.
Bcl-2 family proteins influence the permeability of the mito-
chondrial membrane; however, the biochemical mechanism of
their function is not entirely clear.
A third class of regulatory proteins are the IAPs (inhibitor of
apoptosis proteins), which bind to and inhibit caspases [113].
They may also function as ubiquitin ligases, promoting the
degradation of the caspases bound. However, not all IAPs have
been shown to suppress apoptosis, and some of them may also
have functions other than caspase inhibition. IAPs themselves
are inhibited by a protein, Smac/DIABLO [114, 115], which is
910 Journal of Leukocyte Biology Volume 71, June 2002
released from mitochondria along with cytochrome c during
apoptosis and promotes caspase activation by binding to IAPs.
In the calcium-dependent granule exocytosis pathway, lym-
phocytes secrete perforin and granzymes from cytotoxic gran-
ules toward the target cell. In the presence of calcium, perforin
polymerizes and initiates as yet ill-defined changes in the
target cell membrane, which allow granzymes to pass into the
cell [116 118]. Granzymes are neutral serine proteases that
can activate caspases in the target cell [119 121]. In addition,
granzyme B may directly cleave the Bcl-2 family member Bid,
initiating the mitochondrial death pathway [122]. Perforin-
deficient mice are very susceptible to some carcinogens and
oncogenic viruses [123] and develop spontaneous lymphomas
with age [124]. Although it is clear that granzymes are indis-
pensable effector molecules in a granule exocytosis-mediated
host defense against viral pathogens [125], their contribution to
tumor rejection remains controversial [126]. Thus, mice defi-
cient for granzymes A and B are capable of rejecting tumors in
an efficient way.
Macrophages and neutrophils use totally different killing
mechanisms. In principle, they use four kinds of cytotoxic
molecules [127]. The first effector mechanism is the oxidative
burst consisting of the release of reactive oxygen species
(superoxide anion, hydrogen peroxide, and derivatives) pro-
duced by the phagocytic reduced nicotinamide adenine dinu-
cleotide phosphate oxidase (NADPH) [128 130]. Reaction of
peroxide via the myeloperoxidase pathway can yield hypochlo-
rous acid and chloramines. A further cytotoxicity mechanism is
the production of nitric oxide (NO) by the inducible NO syn-
thase [129, 131]. The toxicity of NO is enhanced greatly by
reacting with superoxide to form peroxynitrite. The molecular
targets of reactive oxygen species, NO, and derivatives inside
http://www.jleukbio.org
the target cells are not fully defined yet, but may include
enzymes essential for cellular survival. Phagocytes also release
antimicrobial peptides, defensins and cathelicidins, which
have affinity for bacterial and eukaryotic membranes and may
lyse cells by disrupting the phospholipid bilayer [132]. Other
mechanisms include the production and release of a variety of
proteases including elastase, proteinase 3, and metallopro-
teases [127]. These proteases degrade extracellular matrix
components and other proteins.
RESISTANCE TO APOPTOSIS AND IMMUNE
ESCAPE
Resistance of tumor cells to the effector mechanisms of the
immnue system leads not only to escape of the tumors from
immunosurveillance, but may also dramatically influence the
efficacy of immunotherapy. In an immune-competent host,
tumor cells are selected for resistance against the effector
mechanisms of the immune system, a concept known as im-
munoselection or immunoediting. This concept has been con-
firmed by several experiments. Tumor cells derived from T
cell-deficient RAG2/ mice grew progressively in RAG2/ Bcl-2 family members Bcl-xL and Mcl-1 are up-regulated in
mice, but were rejected in wild-type mice [133]. Tumors de-
rived from wild-type mice grew at a similar rate in both mouse
strains. Moreover, a significant proportion of aging mice with a
mutated CD95L (gld mice) develop B cell malignancies [134].
These B cell lymphomas grew and metastasized in immmuno-
deficient mice but were rejected by immunocompetent mice.
Furthermore, neutraliziation of TRAIL promoted tumor devel-
opment in mice inoculated with a chemical carcinogen [98].
Tumor cells derived from these animals were sensitive to
TRAIL-mediated apoptosis, whereas those from control mice
were not. Similarly, spontaneous lymphomas from perforin
knockout mice were rejected by T cells when transplanted into
wild-type animals, but grew in perforin knockout mice [124].
Although many mechanisms of tumor resistance to apoptosis
have been identified (Fig. 1), only a few of them have directly
been shown to be involved in immune escape. One strategy
tumors use to acquire apoptosis resistance is the overexpres-
sion of antiapoptotic molecules. The antiapoptotic proteins
FLIPL,S interfere with apoptosis induction at the level of death
receptors, but do not prevent apoptosis by perforin/granzyme
[112, 135, 136]. Nevertheless, overexpression of FLIP medi-
ates the immune escape of tumors in mouse models. Tumors
with high expression levels of FLIPL were shown to escape
from T cell-mediated immunity in vivo despite the presence of
the perforin/granzyme pathway [137]. In vivo tumor cells were
selected for elevated FLIPL levels. FLIPL overepxression also
prevents rejection of tumors by perforin-deficient NK cells
[138]. Human melanomas were shown to express high levels of
FLIP [135, 139, 140]. Moreover, in Epstein-Barr virus-positive
Burkitts lymphoma cell lines, an increased ratio of FLIP to
caspase-8 correlated with resistance to CD95-mediated apo-
ptosis [141]. Viral analogs of FLIP, viral FLIPs (v-FLIPs), are
encoded by some tumorigenic viruses, such as Kaposi sar-
coma-associated herpesvirus (KSHV) [142144]. KSHV-FLIP
promotes tumor establishment and growth in immunocompe-
tent mice by prevention of death receptor-induced apoptosis
triggered by T cells [145]. Therefore, v-FLIPs may contribute
to immune escape of v-FLIP-encoding viruses.
Further antiapoptotic proteins are involved in apoptosis
resistance of tumors, although the significance for immune
escape is less clear. Enhanced Bcl-2 expression is found in
follicular B-cell lymphoma (Bcl) with the chromosomal trans-
location t(14;18) that couples the Bcl-2 gene to the immuno-
globulin heavy chain locus [146 149]. In cooperation with the
oncogenes c-Myc or promyelocytic leukemia retinoic acid re-
ceptor , Bcl-2 contributes to tumorigenesis [150 153]. In
some studies, high Bcl-2 expression correlates with the grade
of malignancy of human tumors [154 156]. Moreover, it has
been shown in in vitro and in vivo models that Bcl-2 expression
confers resistance to CD95L and other apoptosis stimuli [154,
157159]. In some types of tumors, high Bcl-2 expression
appears to be predictive of a poor disease-free survival [155,
156, 160]. The tumor-associated viruses Epstein-Barr virus
and human KSHV encode proteins that are homologs of Bcl-2.
Both proteins, BHRF1 or KSbcl-2 (vBcl-2), respectively, have
an antiapoptotic function and enhance survival of the infected
cells [161164]. Thus, they may contribute to apoptosis resis-
tance of virus-induced tumors. In addition, the antiapoptotic
tumors and can confer resistance to multiple apoptosis-induc-
ing pathways [159, 165168].
The IAP family member Survivin is expressed in a highly
tumor-specific manner [169]. It is found in the vast majority of
human tumors, but not in normal adult tissues [170]. In neu-
roblastomas, expression correlates with a more aggressive and
unfavorable disease [171]. Besides its antiapoptotic activity,
Survivin also has an apparent role in the cell cycle [169]. In
transgenic mice expressing Survivin in the skin, its antiapop-
totic function was more prominent than its role in cell division;
Survivin, however, did not affect CD95-induced cell death
[172]. Expression of a mutant of Survivin that could not be
phosphorylated induces cytochrome c release and cell death.
In xenograft tumor models, this mutant suppressed tumor
growth and reduced intraperitoneal tumor dissemination [173,
174]. The tumor-suppressing activity has been observed in
immune-deficient mice and thus does not seem to depend on
the immune system but on other apoptosis stimuli. Another
IAP family member, cIAP2, is affected by the translocation
t(11;18)(q21;q21) that is found in about 50% of marginal cell
lymphomas of the mucosa-associated lymphoid tissue (MALT)
[175]. This suggests a role of cIAP2 in the oncogenesis of
MALT lymphoma.
Tumor cells cannot only resist killing by cytotoxic lympho-
cytes through mechanisms blocking the death receptor path-
way, but also through direct interference with the perforin/
granzyme pathway. The serine protease inhibitor PI-9/SPI-6
that inhibits granzyme B is expressed in a variety of human and
murine tumors. Overexpressions result in resistance of tumor
cells to cytotoxic lymphocytes and to immune escape [176
178].
A different strategy of tumors to escape from the effector
mechanisms of the immune system may be to neutralize or
impair the death-inducing stimuli. Thus, the expression of
soluble receptors that act as decoys for death ligands may
interfere with apoptosis induction via death receptors. To date,
Igney and Krammer Apoptosis and tumor immune escape 911
two distinct soluble receptors, soluble CD95 (sCD95) and
decoy receptor 3 (DcR3), have been shown to inhibit CD95
signaling competitively. sCD95 is expressed in various malig-
nancies, and elevated levels can be found in the sera of cancer
patients [179 182]. High sCD95 serum levels were associated
with poor prognosis in melanoma patients. DcR3 is amplified
genetically in a number of lung and colon carcinomas and is
overepxressed in several adenocarcinomas, glioma cell lines,
and glioblastomas [101, 102, 183, 184]. Ectopic expression of
DcR3 in a rat gliosarcoma model resulted in a decrease of
immune cell infiltration, suggesting an involvement of DcR3 in
immune evasion of malignant glioma [184]. Moreover, a mech-
anism for resistance toward the perforin/granzyme pathway
may be that binding of perforin to the tumor cell membrane is
impaired [185]. Acute myeloid leukemia cells that failed to
bind perforin on their surface were completely resistant toward
NK cell-mediated cytotoxicity.
Tumors can also acquire apoptosis resistance by down-
regulation or inactivation of proapoptotic molecules. In com-
parison to their normal counterparts, some tumor cells show a
decreased expression of the death receptor CD95. This has
been demonstrated for hepatocellular carcinomas, neoplastic
colon epithelium, melanomas, and other tumors [186 190].
Loss of CD95 may contribute to chemoresistance and immune
evasion. Transcriptional mechanisms may account for this.
Oncogenic Ras seems to down-regulate CD95 [191], and in
hepatocellular carcinomas, loss of CD95 expression was ac-
companied by p53 aberrations [190]. Several CD95 gene mu-
tations have been demonstrated in primary samples of myeloma
and T cell leukemia [192194]. The mutations include point
mutations in the cytoplasmic death domain of CD95 and a
deletion leading to a truncated form of the death receptor.
These mutated forms of CD95 may interfere in a dominant-
negative way with apoptosis induction via CD95. In families
with germline CD95 mutations, usually resulting in autoim-
mune lymphoproliferative syndrome, the risk for the develop-
ment of lymphomas is increased [195]. Deletions and muta-
tions of the death receptors TRAIL-R1 and TRAIL-R2 have
also been observed in tumors [196 199]. The frequent deletion
of the chromosomal region 8p21-22 in head and neck cancer
and non-small cell lung cancers affects the TRAIL-R2 gene.
Mutations have been found in the ectodomain or the death
domain of TRAIL-R1 or TRAIL-R2. Further mutations result
in a truncated form or other antiapoptotic forms. Down-regu-
lation or mutation of death receptors may impair tumor immu-
nosurveillance by NK and T cells.
In neuroblastomas with amplification of the oncogene
MYCN, the gene for the initiator caspase-8 is frequently inac-
tivated by DNA methylation and gene deletion [200]. Caspase-
8-deficient neuroblastoma cells are resistant to death receptor-
and doxorubicin-mediated apoptosis.
In certain types of cancer, the proapoptotic Bcl-2 family
member Bax is mutated [201203]. Common mutations com-
prise frameshift mutations leading to loss of expression and
mutations in the BH domains resulting in loss of function.
