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 Previous Article | Table of Contents | Next Article 
Blood, Vol. 94 No. 12 (December 15), 1999:
pp. 4195-4201
Differential In Situ Cytokine Profiles of Langerhans-Like Cells and T
Cells in Langerhans Cell Histiocytosis: Abundant Expression of
Cytokines Relevant to Disease and Treatment
By
R. Maarten Egeler,
Blaise E. Favara,
Marjan van Meurs,
Jon D. Laman, and
Eric Claassen
From the Southern Alberta Children's Cancer Program, Alberta
Children's Hospital/Tom Baker Cancer Centre, Department of Oncology
and Pediatrics, University of Calgary, Calgary, Alberta, Canada; the
Department of Pediatric Oncology (Sophia Children's Hospital) and the
Department of Immunology, Erasmus University, Rotterdam, The
Netherlands; the Laboratory for Persistent Viral Diseases, Rocky
Mountain Laboratories, National Institutes of Health, Hamilton, MT; the
Department of Pathology, University of Utah, Salt Lake City, UT; and
the ID-DLO Institute for Animal Science and Health, Department of
Immunology, Lelystad, The Netherlands.
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ABSTRACT |
The pathogenesis of Langerhans cell histiocytosis (LCH) remains
poorly understood. To further elucidate LCH pathogenesis, we analyzed
the expression of 10 cytokines relevant to cellular recruitment and
activation at the protein level in 14 patients and identified the
lesional cells responsible for cytokine production in situ by
immunohistochemistry. The cytokines investigated included the
hematopoietic growth factors interleukin-3 (IL-3), IL-7, and granulocyte-macrophage colony-stimulating factor (GM-CSF); the lymphocyte regulatory cytokines IL-2, IL-4, and IL-10; the inflammatory regulators IL-1 and tumor necrosis factor- (TNF-); and the effector cell-activating cytokines IL-5 and interferon-
(IFN-). In all specimens, CD1a+ histiocytes
(LCH cells) and CD3+ T cells produced large amounts of
cytokines, creating a true cytokine storm. IL-2, IL-4, IL-5, and
TNF- were produced exclusively by T cells, whereas only IL-1 was
produced by LCH cells. Equal numbers of LCH cells, T cells, and
macrophages produced GM-CSF and IFN-. Equal numbers of LCH cells and
macrophages produced IL-10, whereas IL-3 was produced by T cells and
macrophages. IL-7 was only produced by macrophages. Eosinophils,
present in some specimens, were partially responsible for the
production of IL-5, IFN-, GM-CSF, IL-10, IL-3, and IL-7. Expression
of all cytokines, abundant in most biopsies, was irrespective of age,
gender, or site of biopsy. These findings emphasize the role of T cells
in LCH. The juxtaposition of T cells and LCH cells suggests that both
cells interact in a cytokine amplification cascade, resulting from
stimulation of autocrine and paracrine stimulatory loops. This cascade
can be linked directly to the development of LCH through recruitment,
maturation, and proliferation of LCH cells. The cytokines studied are
known to be involved in the development of other characteristic
features of LCH, such as fibrosis, necrosis, and osteolysis.
© 1999 by The American Society of Hematology.
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INTRODUCTION |
LANGERHANS CELL histiocytosis (LCH), a
rare disorder, mainly of children, is of unknown pathogenesis with
variable course.1 The broad clinical spectrum of disease
ranges from a single lytic lesion of bone that is usually cured by
simple curettage to a disseminated leukemia-like illness with
significant mortality. Intermediate forms of disease are chronic, often
with multiorgan involvement and diabetes insipidus.2
Lesions of LCH are polymorphous, featuring a monoclonal population of
CD1a+ histiocytes with a phenotype akin to that of cells of
the antigen-presenting Langerhans cell family. T cells, macrophages,
and eosinophils are variably present. The key histiocyte of LCH and its
normal counterpart, the Langerhans cell, express CD1a and S-100 and
contain Birbeck subcellular organelles.3 In contrast to
normal Langerhans cells, the principal histiocytes of LCH (LCH cells)
are actively proliferating, have a round rather than dendritic shape,
and express several contrasting antigenic markers.4
The morphology of LCH lesion and the clinical signs and symptoms of
disease suggest that cytokines may be important in the pathogenesis of
the disorder. Using immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR), upregulation of the
following cytokines in LCH lesions has previously been shown:
interleukin-1 (IL-1), IL-3, IL-4, IL-8, granulocyte-macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor-
(TNF-), transforming growth factor- (TGF-), and
leukemia inhibitory factor (LIF).5,6 These studies did not
identify the cellular source of these cytokines in the LCH lesion. We
have previously developed methods to analyze cytokine expression at the
protein level in situ so as to establish the identity of cells
producing an array of cytokines for both human and mouse
tissues.7,8 Using validated antibody-based
immunohistochemistry, intracellular cytokine present in
cytokine-producing cells can be reliably visualized, and
double-staining techniques for CD markers allow unequivocal identification of the cell type producing the cytokine. We used these
methods here to analyze the expression of an extended panel of
cytokines relevant to LCH pathogenesis and to identify their cellular
origin in 14 LCH lesions. We demonstrate that T cells and LCH cells are
the major local sources of cytokines, which are involved in recruitment
and survival of Langerhans cells, as well as in their maturation into
effector cells contributing to LCH pathogenesis.