Tumor cell lines with frameshift mutations are more resistant to
apoptosis. Reduced Bax expression is associated with a poor
response rate to chemotherapy and shorter survival in some
situations [204]. Several mouse studies confirmed the function
912 Journal of Leukocyte Biology Volume 71, June 2002
of Bax as a tumor suppressor [205207]. However, Bax may be
relevant primarily for apoptosis stimuli such as chemotherapy
or p53 and not for apoptosis triggered by the immune system.
Metastatic melanomas often do not express Apaf-1, which
forms an integral part of the apoptosome [208]. A high rate of
allelic loss of the Apaf-1 locus can be observed. The remaining
allele is transcriptionally inactivated by gene methylation.
Apaf-1-negative melanomas fail to respond to chemotherapy, a
situation found commonly in this type of tumor.
Taken together, tumor cells use many mechanisms to ac-
quire apoptosis resistance. Although a direct role of these
mechanisms for immune escape has only been shown in a few
studies, it is likely that apoptosis resistance is not only relevant
for tumorigenesis and resistance to chemotherapy, but also
influences immunosurveillance and immunotherapy.
TUMOR COUNTERATTACK
Tumor cells may not only resist destruction by the immune
system passively. They may also kill tumor-infiltrating lympho-
cytes actively to suppress the anti-tumor immune response, a
phenomenon called tumor counterattack [209 211]. The
weapon tumors may use to delete CD95-sensitive immune
cells is CD95L.
Many publications have supported the idea of tumor coun-
terattack as an immune escape mechanism. CD95/CD95L in-
teractions are discussed as being an important mechanism for
the maintenance of immune privilege. CD95L is expressed in
immune-privileged sites, e.g., the eye and the testis [212214].
Thus, in the eye a high percentage of human corneal trans-
plants are accepted without tissue matching or immunosup-
pressive therapy. In a mouse model, CD95L-positive corneal
allografts were accepted at a rate of about 45%, whereas all
grafts from mice with a mutated, nonfunctional CD95L (gld)
were rejected [214]. Moreover, inflammatory cells entering the
anterior chamber of the eye in response to viral infection
underwent apoptosis dependent on CD95/CD95L interactions
and did not produce any tissue damage [213]. In contrast, viral
infection in gld mice, which lack functional CD95L, resulted in
inflammation and invasion of ocular tissue by cells without
signs of apoptosis. CD95-mediated apoptosis of lymphoid cells
was necessary for tolerance induction following antigen injec-
tion into the anterior chamber of the eye [215].
Data along similar lines were also found with murine testis
grafts. Grafts expressing wild-type CD95L survived indefinitely
when transplanted under the kidney capsule of allogeneic
animals, whereas testis grafts from gld mice were rejected
[216]. However, attempts to reproduce these observations by
another group were not successful [217]. Because neurons and
astrocytes also express CD95L [212, 218, 219], immune priv-
ilege in the central nervous system may also involve CD95L.
These findings suggest that CD95L may be used to render a
transplanted tissue an immune privileged site. Indeed, it was
demonstrated that cotransplantation of syngeneic CD95L-
transfected myoblasts protected allogeneic islets of Langerhans
grafts from immune rejection [220]. However, other authors
have found exactly opposite results using a similar technology
[221]. Numerous studies have shown that tumor counterattack
http://www.jleukbio.org
may be a relevant immune escape mechanism of tumors. Many
CD95-resistant tumors express CD95L constitutively or after
induction by chemotherapy [211, 218, 222227]. Such tumor
cells killed CD95-positive, apoptosis-sensitive cells in vitro.
Moreover, apoptosis of tumor-infiltrating lymphocytes has been
found in situ within CD95L-expressing human tumors [186,
224]. In esophageal cancer, the number of infiltrated lympho-
cytes was reduced concomitantly with increased lymphocyte
apoptosis within CD95L-expressing areas of the tumors [223].
Various animal models have been used to demonstrate the
ability of CD95L expressed on tumors to down-regulate anti-
tumor immune responses. Tumor growth of a subcutaneously
injected CD95L-positive murine melanoma cell line was
slightly faster in wild-type or gld mice than in mice with a
mutated or down-regulated CD95 receptor (lpr mice) [224].
Another study in syngeneic mice showed that growth of tumors
of murine CD95L-transfected cells was significantly better than
that of control cells when implanted under the kidney capsule
[228]. Immunosuppression in vivo was directly demonstrated
in allogeneic mice injected with CD95L-transfected colon car-
cinoma cells. Alloantibodies were virtually completely abol-
ished and allospecific cytotoxic T lymphocytes and helper T
cells were reduced [229].
Despite the wealth of data accumulated in support of the
tumor counterattack hypothesis, many contradictory studies
have also been published [230]. In contrast to the above
findings, it has been shown that CD95L expression by grafts or
by tumor cells targeted the cells for rapid destruction by
neutrophils. CD95L expression on pancreatic islets trans-
planted into allogeneic hosts resulted in a massive infiltration
of neutrophils and in islet destruction. Similarly, transgenic
mice expressing CD95L in pancreatic cells developed a
massive infiltration of neutrophils and diabetes [231]. CD95L-
expressing islet cells transplanted under the kidney capsule
of syngeneic or allogeneic animals were not protected from
rejection [232, 233]. In addition, CD95L-expressing hearts
from transgenic mice transplanted into sygneneic and alloge-
neic recipients were more rapidly rejected than control grafts
and showed massive neutrophil infiltration as early as 1 day
after transplantation [234].
Various studies have shown that the overexpression of
CD95L in murine tumor cells resistant to CD95-mediated
apoptosis did not affect growth in vitro, but caused rejection by
neutrophils in vivo [235239]. CD8 T cell-mediated protec-
tive immunity against subsequent challenge with the parental
tumor cells was elicited. Rejection of the CD95L-expressing
tumor has even been observed in the study mentioned above,
demonstrating immunosuppression by the tumor in an alloge-
neic mouse model [229]. In addition, when control tumor cells
were co-implanted with the CD95L-expressing cells at the
same site, a bystander rejection of the CD95L-negative cells
was found [236,237]. Infection of a subcutaneously growing
CD95-negative tumor by an adenoviral vector encoding CD95L
resulted in rapid elimination of the tumor [240]. These studies
indicate that CD95L has a proinflammatory function and that
gene transfer of CD95L may be used in tumor eradication.
Several mechanisms for the recruitment of the graft- or
tumor-rejecting neutrophils have been proposed. Two studies
suggested that soluble CD95L is directly chemotactic for neu-
trophils implicating that soluble CD95L promotes rejection of
CD95L-expressing grafts [241, 242]. However, others have not
found a chemotactic activity of soluble CD95L in vitro or in
vivo [235, 243]. Moreover, tumor cells expressing only soluble
CD95L did not elicit a neutrophilic response [243, 244]. Con-
versely, tumor cells expressing a noncleavable membrane-
bound form of CD95L were rapidly rejected [243245]. The
extent of inflammation induced by the various transfectants
seemed to correlate with the cytotoxic activity of CD95L.
Alternatively, it has been suggested that CD95L acts on sur-
rounding cells to induce the production of granulocyte che-
moattractants. Thus, it has been demonstrated that CD95L
induces the processing and release of IL-1, which may be
responsible for the infiltration by neutrophils [246]. CD95L
may act on resident macrophages, leading to increased pro-
duction of IL-1 and macrophage inflammatory proteins [247].
Moreover, engagement of CD95 on dendritic cells may induce
the secretion of proinflammatory cytokines [248]. These data
support an indirect mechanism for the inflammatory effect of
CD95L. Nevertheless, the exact mechanism of how neutrophils
are recruited to CD95L-expressing tumors and how CD95L-
expressing tumors are rejected is still unclear.
Many studies of tumor counterattack have been published,
but the results are contradictory, and therefore do not clarify
whether tumor counterattack is a relevant immune escape
mechanism in vivo. No study has demonstrated conclusively
that a tumor (or graft), by CD95L expression, deleted anti-
tumor-specific lymphocytes, escaped the immune response,
and thus had a growth advantage in vivo.
Additional factors may influence the outcome of CD95L
expression on tumors in vivo and thus may account for the
controversial situation. Level and time-point of CD95L expres-
sion may be particularly relevant. In most animal experiments,
CD95L-transfected tumor cell lines have been used. These
cells may express different, probably higher levels of CD95L
than naturally occurring tumors. Overexpression of CD95L
may lead to rejection by neutrophils, whereas physiological
levels may not induce neutrophilic infiltration but may still
suffice to delete anti-tumor lymphocytes. Moreover, tumors
growing in situ may express CD95L at late stages of tumori-
genesis, e.g., after induction by chemotherapy [249, 250].
Therefore, the time-point of CD95L expression may directly
influence tumor counterattack. Furthermore, the sensitivity of
T cells to CD95-mediated apoptosis varies considerably with
respect to the activation status of the T cells [84, 89, 251253].
Therefore, it may be crucial when T cells encounter CD95L
and how the T cells have been activated and costimulated. The
consequences of CD95L expression may also depend on the
microenvironment. It has been suggested that TGF- is nec-
essary as a cofactor for promoting immunologic tolerance [254].
CD95L-expressing tumor cells were rejected by neutrophils
when injected subcutaneously. When TGF- was simulta-
neously provided to the subcutaneous sites, protection against
tumor rejection was observed. However, it is not clear whether
in vivo TGF- and CD95L together are necessary for immune
escape and whether CD95L has an immunosuppressive effect
on T cells in the presence of TGF-.
Despite the controversial situation concerning CD95L and
tumor counterattack, other molecules have also been impli-
Igney and Krammer Apoptosis and tumor immune escape 913
cated in T cell deletion by tumors. Thus, the death ligand
TRAIL has been suggested to suppress tumor-specific T cell
responses in a similar manner as CD95L [255]. Moreover, the
membrane protein RCAS1 [67] not only inhibits proliferation,
but also induces apoptosis in T cells in vitro. Chemokine
production by tumor cells may also sensitize T cells to apo-
ptosis [256].
CONCLUSIONS
Research in tumor immunology has provided a wealth of in-
formation about the interactions between tumors and the im-
mune system. Many of these interactions are now not only
known on a cellular, but also on a molecular level. Despite this
knowledge, cancer immunotherapy still is not an established
treatment in the clinic. Many approaches may fail because
tumors use multiple mechanisms to become resistant to apo-
ptosis or to counterattack the immune system. These mecha-
nisms render tumor cells insensitive to the effector mecha-
nisms of the immune system, yet the significance for immune
escape has only been shown in few studies.
It is the present aim of research in this area to understand
these events further and to use this insight to resensitize tumor
cells to apoptosis. Recently, it has been shown that low doses
of chemotherapy or irradiation sensitized resistant cells to
TRAIL-induced apoptosis in vitro and in vivo [257, 258].
Therefore, these treatments might enhance the susceptibility of
a tumor to immune attack. A future therapeutic strategy may
also involve down-regulation of antiapoptotic molecules such
as FLIP or Bcl-2 by antisense oligonucleotides or dsRNA
interference [259, 260]. However, a macroscopic tumor is
heterogeneous, and different cells within the tumor may also
use different immune escape mechanisms including apoptosis
resistance, impaired antigen presentation, secretion of immu-
nosuppressive factors, and other strategies (Table 1). Moreover,
multiple mechanisms may develop in a single tumor cell.
Therefore, it is questionable whether a single, predominant
immune-escape mechanism can be identified in a tumor and
whether therapeutic targeting of one mechanism alone is prom-
ising. Deeper insight into the molecular mechanisms underly-
ing tumor immune escape may finally lead to novel therapeutic
approaches that will be used for the benefit of cancer patients.
ACKNOWLEDGMENTS
This work was supported by grants from the Deutsche For-
schungsgemeinschaft, the European Community, the Deutsche
Krebshilfe, the Sander Stiftung, the Tumor Center Heidelberg/
Mannheim, the BMBF, and the German-Israeli Cooperation in
Cancer Research.
REFERENCES
1. Smyth, M. J., Godfrey, D. I., Trapani, J. A. (2001) A fresh look at tumor
immunosurveillance and immunotherapy. Nat. Immunol. 2, 293299.
914 Journal of Leukocyte Biology Volume 71, June 2002
2. Sogn, J. A. (1998) Tumor immunology: the glass is half full. Immunity 9,
757763.
3. Rosenberg, S. A. (2001) Progress in human tumour immunology and
immunotherapy. Nature 411, 380 384.
4. Houghton, A. N., Gold, J. S., Blachere, N. E. (2001) Immunity against
cancer: lessons learned from melanoma. Curr. Opin. Immunol. 13, 134
140.
5. Boon, T., van der Bruggen, P. (1996) Human tumor antigens recognized
by T lymphocytes. J. Exp. Med. 183, 725729.
6. Lee, P. P., Yee, C., Savage, P. A., Fong, L., Brockstedt, D., Weber, J. S.,
Johnson, D., Swetter, S., Thompson, J., Greenberg, P. D., Roederer, M.,
Davis, M. M. (1999) Characterization of circulating T cells specific for
tumor-associated antigens in melanoma patients. Nat. Med. 5, 677 685.
7. Wortzel, R. D., Philipps, C., Schreiber, H. (1983) Multiple tumour-
specific antigens expressed on a single tumour cell. Nature 304, 165
167.
8. Burnet, F. M. (1970) The concept of immunological surveillance. Prog.
Exp. Tumor Res. 13, 127.
9. Fuchs, E. J., Matzinger, P. (1996) Is cancer dangerous to the immune
system? Semin. Immunol. 8, 271280.
10. Watzl, C., Long, E. O. (2000) Exposing tumor cells to killer cell attack.
Nat. Med. 6, 867 868.
11. Pardoll, D. M. (2001) Immunology. Stress, NK receptors, and immune
surveillance. Science 294, 534 536.
12. Moretta, A., Biassoni, R., Bottino, C., Mingari, M. C., Moretta, L. (2000)
Natural cytotoxicity receptors that trigger human NK-cell-mediated cy-
tolysis. Immunol. Today 21, 228 234.
13. Cerwenka, A., Bakker, A. B., McClanahan, T., Wagner, J., Wu, J.,
Phillips, J. H., Lanier, L. L. (2000) Retinoic acid early inducible genes
define a ligand family for the activating NKG2D receptor in mice.
Immunity 12, 721727.
14. Bauer, S., Groh, V., Wu, J., Steinle, A., Phillips, J. H., Lanier, L. L.,
Spies, T. (1999) Activation of NK cells and T cells by NKG2D, a receptor
for stress-inducible MICA. Science 285, 727729.
15. Wu, J., Song, Y., Bakker, A. B., Bauer, S., Spies, T., Lanier, L. L.,
Phillips, J. H. (1999) An activating immunoreceptor complex formed by
NKG2D and DAP10. Science 285, 730 732.
16. Lollini, P. L., Forni, G. (1999) Specific and nonspecific immunity in the
prevention of spontaneous tumours. Immunol Today 20, 347350.
17. Di Carlo, E., Forni, G., Lollini, P., Colombo, M. P., Modesti, A., Musiani,
P. (2001) The intriguing role of polymorphonuclear neutrophils in anti-
tumor reactions. Blood 97, 339 345.
18. Elgert, K. D., Alleva, D. G., Mullins, D. W. (1998) Tumor-induced
immune dysfunction: the macrophage connection. J. Leukoc. Biol. 64,
275290.
19. Bonnotte, B., Larmonier, N., Favre, N., Fromentin, A., Moutet, M.,
Martin, M., Gurbuxani, S., Solary, E., Chauffert, B., Martin, F. (2001)
Identification of tumor-infiltrating macrophages as the killers of tumor
cells after immunization in a rat model system. J. Immunol. 167, 5077
5083.
20. Lin, E. Y., Nguyen, A. V., Russell, R. G., Pollard, J. W. (2001) Colony-
stimulating factor 1 promotes progression of mammary tumors to malig-
nancy. J. Exp. Med. 193, 727740.
21. Chen, L. (1998) Immunological ignorance of silent antigens as an expla-
nation of tumor evasion. Immunol. Today 19, 2730.
22. Ochsenbein, A. F., Klenerman, P., Karrer, U., Ludewig, B., Pericin, M.,
Hengartner, H., Zinkernagel, R. M. (1999) Immune surveillance against
a solid tumor fails because of immunological ignorance. Proc. Natl. Acad.
Sci. USA 96, 22332238.
23. Ochsenbein, A. F., Sierro, S., Odermatt, B., Pericin, M., Karrer, U.,
Hermans, J., Hemmi, S., Hengartner, H., Zinkernagel, R. M. (2001)
Roles of tumour localization, second signals and cross priming in cyto-
toxic T-cell induction. Nature 411, 1058 1064.
24. Rocha, G., Deschenes, J., Rowsey, J. J. (1998) The immunology of
corneal graft rejection. Crit. Rev. Immunol. 18, 305325.
25. Onrust, S. V., Hartl, P. M., Rosen, S. D., Hanahan, D. (1996) Modulation
of L-selectin ligand expression during an immune response accompany-
ing tumorigenesis in transgenic mice. J. Clin. Investig. 97, 54 64.
26. Piali, L., Fichtel, A., Terpe, H. J., Imhof, B. A., Gisler, R. H. (1995)
Endothelial vascular cell adhesion molecule 1 expression is suppressed
by melanoma and carcinoma. J. Exp. Med. 181, 811 816.
27. Ishido, S., Choi, J. K., Lee, B. S., Wang, C., DeMaria, M., Johnson, R. P.,
Cohen, G. B., Jung, J. U. (2000) Inhibition of natural killer cell-mediated
cytotoxicity by Kaposis sarcoma-associated herpesvirus K5 protein.
Immunity 13, 365374.
28. Singh, S., Ross, S. R., Acena, M., Rowley, D. A., Schreiber, H. (1992)
Stroma is critical for preventing or permitting immunological destruction
of antigenic cancer cells. J. Exp. Med. 175, 139 146.
http://www.jleukbio.org
29. Wick, M., Dubey, P., Koeppen, H., Siegel, C. T., Fields, P. E., Chen, L.,
Bluestone, J. A., Schreiber, H. (1997) Antigenic cancer cells grow
progressively in immune hosts without evidence for T cell exhaustion or
systemic anergy. J. Exp. Med. 186, 229 238.
30. Antonia, S. J., Extermann, M., Flavell, R. A. (1998) Immunologic non-
responsiveness to tumors. Crit. Rev. Oncog. 9, 35 41.
31. Villunger, A., Strasser, A. (1999) The great escape: is immune evasion
required for tumor progression? Nat. Med. 5, 874 875.
32. Gilboa, E. (1999) How tumors escape immune destruction and what we
can do about it. Cancer Immunol. Immunother. 48, 382385.
33. Maeurer, M. J., Gollin, S. M., Martin, D., Swaney, W., Bryant, J., Castelli,
C., Robbins, P., Parmiani, G., Storkus, W. J., Lotze, M. T. (1996) Tumor
escape from immune recognition: lethal recurrent melanoma in a patient
associated with downregulation of the peptide transporter protein TAP-1
and loss of expression of the immunodominant MART-1/Melan-A anti-
gen. J. Clin. Investig. 98, 16331641.
34. Kim, U., Baumler, A., Carruthers, C., Bielat, K. (1975) Immunological
escape mechanism in spontaneously metastasizing mammary tumors.
Proc. Natl. Acad. Sci. USA 72, 10121016.
35. Vasmel, W. L., Sijts, E. J., Leupers, C. J., Matthews, E. A., Melief, C. J.
(1989) Primary virus-induced lymphomas evade T cell immunity by
failure to express viral antigens. J. Exp. Med. 169, 12331254.
36. Stackpole, C. W., Cremona, P., Leonard, C., Stremmel, P. (1980) Anti-
genic modulation as a mechanism for tumor escape from immune de-
struction: identification of modulation-positive and modulation-negative
mouse lymphomas with xenoantisera to murine leukemia virus gp70.
J. Immunol. 125, 17151723.
37. Hicklin, D. J., Marincola, F. M., Ferrone, S. (1999) HLA class I antigen
downregulation in human cancers: T-cell immunotherapy revives an old
story. Mol. Med. Today 5, 178 186.
38. Garrido, F., Ruiz-Cabello, F., Cabrera, T., Perez-Villar, J. J., Lopez-
Botet, M., Duggan-Keen, M., Stern, P. L. (1997) Implications for immu-
nosurveillance of altered HLA class I phenotypes in human tumours.
Immunol. Today 18, 89 95.
39. Bicknell, D. C., Rowan, A., Bodmer, W. F. (1994) Beta 2-microglobulin
gene mutations: a study of established colorectal cell lines and fresh
tumors. Proc. Natl. Acad. Sci. USA 91, 4751 4756.
40. Seung, S., Urban, J. L., Schreiber, H. (1993) A tumor escape variant that
has lost one major histocompatibility complex class I restriction element
induces specific CD8 T cells to an antigen that no longer serves as a
target. J. Exp. Med. 178, 933940.
41. Natali, P. G., Nicotra, M. R., Bigotti, A., Venturo, I., Marcenaro, L.,
Giacomini, P., Russo, C. (1989) Selective changes in expression of HLA
class I polymorphic determinants in human solid tumors. Proc. Natl.
Acad. Sci. USA 86, 6719 6723.
42. Blanchet, O., Bourge, J. F., Zinszner, H., Israel, A., Kourilsky, P.,
Dausset, J., Degos, L., Paul, P. (1992) Altered binding of regulatory
factors to HLA class I enhancer sequence in human tumor cell lines
lacking class I antigen expression. Proc. Natl. Acad. Sci. USA 89,
3488 3492.
43. Bosshart, H., Jarrett, R. F. (1998) Deficient major histocompatibility
complex class II antigen presentation in a subset of Hodgkins disease
tumor cells. Blood 92, 22522259.
44. Farrell, H. E., Vally, H., Lynch, D. M., Fleming, P., Shellam, G. R.,
Scalzo, A. A., Davis-Poynter, N. J. (1997) Inhibition of natural killer cells
by a cytomegalovirus MHC class I homologue in vivo. Nature 386,
510 514.
45. Cretney, E., Degli-Esposti, M. A., Densley, E. H., Farrell, H. E., Davis-
Poynter, N. J., Smyth, M. J. (1999) m144, a murine cytomegalovirus
(MCMV)-encoded major histocompatibility complex class I homologue,
confers tumor resistance to natural killer cell-mediated rejection. J. Exp.
Med. 190, 435 444.
46. Seliger, B., Maeurer, M. J., Ferrone, S. (1997) TAP offtumors on.
Immunol. Today 18, 292299.
47. Restifo, N. P., Esquivel, F., Kawakami, Y., Yewdell, J. W., Mule, J. J.,
Rosenberg, S. A., Bennink, J. R. (1993) Identification of human cancers
deficient in antigen processing. J. Exp. Med. 177, 265272.
48. Rotem-Yehudar, R., Groettrup, M., Soza, A., Kloetzel, P. M., Ehrlich, R.
(1996) LMP-associated proteolytic activities and TAP-dependent peptide
transport for class 1 MHC molecules are suppressed in cell lines trans-
formed by the highly oncogenic adenovirus 12. J. Exp. Med. 183,
499 514.
49. Cromme, F. V., Airey, J., Heemels, M. T., Ploegh, H. L., Keating, P. J.,
Stern, P. L., Meijer, C. J., Walboomers, J. M. (1994) Loss of transporter
protein, encoded by the TAP-1 gene, is highly correlated with loss of
HLA expression in cervical carcinomas. J. Exp. Med. 179, 335340.
50. Johnsen, A. K., Templeton, D. J., Sy, M., Harding, C. V. (1999) Defi-
ciency of transporter for antigen presentation (TAP) in tumor cells allows
Igney and Krammer Apoptosis and tumor immune escape
evasion of immune surveillance and increases tumorigenesis. J. Immu-
nol. 163, 4224 4231.
51. Ossendorp, F., Eggers, M., Neisig, A., Ruppert, T., Groettrup, M., Sijts,
A., Mengede, E., Kloetzel, P. M., Neefjes, J., Koszinowski, U., Melief, C.