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MATERIALS AND METHODS |
A definitive diagnosis of LCH and the representative nature of 14 specimens from 14 cases were confirmed by BEF. Eleven of the specimens
were from bone in cases of monoostotic LCH and 3 were from excisional
lymph node biopsies from patients with disseminated LCH. In all cases,
specimens were from initial diagnostic biopsies, with the duration of
symptoms and signs being less than 1 month before biopsy. All LCH
lesions were in the cellular phase of evolution.
Specimens were promptly snap-frozen and kept at  80°C until
sectioned and stained using an immuno-enzyme histochemical staining method.7,8 Briefly, frozen sections were cut at 8 µm in
an environment of 20°C. A series of 7 specimen sections
together with appropriate controls were placed on individual glass
slides and were kept overnight at room temperature in a closed
container with high humidity. After air-drying for 1 hour, sections
were fixed in acetone containing 0.01% hydrogen peroxide to block
endogenous peroxidase activity. Sections were then air-dried for 10 minutes and incubated with optimally diluted primary antibodies
(Table 1) overnight at 4°C in the dark.
Sections were washed twice in phosphate-buffered saline (PBS), and
second- and third-step incubations of 2 hours per RT each were
performed. After a PBS wash, horseradish peroxidase (HRP) activity was
shown with 3-aminocarbazole (AEC; Sigma, St Louis, MO) in
bright primary red.7,8 Alkaline phosphate was shown with
naphtol-AS-MX-phosphate (Sigma) and fast blue (BB base; Sigma) as
brilliant blue.7,8 Antibody 1/34 (DAKO Corp, Carpinteria,
CA) was used for CD1a and CD3 DAKO-EPOS, a rabbit polyclonal antibody coupled to an inert polymer backbone and HRP, was
used for CD3. Double stainings were performed, titrated, and evaluated
(red/blue and violet colors) as described before.7,8
The specificity of antibodies was determined as noted
earlier,8 but isotype-matched control antibodies were
always included (Fig 1A and B) to assess
nonspecific background staining (eg, by binding to Fc receptors). When
positive control stains were weak or negative, study results were
discarded. Negative controls uniformly showed no staining. Positive
control stainings on sections of human tonsil, spleen, and skin that
featured an allergic reaction and negative control stains on human
liver and skin were performed with each batch of study material on the
same individual slide (ie, on all slides). All staining was performed
in duplicate on separate slides on the same day and duplicate stains
were performed on different days. As in earlier studies,8
cells staining positive for cytokines were scored only when the
cytoplasm was brightly colored and the nucleus was clear and
transparent. When stains could not be reproduced because of lack of
tissue or lack of tissue integrity and/or lack of clearly discernible
cytology, they were not included in the data and were marked as not
determinate (Table 2). Two observers
(M.v.M. and E.C.) scored all slides independently and reconciled
differences in scoring by studying the slide(s) together. Slides were
scored for estimates of the number of positively staining lesional
cells. The frequency of cytokine expression/staining is indicated in
Table 2 as +++ reaction, meaning that the majority of cells in the
specimen produces that certain cytokine. This high level of expression
is illustrated in Fig 1C and D. Double staining for cell type and
cytokine was performed in at least 7 specimens for each cytokine
profile. We defined LCH cells as being CD1a+ and T cells as
CD3+. Eosinophils were identified by morphology, and
CD1a cells with morphological features of
histiocytes were called macrophages.
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| Fig 1.