(1996) A single residue exchange within a viral CTL epitope alters
proteasome-mediated degradation resulting in lack of antigen presenta-
tion. Immunity 5, 115124.
52. Chouaib, S., Asselin-Paturel, C., Mami-Chouaib, F., Caignard, A., Blay,
J. Y. (1997) The host-tumor immune conflict: from immunosuppression to
resistance and destruction. Immunol. Today 18, 493 497.
53. Loeffler, C. M., Smyth, M. J., Longo, D. L., Kopp, W. C., Harvey, L. K.,
Tribble, H. R., Tase, J. E., Urba, W. J., Leonard, A. S., Young, H. A., et.
al. (1992) Immunoregulation in cancer-bearing hosts. Down-regulation of
gene expression and cytotoxic function in CD8 T cells. J. Immunol.
149, 949 956.
54. Pasche, B. (2001) Role of transforming growth factor beta in cancer.
J. Cell. Physiol. 186, 153168.
55. Gorelik, L., Flavell, R. A. (2001) Immune-mediated eradication of tu-
mors through the blockade of transforming growth factor-beta signaling in
T cells. Nat. Med. 7, 1118 1122.
56. Tada, T., Ohzeki, S., Utsumi, K., Takiuchi, H., Muramatsu, M., Li, X. F.,
Shimizu, J., Fujiwara, H., Hamaoka, T. (1991) Transforming growth
factor-beta-induced inhibition of T cell function. Susceptibility differ-
ence in T cells of various phenotypes and functions and its relevance to
immunosuppression in the tumor-bearing state. J. Immunol. 146, 1077
1082.
57. Ranges, G. E., Figari, I. S., Espevik, T., Palladino Jr., M. A. (1987)
Inhibition of cytotoxic T cell development by transforming growth factor
beta and reversal by recombinant tumor necrosis factor alpha. J. Exp.
Med. 166, 991998.
58. Torre-Amione, G., Beauchamp, R. D., Koeppen, H., Park, B. H., Schre-
iber, H., Moses, H. L., Rowley, D. A. (1990) A highly immunogenic
tumor transfected with a murine transforming growth factor type beta 1
cDNA escapes immune surveillance. Proc. Natl. Acad. Sci. USA 87,
1486 1490.
59. Gabrilovich, D. I., Chen, H. L., Girgis, K. R., Cunningham, H. T., Meny,
G. M., Nadaf, S., Kavanaugh, D., Carbone, D. P. (1996) Production of
vascular endothelial growth factor by human tumors inhibits the func-
tional maturation of dendritic cells. Nat. Med. 2, 1096 1103.
60. Plescia, O. J., Smith, A. H., Grinwich, K. (1975) Subversion of immune
system by tumor cells and role of prostaglandins. Proc. Natl. Acad. Sci.
USA 72, 1848 1851.
61. Young, M. R., Knies, S. (1984) Prostaglandin E production by Lewis lung
carcinoma: mechanism for tumor establishment in vivo. J. Natl. Cancer
Inst. 72, 919 922.
62. McLemore, T. L., Hubbard, W. C., Litterst, C. L., Liu, M. C., Miller, S.,
McMahon, N. A., Eggleston, J. C.,Boyd, M. R. (1988) Profiles of prosta-
glandin biosynthesis in normal lung and tumor tissue from lung cancer
patients. Cancer Res. 48, 3140 3147.
63. Matsuda, M., Salazar, F., Petersson, M., Masucci, G., Hansson, J., Pisa,
P., Zhang, Q. J., Masucci, M. G., Kiessling, R. (1994) Interleukin 10
pretreatment protects target cells from tumor- and allo-specific cytotoxic
T cells and downregulates HLA class I expression. J. Exp. Med. 180,
23712376.
64. Oghiso, Y., Yamada, Y., Ando, K., Ishihara, H., Shibata, Y. (1993)
Differential induction of prostaglandin E2-dependent and -independent
immune suppressor cells by tumor-derived GM-CSF and M-CSF. J.
Leukoc. Biol. 53, 86 92.
65. Sotomayor, E. M., Fu, Y. X., Lopez-Cepero, M., Herbert, L., Jimenez,
J. J., Albarracin, C., Lopez, D. M. (1991) Role of tumor-derived cytokines
on the immune system of mice bearing a mammary adenocarcinoma. II.
Down-regulation of macrophage-mediated cytotoxicity by tumor-derived
granulocyte-macrophage colony-stimulating factor. J. Immunol. 147,
2816 2823.
66. McKallip, R., Li, R., Ladisch, S. (1999) Tumor gangliosides inhibit the
tumor-specific immune response. J. Immunol. 163, 3718 3726.
67. Nakashima, M., Sonoda, K., Watanabe, T. (1999) Inhibition of cell
growth and induction of apoptotic cell death by the human tumor-
associated antigen RCAS1. Nat. Med. 5, 938 942.
68. Kiessling, R., Wasserman, K., Horiguchi, S., Kono, K., Sjoberg, J., Pisa,
P., Petersson, M. (1999) Tumor-induced immune dysfunction. Cancer
Immunol. Immunother. 48, 353362.
69. Arnold, B., Schonrich, G., Hammerling, G. J. (1993) Multiple levels of
peripheral tolerance. Immunol. Today 14, 1214.
70. Ganss, R., Limmer, A., Sacher, T., Arnold, B., Hammerling, G. J. (1999)
Autoaggression and tumor rejection: it takes more than self-specific
T-cell activation. Immunol. Rev. 169, 263272.
915
71. Chen, L., Ashe, S., Brady, W. A., Hellstrom, I., Hellstrom, K. E.,
Ledbetter, J. A., McGowan, P., Linsley, P. S. (1992) Costimulation of
antitumor immunity by the B7 counterreceptor for the T lymphocyte
molecules CD28 and CTLA-4. Cell 71, 10931102.
72. Guinan, E. C., Gribben, J. G., Boussiotis, V. A., Freeman, G. J., Nadler,
L. M. (1994) Pivotal role of the B7:CD28 pathway in transplantation
tolerance and tumor immunity. Blood 84, 32613282.
73. Staveley-OCarroll, K., Sotomayor, E., Montgomery, J., Borrello, I.,
Hwang, L., Fein, S., Pardoll, D., Levitsky, H. (1998) Induction of
antigen-specific T cell anergy: an early event in the course of tumor
progression. Proc. Natl. Acad. Sci. USA 95, 1178 1183.
74. Becker, J. C., Brabletz, T., Czerny, C., Termeer, C., Brocker, E. B. (1993)
Tumor escape mechanisms from immunosurveillance: induction of un-
responsiveness in a specific MHC-restricted CD4 human T cell clone
by the autologous MHC class II melanoma. Int. Immunol. 5, 1501
1508.
75. Shrikant, P., Khoruts, A., Mescher, M. F. (1999) CTLA-4 blockade
reverses CD8 T cell tolerance to tumor by a CD4 T cell- and
IL-2-dependent mechanism. Immunity 11, 483 493.
76. Maeda, H., Shiraishi, A. (1996) TGF-beta contributes to the shift toward
Th2-type responses through direct and IL-10-mediated pathways in
tumor-bearing mice. J. Immunol. 156, 7378.
77. Qin, Z., Richter, G., Schuler, T., Ibe, S., Cao, X., Blankenstein, T. (1998)
B cells inhibit induction of T cell-dependent tumor immunity. Nat. Med.
4, 627 630.
78. Sakaguchi, S. (2000) Regulatory T cells: key controllers of immunologic
self-tolerance. Cell 101, 455 458.
79. Matsui, S., Ahlers, J. D., Vortmeyer, A. O., Terabe, M., Tsukui, T.,
Carbone, D. P., Liotta, L. A., Berzofsky, J. A. (1999) A model for CD8
CTL tumor immunosurveillance and regulation of tumor escape by CD4
T cells through an effect on quality of CTL. J. Immunol. 163, 184 193.
80. Takahashi, K., Ono, K., Hirabayashi, Y., Taniguchi, M. (1988) Escape
mechanisms of melanoma from immune system by soluble melanoma
antigen. J. Immunol. 140, 3244 3248.
81. Russell, J. H., White, C. L., Loh, D. Y., Meleedy-Rey, P. (1991) Recep-
tor-stimulated death pathway is opened by antigen in mature T cells.
Proc. Natl. Acad. Sci. USA 88, 21512155.
82. Radvanyi, L. G., Mills, G. B., Miller, R. G. (1993) Religation of the T cell
receptor after primary activation of mature T cells inhibits proliferation
and induces apoptotic cell death. J. Immunol. 150, 5704 5715.
83. Martin, D. R., Miller, R. G. (1989) In vivo administration of histoincom-
patible lymphocytes leads to rapid functional deletion of cytotoxic T
lymphocyte precursors. J. Exp. Med. 170, 679 690.
84. Kirchhoff, S., Muller, W. W., Krueger, A., Schmitz, I., Krammer, P. H.
(2000) TCR-mediated up-regulation of c-FLIPshort correlates with re-
sistance toward CD95-mediated apoptosis by blocking death-inducing
signaling complex activity. J. Immunol. 165, 6293 6300.
85. Singer, G. G., Abbas, A. K. (1994) The fas antigen is involved in
peripheral but not thymic deletion of T lymphocytes in T cell receptor
transgenic mice. Immunity 1, 365371.
86. Dhein, J., Walczak, H., Baumler, C., Debatin, K. M., Krammer, P. H.
(1995) Autocrine T-cell suicide mediated by APO-1/(Fas/CD95). Nature
373, 438 441.
87. Brunner, T., Mogil, R. J., LaFace, D., Yoo, N. J., Mahboubi, A., Echev-
erri, F., Martin, S. J., Force, W. R., Lynch, D. H., Ware, C. F., et al.
(1995) Cell-autonomous Fas (CD95)/Fas-ligand interaction mediates ac-
tivation-induced apoptosis in T-cell hybridomas. Nature 373, 441 444.
88. Li-Weber, M., Laur, O., Hekele, A., Coy, J., Walczak, H., Krammer,
P. H. (1998) A regulatory element in the CD95 (APO-1/Fas) ligand
promoter is essential for responsiveness to TCR-mediated activation.
Eur. J. Immunol. 28, 23732383.
89. Kirchhoff, S., Muller, W. W., Li-Weber, M., Krammer, P. H. (2000)
Up-regulation of c-FLIPshort and reduction of activation-induced cell
death in CD28-costimulated human T cells. Eur. J. Immunol. 30, 2765
2774.
90. Bogen, B. (1996) Peripheral T cell tolerance as a tumor escape mecha-
nism: deletion of CD4 T cells specific for a monoclonal immunoglob-
ulin idiotype secreted by a plasmacytoma. Eur. J. Immunol. 26, 2671
2679.
91. Lauritzsen, G. F., Hofgaard, P. O., Schenck, K., Bogen, B. (1998) Clonal
deletion of thymocytes as a tumor escape mechanism. Int. J. Cancer 78,
216 222.
92. Shresta, S., Pham, C. T., Thomas, D. A., Graubert, T. A., Ley, T. J. (1998)
How do cytotoxic lymphocytes kill their targets? Curr. Opin. Immunol.
10, 581587.
93. Rouvier, E., Luciani, M. F., Golstein, P. (1993) Fas involvement in
Ca(2)-independent T cell-mediated cytotoxicity. J. Exp. Med. 177,
195200.
916 Journal of Leukocyte Biology Volume 71, June 2002
94. Walsh, C. M., Glass, A. A., Chiu, V., Clark, W. R. (1994) The role of the
Fas lytic pathway in a perforin-less CTL hybridoma. J. Immunol. 153,
2506 2514.
95. Li, J. H., Rosen, D., Ronen, D., Behrens, C. K., Krammer, P. H., Clark,
W. R., Berke, G. (1998) The regulation of CD95 ligand expression and
function in CTL. J. Immunol. 161, 39433949.
96. Smyth, M. J., Cretney, E., Takeda, K., Wiltrout, R. H., Sedger, L. M.,
Kayagaki, N., Yagita, H., Okumura, K. (2001) Tumor necrosis factor-
related apoptosis-inducing ligand (TRAIL) contributes to interferon gamma-
dependent natural killer cell protection from tumor metastasis. J. Exp.
Med. 193, 661 670.
97. Takeda, K., Hayakawa, Y., Smyth, M. J., Kayagaki, N., Yamaguchi, N.,
Kakuta, S., Iwakura, Y., Yagita, H., Okumura, K. (2001) Involvement of
tumor necrosis factor-related apoptosis-inducing ligand in surveillance of
tumor metastasis by liver natural killer cells. Nat. Med. 7, 94 100.
98. Takeda, K., Smyth, M. J., Cretney, E., Hayakawa, Y., Kayagaki, N.,
Yagita, H., Okumura, K. (2002) Critical role for tumor necrosis factor-
related apoptosis-inducing ligand in immune surveillance against tumor
development. J. Exp. Med. 195, 161169.
99. Krammer, P. H. (1999) CD95(APO-1/Fas)-mediated apoptosis: live and
let die. Adv. Immunol. 71, 163210.
100. Schmitz, I., Kirchhoff, S., Krammer, P. H. (2000) Regulation of death
receptor-mediated apoptosis pathways. Int. J. Biochem. Cell Biol. 32,
11231136.
101. Pitti, R. M., Marsters, S. A., Lawrence, D. A., Roy, M., Kischkel, F. C.,
Dowd, P., Huang, A., Donahue, C. J., Sherwood, S. W., Baldwin, D. T.,
Godowski, P. J., Wood, W. I., Gurney, A. L., Hillan, K. J., Cohen, R. L.,
Goddard, A. D., Botstein, D., Ashkenazi, A. (1998) Genomic amplifica-
tion of a decoy receptor for Fas ligand in lung and colon cancer. Nature
396, 699 703.
102. Yu, K. Y., Kwon, B., Ni, J., Zhai, Y., Ebner, R., Kwon, B. S. (1999) A
newly identified member of tumor necrosis factor receptor superfamily
(TR6) suppresses LIGHT-mediated apoptosis. J. Biol. Chem. 274,
1373313736.
103. Kischkel, F. C., Hellbardt, S., Behrmann, I., Germer, M., Pawlita, M.,
Krammer, P. H., Peter, M. E. (1995) Cytotoxicity-dependent APO-1
(Fas/CD95)-associated proteins form a death-inducing signaling complex
(DISC) with the receptor. EMBO J. 14, 5579 5588.
104. Muzio, M., Chinnaiyan, A. M., Kischkel, F. C., ORourke, K.,
Shevchenko, A., Ni, J., Scaffidi, C., Bretz, J. D., Zhang, M., Gentz, R.,
Mann, M., Krammer, P. H., Peter, M. E., Dixit, V. M. (1996) FLICE, a
novel FADD-homologous ICE/CED-3-like protease, is recruited to the
CD95 (Fas/APO-1) deathinducing signaling complex. Cell 85, 817
827.
105. Sprick, M. R., Weigand, M. A., Rieser, E., Rauch, C. T., Juo, P., Blenis,
J., Krammer, P. H., Walczak, H. (2000) FADD/MORT1 and caspase-8
are recruited to TRAIL receptors 1 and 2 and are essential for apoptosis
mediated by TRAIL receptor 2. Immunity 12, 599 609.
106. Kischkel, F. C., Lawrence, D. A., Chuntharapai, A., Schow, P., Kim,
K. J., Ashkenazi, A. (2000) Apo2L/TRAIL-dependent recruitment of
endogenous FADD and caspase-8 to death receptors 4 and 5. Immunity
12, 611 620.
107. Kischkel, F. C., Lawrence, D. A., Tinel, A., LeBlanc, H., Virmani, A.,
Schow, P., Gazdar, A., Blenis, J., Arnott, D., Ashkenazi, A. (2001) Death
receptor recruitment of endogenous caspase-10 and apoptosis initiation
in the absence of caspase-8. J. Biol. Chem. 276, 46639 46646.
108. Scaffidi, C., Fulda, S., Srinivasan, A., Friesen, C., Li, F., Tomaselli, K. J.,
Debatin, K. M., Krammer, P. H., Peter, M. E. (1998) Two CD95 (APO-
1/Fas) signaling pathways. EMBO J. 17, 16751687.
109. Zamzami, N., Kroemer, G. (2001) The mitochondrion in apoptosis: how
Pandoras box opens. Nat. Rev. Mol. Cell. Biol. 2, 6771.
110. Martinou, J. C., Green, D. R. (2001) Breaking the mitochondrial barrier.
Nat. Rev. Mol. Cell. Biol. 2, 63 67.
111. Rathmell, J. C., Thompson, C. B. (1999) The central effectors of cell
death in the immune system. Annu. Rev. Immunol. 17, 781 828.
112. Krueger, A., Baumann, S., Krammer, P. H., Kirchhoff, S. (2001) FLICE-
inhibitory proteins: regulators of death receptor-mediated apoptosis. Mol.
Cell. Biol. 21, 8247 8254.
113. Deveraux, Q. L., Reed, J. C. (1999) IAP family proteinssuppressors of
apoptosis. Genes Dev. 13, 239 252.
114. Verhagen, A. M., Ekert, P. G., Pakusch, M., Silke, J., Connolly, L. M.,
Reid, G. E., Moritz, R. L., Simpson, R. J., Vaux, D. L. (2000) Identifi-
cation of DIABLO, a mammalian protein that promotes apoptosis by
binding to and antagonizing IAP proteins. Cell 102, 4353.
115. Du, C., Fang, M., Li, Y., Li, L., Wang, X. (2000) Smac, a mitochondrial
protein that promotes cytochrome c-dependent caspase activation by
eliminating IAP inhibition. Cell 102, 33 42.
http://www.jleukbio.org
116. van den Broek, M. E., Kagi, D., Ossendorp, F., Toes, R., Vamvakas, S.,
Lutz, W. K., Melief, C. J., Zinkernagel, R. M., Hengartner, H. (1996)
Decreased tumor surveillance in perforin-deficient mice. J. Exp. Med.
184, 17811790.
117. Kagi, D., Ledermann, B., Burki, K., Seiler, P., Odermatt, B., Olsen, K. J.,
Podack, E. R., Zinkernagel, R. M., Hengartner, H. (1994) Cytotoxicity
mediated by T cells and natural killer cells is greatly impaired in
perforin-deficient mice. Nature 369, 3137.
118. Lowin, B., Beermann, F., Schmidt, A., Tschopp, J. (1994) A null muta-
tion in the perforin gene impairs cytolytic T lymphocyte- and natural
killer cell-mediated cytotoxicity. Proc. Natl. Acad. Sci. USA 91, 11571
11575.
119. Heusel, J. W., Wesselschmidt, R. L., Shresta, S., Russell, J. H., Ley, T. J.
(1994) Cytotoxic lymphocytes require granzyme B for the rapid induction
of DNA fragmentation and apoptosis in allogeneic target cells. Cell 76,
977987.
120. Sarin, A., Williams, M. S., Alexander-Miller, M. A., Berzofsky, J. A.,
Zacharchuk, C. M., Henkart, P. A. (1997) Target cell lysis by CTL
granule exocytosis is independent of ICE/Ced-3 family proteases. Immu-
nity 6, 209 215.
121. Medema, J. P., Toes, R. E., Scaffidi, C., Zheng, T. S., Flavell, R. A.,
Melief, C. J., Peter, M. E., Offringa, R., Krammer, P. H. (1997) Cleavage
of FLICE (caspase-8) by granzyme B during cytotoxic T lymphocyte-
induced apoptosis. Eur. J. Immunol. 27, 34923498.
122. Heibein, J. A., Goping, I. S., Barry, M., Pinkoski, M. J., Shore, G. C.,
Green, D. R., Bleackley, R. C. (2000) Granzyme B-mediated cytochrome
c release is regulated by the Bcl-2 family members Bid and Bax. J. Exp.
Med. 192, 13911402.
123. van den Broek, M. E., Kagi, D., Ossendorp, F., Toes, R., Vamvakas, S.,
Lutz, W. K., Melief, C. J., Zinkernagel, R. M., Hengartner, H. (1996)
Decreased tumor surveillance in perforin-deficient mice. J. Exp. Med.
184, 17811790.
124. Smyth, M. J., Thia, K. Y., Street, S. E., MacGregor, D., Godfrey, D. I.,
Trapani, J. A. (2000) Perforin-mediated cytotoxicity is critical for sur-
veillance of spontaneous lymphoma. J. Exp. Med. 192, 755760.
125. Mullbacher, A., Waring, P., Tha Hla, R., Tran, T., Chin, S., Stehle, T.,
Museteanu, C., Simon, M. M. (1999) Granzymes are the essential down-
stream effector molecules for the control of primary virus infections by
cytolytic leukocytes. Proc. Natl. Acad. Sci. USA 96, 13950 13955.
126. Davis, J. E., Smyth, M. J., Trapani, J. A. (2001) Granzyme A- and
B-deficient killer lymphocytes are defective in eliciting DNA fragmen-
tation but retain potent in vivo anti-tumor capacity. Eur. J. Immunol. 31,
39 47.
127. Witko-Sarsat, V., Rieu, P., Descamps-Latscha, B., Lesavre, P., Halb-
wachs-Mecarelli, L. (2000) Neutrophils: molecules, functions and patho-
physiological aspects. Lab. Investig. 80, 617 653.
128. Babior, B. M. (1999) NADPH oxidase: an update. Blood 93, 1464 1476.
129. Bastian, N. R., Hibbs Jr., J. B. (1994) Assembly and regulation of
NADPH oxidase and nitric oxide synthase. Curr. Opin. Immunol. 6,
131139.
130. Hampton, M. B., Kettle, A. J., Winterbourn, C. C. (1998) Inside the
neutrophil phagosome: oxidants, myeloperoxidase, and bacterial killing.
Blood 92, 30073017.
131. Bogdan, C., Rollinghoff, M., Diefenbach, A. (2000) The role of nitric
oxide in innate immunity. Immunol. Rev. 173, 1726.
132. Gudmundsson, G. H., Agerberth, B. (1999) Neutrophil antibacterial
peptides, multifunctional effector molecules in the mammalian immune
system. J. Immunol. Methods 232, 4554.
133. Shankaran, V., Ikeda, H., Bruce, A. T., White, J. M., Swanson, P. E., Old,
L. J., Schreiber, R. D. (2001) IFNgamma and lymphocytes prevent
primary tumour development and shape tumour immunogenicity. Nature
410, 11071111.
134. Davidson, W. F., Giese, T., Fredrickson, T. N. (1998) Spontaneous
development of plasmacytoid tumors in mice with defective Fas-Fas
ligand interactions. J. Exp. Med. 187, 18251838.
135. Irmler, M., Thome, M., Hahne, M., Schneider, P., Hofmann, K., Steiner,
V., Bodmer, J. L., Schroter, M., Burns, K., Mattmann, C., Rimoldi, D.,
French, L. E., Tschopp, J. (1997) Inhibition of death receptor signals by
cellular FLIP. Nature 388, 190 195.
136. Kataoka, T., Schroter, M., Hahne, M., Schneider, P., Irmler, M., Thome,
M., Froelich, C. J., Tschopp, J. (1998) FLIP prevents apoptosis induced
by death receptors but not by perforin/granzyme B, chemotherapeutic
drugs, and gamma irradiation. J. Immunol. 161, 3936 3942.
137. Medema, J. P., de Jong, J., van Hall, T., Melief, C. J., Offringa, R. (1999)
Immune escape of tumors in vivo by expression of cellular FLICE-
inhibitory protein. J. Exp. Med. 190, 10331038.
138. Taylor, M. A., Chaudhary, P. M., Klem, J., Kumar, V., Schatzle, J. D.,
Bennett, M. (2001) Inhibition of the death receptor pathway by cFLIP
confers partial engraftment of MHC class I-deficient stem cells and
reduces tumor clearance in perforin-deficient mice. J. Immunol. 167,
4230 4237.
139. Griffith, T. S., Chin, W. A., Jackson, G. C., Lynch, D. H., Kubin, M. Z.
(1998) Intracellular regulation of TRAIL-induced apoptosis in human
melanoma cells. J. Immunol. 161, 28332840.
140. Mueller, C. M., Scott, D. W. (2000) Distinct molecular mechanisms of
Fas resistance in murine B lymphoma cells. J. Immunol. 165, 1854
1862.
141. Tepper, C. G., Seldin, M. F. (1999) Modulation of caspase-8 and FLICE-
inhibitory protein expression as a potential mechanism of Epstein-Barr
virus tumorigenesis in Burkitts lymphoma. Blood 94, 17271737.
142. Thome, M., Schneider, P., Hofmann, K., Fickenscher, H., Meinl, E.,
Neipel, F., Mattmann, C., Burns, K., Bodmer, J. L., Schroter, M., Scaffidi,
C., Krammer, P. H., Peter, M. E., Tschopp, J. (1997) Viral FLICE-
inhibitory proteins (FLIPs) prevent apoptosis induced by death receptors.
Nature 386, 517521.
143. Hu, S., Vincenz, C., Ni, J., Gentz, R., Dixit, V. M. (1997) I-FLICE, a
novel inhibitor of tumor necrosis factor receptor-1- and CD-95-induced
apoptosis. J. Biol. Chem. 272, 1725517257.
144. Bertin, J., Armstrong, R. C., Ottilie, S., Martin, D. A., Wang, Y., Banks,
S., Wang, G. H., Senkevich, T. G., Alnemri, E. S., Moss, B., Lenardo,
M. J., Tomaselli, K. J., Cohen, J. I. (1997) Death effector domain-
containing herpesvirus and poxvirus proteins inhibit both Fas- and
TNFR1-induced apoptosis. Proc. Natl. Acad. Sci. USA 94, 11721176.
145. Djerbi, M., Screpanti, V., Catrina, A. I., Bogen, B., Biberfeld, P., Gran-
dien, A. (1999) The inhibitor of death receptor signaling, FLICE-inhib-
itory protein defines a new class of tumor progression factors. J. Exp.
Med. 190, 10251032.
146. Stamatopoulos, K., Kosmas, C., Belessi, C., Stavroyianni, N., Kyriazo-
poulos, P., Papadaki, T. (2000) Molecular insights into the immunopatho-
genesis of follicular lymphoma. Immunol. Today 21, 298 305.
147. Tsujimoto, Y., Cossman, J., Jaffe, E., Croce, C. M. (1985) Involvement of
the bcl-2 gene in human follicular lymphoma. Science 228, 1440 1443.
148. McDonnell, T. J., Deane, N., Platt, F. M., Nunez, G., Jaeger, U., Mc-
Kearn, J. P., Korsmeyer, S. J. (1989) bcl-2-immunoglobulin transgenic
mice demonstrate extended B cell survival and follicular lymphoprolif-
eration. Cell 57, 79 88.
149. Reed, J. C., Cuddy, M., Slabiak, T., Croce, C. M., Nowell, P. C. (1988)
Oncogenic potential of bcl-2 demonstrated by gene transfer. Nature 336,
259 261.
150. Strasser, A., Harris, A. W., Bath, M. L., Cory, S. (1990) Novel primitive
lymphoid tumours induced in transgenic mice by cooperation between
myc and bcl-2. Nature 348, 331333.
151. Vaux, D. L., Cory, S., Adams, J. M. (1988) Bcl-2 gene promotes haemo-
poietic cell survival and cooperates with c-myc to immortalize pre-B
cells. Nature 335, 440 442.
152. Fanidi, A., Harrington, E. A., Evan, G. I. (1992) Cooperative interaction
between c-myc and bcl-2 proto-oncogenes. Nature 359, 554 556.
153. Kogan, S. C., Brown, D. E., Shultz, D. B., Truong, B. T., Lallemand-
Breitenbach, V., Guillemin, M. C., Lagasse, E., Weissman, I. L., Bishop,
J. M. (2001) BCL-2 cooperates with promyelocytic leukemia retinoic acid
receptor alpha chimeric protein (PMLRARalpha) to block neutrophil
differentiation and initiate acute leukemia. J. Exp. Med. 193, 531543.
154. Weller, M., Malipiero, U., Aguzzi, A., Reed, J. C., Fontana, A. (1995)
Protooncogene bcl-2 gene transfer abrogates Fas/APO-1 antibody-medi-
ated apoptosis of human malignant glioma cells and confers resistance to
chemotherapeutic drugs and therapeutic irradiation. J. Clin. Investig. 95,
26332643.
155. Campos, L., Rouault, J. P., Sabido, O., Oriol, P., Roubi, N., Vasselon, C.,
Archimbaud, E., Magaud, J. P., Guyotat, D. (1993) High expression of
bcl-2 protein in acute myeloid leukemia cells is associated with poor
response to chemotherapy. Blood 81, 30913096.
156. Hermine, O., Haioun, C., Lepage, E., dAgay, M. F., Briere, J., Lavignac,
C., Fillet, G., Salles, G., Marolleau, J. P., Diebold, J., Reyas, F., Gaulard,
P. (1996) Prognostic significance of bcl-2 protein expression in aggres-
sive non-Hodgkins lymphoma. Groupe dEtude des Lymphomes de
lAdulte (GELA). Blood 87, 265272.
157. Miyashita, T., Reed, J. C. (1992) bcl-2 Gene transfer increases relative
resistance of S49.1 and WEHI7.2 lymphoid cells to cell death and DNA
fragmentation induced by glucocorticoids and multiple chemotherapeutic
drugs. Cancer Res. 52, 54075411.
158. Schmitt, C. A., Rosenthal, C. T., Lowe, S. W. (2000) Genetic analysis of
chemoresistance in primary murine lymphomas. Nat. Med. 6, 1029
1035.
159. Findley, H. W., Gu, L., Yeager, A. M., Zhou, M. (1997) Expression and
regulation of Bcl-2, Bcl-xl, and Bax correlate with p53 status and
Igney and Krammer Apoptosis and tumor immune escape 917
 sensitivity to apoptosis in childhood acute lymphoblastic leukemia.
Blood 89, 2986 2993.
160. Coustan-Smith, E., Kitanaka, A., Pui, C. H., McNinch, L., Evans, W. E.,
Raimondi, S. C., Behm, F. G., Arico, M., Campana, D. (1996) Clinical
relevance of BCL-2 overexpression in childhood acute lymphoblastic
leukemia. Blood 87, 1140 1146.
161. Henderson, S., Huen, D., Rowe, M., Dawson, C., Johnson, G., Rickinson,
A. (1993) Epstein-Barr virus-coded BHRF1 protein, a viral homologue of
Bcl-2, protects human B cells from programmed cell death. Proc. Natl.
Acad. Sci. USA 90, 8479 8483.
162. Pearson, G. R., Luka, J., Petti, L., Sample, J., Birkenbach, M., Braun, D.,
Kieff, E. (1987) Identification of an Epstein-Barr virus early gene en-
coding a second component of the restricted early antigen complex.
Virology 160, 151161.
163. Tarodi, B., Subramanian, T., Chinnadurai, G. (1994) Epstein-Barr virus
BHRF1 protein protects against cell death induced by DNA-damaging
agents and heterologous viral infection. Virology 201, 404 407.
164. Sarid, R., Sato, T., Bohenzky, R. A., Russo, J. J., Chang, Y. (1997)
Kaposis sarcoma-associated herpesvirus encodes a functional bcl-2 ho-
mologue. Nat. Med. 3, 293298.
165. Boise, L. H., Gonzalez-Garcia, M., Postema, C. E., Ding, L., Lindsten, T.,
Turka, L. A., Mao, X., Nunez, G., Thompson, C. B. (1993) bcl-x, a
bcl-2-Related gene that functions as a dominant regulator of apoptotic
cell death. Cell 74, 597 608.
166. Dole, M. G., Jasty, R., Cooper, M. J., Thompson, C. B., Nunez, G., Castle,
V. P. (1995) Bcl-xL is expressed in neuroblastoma cells and modulates
chemotherapy-induced apoptosis. Cancer Res. 55, 2576 2582.
167. Minn, A. J., Rudin, C. M., Boise, L. H., Thompson, C. B. (1995)
Expression of bcl-xL can confer a multidrug resistance phenotype. Blood
86, 19031910.
168. Zhou, P., Qian, L., Kozopas, K. M., Craig, R. W. (1997) Mcl-1, a Bcl-2
family member, delays the death of hematopoietic cells under a variety
of apoptosis-inducing conditions. Blood 89, 630 643.
169. Reed, J. C. (2001) The Survivin saga goes in vivo. J. Clin. Investig. 108,
965969.
170. Ambrosini, G., Adida, C., Altieri, D. C. (1997) A novel anti-apoptosis
gene, survivin, expressed in cancer and lymphoma. Nat. Med. 3, 917
921.
171. Adida, C., Berrebi, D., Peuchmaur, M., Reyes-Mugica, M., Altieri, D. C.
(1998) Anti-apoptosis gene, survivin, and prognosis of neuroblastoma.
Lancet 351, 882 883.
172. Grossman, D., Kim, P. J., Blanc-Brude, O. P., Brash, D. E., Tognin, S.,
Marchisio, P. C., Altieri, D. C. (2001) Transgenic expression of survivin
in keratinocytes counteracts UVB-induced apoptosis and cooperates with
loss of p53. J. Clin. Investig. 108, 991999.
173. Grossman, D., Kim, P. J., Schechner, J. S., Altieri, D. C. (2001) Inhibi-
tion of melanoma tumor growth in vivo by survivin targeting. Proc. Natl.
Acad. Sci. USA 98, 635 640.
174. Mesri, M., Wall, N. R., Li, J., Kim, R. W., Altieri, D. C. (2001) Cancer
gene therapy using a survivin mutant adenovirus. J. Clin. Investig. 108,
981990.
175. Dierlamm, J., Baens, M., Wlodarska, I., Stefanova-Ouzounova, M., Her-
nandez, J. M., Hossfeld, D. K., De Wolf-Peeters, C., Hagemeijer, A., Van
den Berghe, H., Marynen, P. (1999) The apoptosis inhibitor gene API2
and a novel 18q gene, MLT, are recurrently rearranged in the t(11;
18)(q21;q21)p6ssociated with mucosa-associated lymphoid tissue lym-
phomas. Blood 93, 36013609.
176. Bird, C. H., Sutton, V. R., Sun, J., Hirst, C. E., Novak, A., Kumar, S.,
Trapani, J. A., Bird, P. I. (1998) Selective regulation of apoptosis: the
cytotoxic lymphocyte serpin proteinase inhibitor 9 protects against gran-
zyme B-mediated apoptosis without perturbing the Fas cell death path-
way. Mol. Cell. Biol. 18, 6387 6398.
177. Medema, J. P., de Jong, J., Peltenburg, L. T., Verdegaal, E. M., Gorter,
A., Bres, S. A., Franken, K. L., Hahne, M., Albar, J. P., Melief, C. J.,
Offringa, R. (2001) Blockade of the granzyme B/perforin pathway through
overexpression of the serine protease inhibitor PI-9/SPI-6 constitutes a
mechanism for immune escape by tumors. Proc. Natl. Acad. Sci. USA 98,
1151511520.
178. Bladergroen, B. A., Meijer, C. J., ten Berge, R. L., Hack, C. E., Muris,
J. J., Dukers, D. F., Chott, A., Kazama, Y., Oudejans, J. J., van Berkum,
O., Kummer, J. A. (2002) Expression of the granzyme B inhibitor,
protease inhibitor 9, by tumor cells in patients with non-Hodgkin and
Hodgkin lymphoma: a novel protective mechanism for tumor cells to
circumvent the immune system? Blood 99, 232237.
179. Cheng, J., Zhou, T., Liu, C., Shapiro, J. P., Brauer, M. J., Kiefer, M. C.,
Barr, P. J., Mountz, J. D. (1994) Protection from Fas-mediated apoptosis
by a soluble form of the Fas molecule. Science 263, 1759 1762.
918 Journal of Leukocyte Biology Volume 71, June 2002
180. Midis, G. P., Shen, Y., Owen-Schaub, L. B. (1996) Elevated soluble Fas
(sFas) levels in nonhematopoietic human malignancy. Cancer Res. 56,
3870 3874.
181. Ugurel, S., Rappl, G., Tilgen, W., Reinhold, U. (2001) Increased soluble
CD95 (sFas/CD95) serum level correlates with poor prognosis in mela-
noma patients. Clin. Cancer Res. 7, 12821286.
182. Gerharz, C. D., Ramp, U., Dejosez, M., Mahotka, C., Czarnotta, B.,
Bretschneider, U., Lorenz, I., Muller, M., Krammer, P. H., Gabbert, H. E.
(1999) Resistance to CD95 (APO-1/Fas)-mediated apoptosis in human
renal cell carcinomas: an important factor for evasion from negative
growth control. Lab. Investig. 79, 15211534.
183. Bai, C., Connolly, B., Metzker, M. L., Hilliard, C. A., Liu, X., Sandig, V.,
Soderman, A., Galloway, S. M., Liu, Q., Austin, C. P., Caskey, C. T.
(2000) Overexpression of M68/DcR3 in human gastrointestinal tract
tumors independent of gene amplification and its location in a four-gene
cluster. Proc. Natl. Acad. Sci. USA 97, 1230 1235.
184. Roth, W., Isenmann, S., Nakamura, M., Platten, M., Wick, W., Kleihues,
P., Bahr, M., Ohgaki, H., Ashkenazi, A., Weller, M. (2001) Soluble decoy
receptor 3 is expressed by malignant gliomas and suppresses CD95
ligand-induced apoptosis and chemotaxis. Cancer Res. 61, 2759 2765.
185. Lehmann, C., Zeis, M., Schmitz, N., Uharek, L. (2000) Impaired binding
of perforin on the surface of tumor cells is a cause of target cell resistance
against cytotoxic effector cells. Blood 96, 594 600.
186. Strand, S., Hofmann, W. J., Hug, H., Muller, M., Otto, G., Strand, D.,
Mariani, S. M., Stremmel, W., Krammer, P. H., Galle, P. R. (1996)
Lymphocyte apoptosis induced by CD95 (APO-1/Fas) ligand-expressing
tumor cellsa mechanism of immune evasion? Nat. Med. 2, 13611366.
187. Higaki, K., Yano, H., Kojiro, M. (1996) Fas antigen expression and its
relationship with apoptosis in human hepatocellular carcinoma and
noncancerous tissues. Am. J. Pathol. 149, 429 437.
188. Moller, P., Koretz, K., Leithauser, F., Bruderlein, S., Henne, C., Quent-
meier, A., Krammer, P. H. (1994) Expression of APO-1 (CD95), a
member of the NGF/TNF receptor superfamily, in normal and neoplastic
colon epithelium. Int. J. Cancer 57, 371377.
189. Leithauser, F., Dhein, J., Mechtersheimer, G., Koretz, K., Bruderlein, S.,
Henne, C., Schmidt, A., Debatin, K. M., Krammer, P. H., Moller, P.
(1993) Constitutive and induced expression of APO-1, a new member of
the nerve growth factor/tumor necrosis factor receptor superfamily, in
normal and neoplastic cells. Lab. Investig. 69, 415 429.
190. Volkmann, M., Schiff, J. H., Hajjar, Y., Otto, G., Stilgenbauer, F., Fiehn,
W., Galle, P. R., Hofmann, W. J. (2001) Loss of CD95 expression is
linked to most but not all p53 mutants in European hepatocellular
carcinoma. J. Mol. Med. 79, 594 600.
191. Peli, J., Schroter, M., Rudaz, C., Hahne, M., Meyer, C., Reichmann, E.,
Tschopp, J. (1999) Oncogenic Ras inhibits Fas ligand-mediated apopto-
sis by downregulating the expression of Fas. EMBO J. 18, 1824 1831.
192. Maeda, T., Yamada, Y., Moriuchi, R., Sugahara, K., Tsuruda, K., Joh, T.,
Atogami, S., Tsukasaki, K., Tomonaga, M., Kamihira, S. (1999) Fas gene
mutation in the progression of adult T cell leukemia. J. Exp. Med. 189,
10631071.
193. Landowski, T. H., Qu, N., Buyuksal, I., Painter, J. S., Dalton, W. S.
(1997) Mutations in the Fas antigen in patients with multiple myeloma.
Blood 90, 4266 4270.
194. Cascino, I., Papoff, G., De Maria, R., Testi, R., Ruberti, G. (1996)
Fas/Apo-1 (CD95) receptor lacking the intracytoplasmic signaling do-
main protects tumor cells from Fas-mediated apoptosis. J. Immunol. 156,
1317.
195. Straus, S. E., Jaffe, E. S., Puck, J. M., Dale, J. K., Elkon, K. B.,
Rosen-Wolff, A., Peters, A. M., Sneller, M. C., Hallahan, C. W., Wang,
J., Fischer, R. E., Jackson, C. M., Lin, A. Y., Baumler, C., Siegert, E.,
Marx, A., Vaishnaw, A. K., Grodzicky, T., Fleisher, T. A., Lenardo, M. J.
(2001) The development of lymphomas in families with autoimmune
lymphoproliferative syndrome with germline Fas mutations and defective
lymphocyte apoptosis. Blood 98, 194 200.
196. Shin, M. S., Kim, H. S., Lee, S. H., Park, W. S., Kim, S. Y., Park, J. Y.,
Lee, J. H., Lee, S. K., Lee, S. N., Jung, S. S., Han, J. Y., Kim, H., Lee,
J. Y., Yoo, N. J. (2001) Mutations of tumor necrosis factor-related
apoptosis-inducing ligand receptor 1 (TRAIL-R1) and receptor 2
(TRAIL-R2) genes in metastatic breast cancers. Cancer Res. 61, 4942
4946.
197. Fisher, M. J., Virmani, A. K., Wu, L., Aplenc, R., Harper, J. C., Powell,
S. M., Rebbeck, T. R., Sidransky, D., Gazdar, A. F., El-Deiry, W. S.
(2001) Nucleotide substitution in the ectodomain of trail receptor DR4 is
associated with lung cancer and head and neck cancer. Clin. Cancer Res.
7, 1688 1697.
198. Pai, S. I., Wu, G. S., Ozoren, N., Wu, L., Jen, J., Sidransky, D., El-Deiry,
W. S. (1998) Rare loss-of-function mutation of a death receptor gene in
head and neck cancer. Cancer Res 58, 35133518.
http://www.jleukbio.org
199. Lee, S. H., Shin, M. S., Kim, H. S., Lee, H. K., Park, W. S., Kim, S. Y.,
Lee, J. H., Han, S. Y., Park, J. Y., Oh, R. R., Jang, J. J., Han, J. Y., Lee,
J. Y., Yoo, N. J. (1999) Alterations of the DR5/TRAIL receptor 2 gene in
non-small cell lung cancers. Cancer Res. 59, 56835686.
200. Teitz, T., Wei, T., Valentine, M. B., Vanin, E. F., Grenet, J., Valentine,
V. A., Behm, F. G., Look, A. T., Lahti, J. M., Kidd, V. J. (2000) Caspase
8 is deleted or silenced preferentially in childhood neuroblastomas with
amplification of MYCN. Nat. Med. 6, 529 535.
201. Rampino, N., Yamamoto, H., Ionov, Y., Li, Y., Sawai, H., Reed, J. C.,
Perucho, M. (1997) Somatic frameshift mutations in the BAX gene in
colon cancers of the microsatellite mutator phenotype. Science 275,
967969.
202. Meijerink, J. P., Mensink, E. J., Wang, K., Sedlak, T. W., Sloetjes, A. W.,
de Witte, T., Waksman, G., Korsmeyer, S. J. (1998) Hematopoietic
malignancies demonstrate loss-of-function mutations of BAX. Blood 91,
29912997.
203. Molenaar, J. J., Gerard, B., Chambon-Pautas, C., Cave, H., Duval, M.,
Vilmer, E., Grandchamp, B. (1998) Microsatellite instability and frame-
shift mutations in BAX and transforming growth factor-beta RII genes are
very uncommon in acute lymphoblastic leukemia in vivo but not in cell
lines. Blood 92, 230 233.
204. Krajewski, S., Blomqvist, C., Franssila, K., Krajewska, M., Wasenius,
V. M., Niskanen, E., Nordling, S., Reed, J. C. (1995) Reduced expression
of proapoptotic gene BAX is associated with poor response rates to
combination chemotherapy and shorter survival in women with meta-
static breast adenocarcinoma. Cancer Res. 55, 4471 4478.
205. Yin, C., Knudson, C. M., Korsmeyer, S. J., Van Dyke, T. (1997) Bax
suppresses tumorigenesis and stimulates apoptosis in vivo. Nature 385,
637 640.
206. Bargou, R. C., Wagener, C., Bommert, K., Mapara, M. Y., Daniel, P. T.,
Arnold, W., Dietel, M., Guski, H., Feller, A., Royer, H. D., Dorken, B.
(1996) Overexpression of the death-promoting gene bax-alpha which is
downregulated in breast cancer restores sensitivity to different apoptotic
stimuli and reduces tumor growth in SCID mice. J. Clin. Investig. 97,
26512659.
207. Ionov, Y., Yamamoto, H., Krajewski, S., Reed, J. C., Perucho, M. (2000)
Mutational inactivation of the proapoptotic gene BAX confers selective
advantage during tumor clonal evolution. Proc. Natl. Acad. Sci. USA 97,
1087210877.
208. Soengas, M. S., Capodieci, P., Polsky, D., Mora, J., Esteller, M., Opitz-
Araya, X., McCombie, R., Herman, J. G., Gerald, W. L., Lazebnik, Y. A.,
Cordon-Cardo, C., Lowe, S. W. (2001) Inactivation of the apoptosis
effector Apaf-1 in malignant melanoma. Nature 409, 207211.
209. Krammer, P. H. (1997) The tumor strikes back. Cell Death Differ. 4,
362364.
210. Igney, F. H., Behrens, C. K., Krammer, P. H. (2000) Tumor counterat-
tack concept and reality. Eur. J. Immunol. 30, 725731.
211. Walker, P. R., Saas, P., Dietrich, P. Y. (1998) Tumor expression of Fas
ligand (CD95L) and the consequences. Curr. Opin. Immunol. 10, 564
572.
212. French, L. E., Hahne, M., Viard, I., Radlgruber, G., Zanone, R., Becker,
K., Muller, C., Tschopp, J. (1996) Fas and Fas ligand in embryos and
adult mice: ligand expression in several immune-privileged tissues and
coexpression in adult tissues characterized by apoptotic cell turnover.
J. Cell Biol. 133, 335343.
213. Griffith, T. S., Brunner, T., Fletcher, S. M., Green, D. R., Ferguson, T. A.
(1995) Fas ligand-induced apoptosis as a mechanism of immune privi-
lege. Science 270, 1189 1192.
214. Stuart, P. M., Griffith, T. S., Usui, N., Pepose, J., Yu, X., Ferguson, T. A.
(1997) CD95 ligand (FasL)-induced apoptosis is necessary for corneal
allograft survival. J. Clin. Investig. 99, 396 402.
215. Griffith, T. S., Yu, X., Herndon, J. M., Green, D. R., Ferguson, T. A.
(1996) CD95-induced apoptosis of lymphocytes in an immune privileged
site induces immunological tolerance. Immunity 5, 716.
216. Bellgrau, D., Gold, D., Selawry, H., Moore, J., Franzusoff, A., Duke, R. C.
(1995) A role for CD95 ligand in preventing graft rejection. Nature 377,
630 632.
217. Allison, J., Georgiou, H. M., Strasser, A., Vaux, D. L. (1997) Transgenic
expression of CD95 ligand on islet beta cells induces a granulocytic
infiltration but does not confer immune privilege upon islet allografts.
Proc. Natl. Acad. Sci. USA 94, 39433947.
218. Saas, P., Walker, P. R., Hahne, M., Quiquerez, A. L., Schnuriger, V.,
Perrin, G., French, L., Van Meir, E. G., de Tribolet, N., Tschopp, J.,
Dietrich, P. Y. (1997) Fas ligand expression by astrocytoma in vivo:
maintaining immune privilege in the brain? J. Clin. Investig. 99, 1173
1178.
219. Choi, C., Park, J. Y., Lee, J., Lim, J. H., Shin, E. C., Ahn, Y. S., Kim,
C. H., Kim, S. J., Kim, J. D., Choi, I. S., Choi, I. H. (1999) Fas ligand and
Fas are expressed constitutively in human astrocytes and the expression
increases with IL-1, IL-6, TNF-alpha, or IFN-gamma. J. Immunol. 162,
1889 1895.
220. Lau, H. T., Yu, M., Fontana, A., Stoeckert Jr., C. J. (1996) Prevention of
islet allograft rejection with engineered myoblasts expressing FasL in
mice. Science 273, 109 112.
221. Kang, S. M., Hoffmann, A., Le, D., Springer, M. L., Stock, P. G., Blau,
H. M. (1997) Immune response and myoblasts that express Fas ligand.
Science 278, 13221324.
222. Strand, S., Galle, P. R. (1998) Immune evasion by tumours: involvement
of the CD95 (APO-1/Fas) system and its clinical implications. Mol. Med.
Today 4, 63 68.
223. Bennett, M. W., OConnell, J., OSullivan, G. C., Brady, C., Roche, D.,
Collins, J. K., Shanahan, F. (1998) The Fas counterattack in vivo:
apoptotic depletion of tumor-infiltrating lymphocytes associated with Fas
ligand expression by human esophageal carcinoma. J. Immunol. 160,
5669 5675.
224. Hahne, M., Rimoldi, D., Schroter, M., Romero, P., Schreier, M., French,
L. E., Schneider, P., Bornand, T., Fontana, A., Lienard, D., Cerottini, J.,
Tschopp, J. (1996) Melanoma cell expression of Fas(Apo-1/CD95) li-
gand: implications for tumor immune escape. Science 274, 13631366.
225. Mariani, S. M., Krammer, P. H. (1998) Differential regulation of TRAIL
and CD95 ligand in transformed cells of the T and B lymphocyte lineage.
Eur. J. Immunol. 28, 973982.
226. OConnell, J., OSullivan, G. C., Collins, J. K., Shanahan, F. (1996) The
Fas counterattack: Fas-mediated T cell killing by colon cancer cells
expressing Fas ligand. J. Exp. Med. 184, 10751082.
227. Villunger, A., Egle, A., Marschitz, I., Kos, M., Bock, G., Ludwig, H.,
Geley, S., Kofler, R., Greil, R. (1997) Constitutive expression of Fas
(Apo-1/CD95) ligand on multiple myeloma cells: a potential mechanism
of tumor-induced suppression of immune surveillance. Blood 90, 1220.
228. Nishimatsu, H., Takeuchi, T., Ueki, T., Kajiwara, T., Moriyama, N.,
Ishida, T., Li, B., Kakizoe, T., Kitamura, T. (1999) CD95 ligand expres-
sion enhances growth of murine renal cell carcinoma in vivo. Cancer
Immunol. Immunother. 48, 56 61.
229. Arai, H., Chan, S. Y., Bishop, D. K., Nabel, G. J. (1997) Inhibition of the
alloantibody response by CD95 ligand. Nat. Med. 3, 843 848.
230. Restifo, N. P. (2001) Countering the counterattack hypothesis. Nat.
Med. 7, 259.
231. Kang, S. M., Schneider, D. B., Lin, Z., Hanahan, D., Dichek, D. A.,
Stock, P. G., Baekkeskov, S. (1997) Fas ligand expression in islets of
Langerhans does not confer immune privilege and instead targets them
for rapid destruction. Nat. Med. 3, 738 743.
232. Allison, J., Georgiou, H. M., Strasser, A., Vaux, D. L. (1997) Transgenic
expression of CD95 ligand on islet beta cells induces a granulocytic
infiltration but does not confer immune privilege upon islet allografts.
Proc. Natl. Acad. Sci. USA 94, 39433947.
233. Judge, T. A., Desai, N. M., Yang, Z., Rostami, S., Alonso, L., Zhang, H.,
Chen, Y., Markman, J. F., DeMateo, R. P., Barker, C. F., Naji, A., Turka,
L. A. (1998) Utility of adenoviral-mediated Fas ligand gene transfer to
modulate islet allograft survival. Transplantation 66, 426 434.
234. Takeuchi, T., Ueki, T., Nishimatsu, H., Kajiwara, T., Ishida, T., Jishage,
K., Ueda, O., Suzuki, H., Li, B., Moriyama, N., Kitamura, T. (1999)
Accelerated rejection of Fas ligand-expressing heart grafts. J. Immunol.
162, 518 522.
235. Behrens, C. K., Igney, F. H., Arnold, B., Moller, P., Krammer, P. H.
(2001) CD95 ligand-expressing tumors are rejected in anti-tumor TCR
transgenic perforin knockout mice. J. Immunol. 166, 3240 3247.
236. Seino, K., Kayagaki, N., Okumura, K., Yagita, H. (1997) Antitumor effect
of locally produced CD95 ligand. Nat. Med. 3, 165170.
237. Shimizu, M., Fontana, A., Takeda, Y., Yagita, H., Yoshimoto, T., Mat-
suzawa, A. (1999) Induction of antitumor immunity with Fas/APO-1
ligand (CD95L)-transfected neuroblastoma neuro-2a cells. J. Immunol.
162, 7350 7357.
238. Okamoto, S., Takamizawa, S., Bishop, W., Wen, J., Kimura, K., Sandler,
A. (1999) Overexpression of Fas ligand does not confer immune privilege
to a pancreatic beta tumor cell line (betaTC-3). J. Surg. Res. 84, 77 81.
239. Yagita, H., Seino, K., Kayagaki, N., Okumura, K. (1996) CD95 ligand in
graft rejection. Nature 379, 682.
240. Arai, H., Gordon, D., Nabel, E. G., Nabel, G. J. (1997) Gene transfer of
Fas ligand induces tumor regression in vivo. Proc. Natl. Acad. Sci. USA
94, 1386213867.
241. Seino, K., Iwabuchi, K., Kayagaki, N., Miyata, R., Nagaoka, I., Matsu-
zawa, A., Fukao, K., Yagita, H., Okumura, K. (1998) Chemotactic
activity of soluble Fas ligand against phagocytes. J. Immunol. 161,
4484 4488.
Igney and Krammer Apoptosis and tumor immune escape 919
242. Ottonello, L., Tortolina, G., Amelotti, M., Dallegri, F. (1999) Soluble Fas
ligand is chemotactic for human neutrophilic polymorphonuclear leuko-
cytes. J. Immunol. 162, 36013606.
243. Shudo, K., Kinoshita, K., Imamura, R., Fan, H., Hasumoto, K., Tanaka,
M., Nagata, S., Suda, T. (2001) The membrane-bound but not the soluble
form of human Fas ligand is responsible for its inflammatory activity.
Eur. J. Immunol. 31, 2504 2511.
244. Hohlbaum, A. M., Moe, S., Marshak-Rothstein, A. (2000) Opposing
effects of transmembrane and soluble Fas ligand expression on inflam-
mation and tumor cell survival. J. Exp. Med. 191, 1209 1220.
245. Kang, S. M., Braat, D., Schneider, D. B., ORourke, R. W., Lin, Z.,
Ascher, N. L., Dichek, D. A., Baekkeskov, S., Stock, P. G. (2000) A
non-cleavable mutant of Fas ligand does not prevent neutrophilic de-
struction of islet transplants. Transplantation 69, 18131817.
246. Miwa, K., Asano, M., Horai, R., Iwakura, Y., Nagata, S., Suda, T. (1998)
Caspase 1-independent IL-1beta release and inflammation induced by
the apoptosis inducer Fas ligand. Nat. Med. 4, 12871292.
247. Hohlbaum, A. M., Gregory, M. S., Ju, S. T., Marshak-Rothstein, A.
(2001) Fas ligand engagement of resident peritoneal macrophages in vivo
induces apoptosis and the production of neutrophil chemotactic factors.
J. Immunol. 167, 6217 6224.
248. Rescigno, M., Piguet, V., Valzasina, B., Lens, S., Zubler, R., French, L.,
Kindler, V., Tschopp, J., Ricciardi-Castagnoli, P. (2000) Fas engagement
induces the maturation of dendritic cells (DCs), the release of interleukin
(IL)-1beta, and the production of interferon gamma in the absence of
IL-12 during DC-T cell cognate interaction: a new role for Fas ligand in
inflammatory responses. J. Exp. Med. 192, 16611668.
249. Muller, M., Strand, S., Hug, H., Heinemann, E. M., Walczak, H.,
Hofmann, W. J., Stremmel, W., Krammer, P. H., Galle, P. R. (1997)
Drug-induced apoptosis in hepatoma cells is mediated by the CD95
(APO-1/Fas) receptor/ligand system and involves activation of wild-type
p53. J. Clin. Investig. 99, 403 413.
250. Friesen, C., Herr, I., Krammer, P. H., Debatin, K. M. (1996) Involvement
of the CD95 (APO-1/FAS) receptor/ligand system in drug-induced apo-
ptosis in leukemia cells. Nat. Med. 2, 574 577.
920 Journal of Leukocyte Biology Volume 71, June 2002
251. Scaffidi, C., Kirchhoff, S., Krammer, P. H., Peter, M. E. (1999) Apoptosis
signaling in lymphocytes. Curr. Opin. Immunol. 11, 277285.
252. Klas, C., Debatin, K. M., Jonker, R. R., Krammer, P. H. (1993) Activa-
tion interferes with the APO-1 pathway in mature human T cells. Int.
Immunol. 5, 625 630.
253. Ehl, S., Hoffmann-Rohrer, U., Nagata, S., Hengartner, H., Zinkernagel,
R. (1996) Different susceptibility of cytotoxic T cells to CD95 (Fas/
Apo-1) ligand-mediated cell death after activation in vitro versus in vivo.
J. Immunol. 156, 23572360.
254. Chen, J. J., Sun, Y., Nabel, G. J. (1998) Regulation of the proinflamma-
tory effects of Fas ligand (CD95L). Science 282, 1714 1717.
255. Giovarelli, M., Musiani, P., Garotta, G., Ebner, R., Di Carlo, E., Kim, Y.,
Cappello, P., Rigamonti, L., Bernabei, P., Novelli, F., Modesti, A.,
Coletti, A., Ferrie, A. K., Lollini, P. L., Ruben, S., Salcedo, T., Forni, G.
(1999) A stealth effect: adenocarcinoma cells engineered to express
TRAIL elude tumor-specific and allogeneic T cell reactions. J. Immunol.
163, 4886 4893.
256. Mellado, M., de Ana, A. M., Moreno, M. C., Martinez, C., Rodriguez-
Frade, J. M. (2001) A potential immune escape mechanism by melanoma
cells through the activation of chemokine-induced T cell death. Curr.
Biol. 11, 691 696.
257. Keane, M. M., Ettenberg, S. A., Nau, M. M., Russell, E. K., Lipkowitz,
S. (1999) Chemotherapy augments TRAIL-induced apoptosis in breast
cell lines. Cancer Res. 59, 734 741.
258. Chinnaiyan, A. M., Prasad, U., Shankar, S., Hamstra, D. A., Shanaiah,
M., Chenevert, T. L., Ross, B. D., Rehemtulla, A. (2000) Combined effect
of tumor necrosis factor-related apoptosis-inducing ligand and ionizing
radiation in breast cancer therapy. Proc. Natl. Acad. Sci. USA 97,
1754 1759.
259. Jansen, B., Schlagbauer-Wadl, H., Brown, B. D., Bryan, R. N., van Elsas,
A., Muller, M., Wolff, K., Eichler, H. G., Pehamberger, H. (1998) bcl-2
Antisense therapy chemosensitizes human melanoma in SCID mice. Nat.
Med. 4, 232234.
260. Zamore, P. D. (2001) RNA interference: listening to the sound of silence.
Nat. Struct. Biol. 8, 746 750.
http://www.jleukbio.org