In situ cytokine expression in LCH biopsies. Single
immunohistochemical labeling was used to assess cytokine production in
frozen sections of LCH biopsies. (A) IL-1 demonstrated in a positive
control tissue sample from human tonsil. Such control tissue was
included on each glass slide to internally validate different cytokine
stainings (original magnification × 100). (B) Human tonsil: negative
control staining using an isotype-matched primary antibody of
irrelevant specificity to exclude nonspecific binding of secondary and
tertiary step reagents (original magnification × 100).8
(C) IL-1-producing cells in a LCH lesion in bone (patient no.
S926704). Note the high density of cytokine-expressing cells and the
very intense staining of most of the cytoplasm, compared with (B)
(original magnification × 100). (D) IL-3-producing cells in a
monoostotic lesion (patient no. S9411100; original magnification × 100). (E) IL-10-producing cells in a affected lymph node in
disseminated LCH (patient no. S93292). Note that no clustering occurs
and that isolated cells display only partially cytokine-filled
cytoplasm (original magnification × 100). (F) GM-CSF-producing cells
in a monoostotic lesion (patient no. 9408P025; original magnification × 50).
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RESULTS |
The cytokine profile of the 14 LCH specimens is shown in Table 2. Some
cytokines were expressed so abundantly that staining resulted in the
blending of the images of individual cells. With the exception of IL-7,
IL-10, and GM-CSF, which were expressed by relatively few cells in most
specimens, all cytokines were consistently expressed at high levels.
The lowest levels of cytokine expression were seen in an isolated skull
lesion from an 11-year-old boy and a lymph node from a 2-year-old boy
with disseminated LCH. However, the cytokine profile on the latter was
limited. Eosinophils were obvious in 8 of the 14 specimens (Table 2),
and they were associated with expression of IL-3, IL-5, IL-7, IL-10,
interferon- (IFN-), and GM-CSF.
Under physiological conditions, IL-3 can stimulate the generation and
differentiation of progenitor cells of every lineage derived from the
pluripotential hematopoietic stem cells, including macrophages,
neutrophils, eosinophils, basophils, mast cells, megakaryocytes, and
erythroid cells. There was abundant expression of IL-3 in 6 of the 11 LCH specimens (Fig 1D), with 1 demonstrating no staining. Half of the
IL-3-producing cells were T cells and half were macrophages. In 5 specimens, IL-3 was also expressed by eosinophils. No IL-3 was produced
by LCH cells.
IL-7 is normally a major growth and differentiation factor for T cells
and B cells. It promotes immune effector functions in T
cells, natural killer (NK) cells, and monocytes-macrophages. There
was less IL-7 expression than any other cytokine in the 12 specimens
studied, and none was demonstrated in 4 specimens. The source of IL-7
in these lesions appeared to be macrophages. None was associated with T
cells or LCH cells. Eosinophils produced IL-7 in the 4 specimens.
The main activities of GM-CSF include the promotion of differentiation
and proliferation of hematologic progenitors. Modest numbers of cells
expressed GM-CSF (Fig 1F) in all but 1 (no expression) of the 13 specimens studied. All types of lesional cells, including eosinophils
in 4 specimens, produced GM-CSF.
IL-2 is the T-cell cytokine that stimulates ongoing specific immune
responses, T-cell differentiation, and synthesis of IFN-. IL-2 is
almost exclusively produced by T cells, but perhaps also by B cells.
IL-2, the most abundantly expressed cytokine in these specimens, was
absent in 1 specimen and was uniformly associated with the marker for T cells.
IL-4 can act on many cell types and at various stages of maturation as
it induces B cells to proliferate and to secrete IgG1. It also
induces T-cell proliferation. IL-4 modulates cyto- kine production by B cells, T cells, NK cells, monocytes-macrophages, and
several other cells and, therefore, plays an important role. Generally
IL-4 is very difficult to demonstrate in mouse and human tissue,
possibly due to low levels of expres- sion and/or to the use of
low-affinity antibodies. In contrast, staining for IL-4, which was
uniformly associated with T cells, was seen in all but 1 of the 8 LCH
specimens studied (Fig 2D).
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| Fig 2.
Identification of cellular origin of cytokines in LCH
biopsies. Double immunohistochemical labeling was used to determine
cytokine profiles of T cells and LCH cells in frozen sections of LCH
biopsies. IL-1 (red) is not produced by CD3+ T cells
(blue), as shown by the absence of intermediate (violet) staining
(original magnification × 100). (B) In contrast to (A), all
CD1a+ LCH cells (blue) also stain for IL-1 (red),
resulting in violet double staining (original magnification × 100).
(C) IL-5 (red) is not coexpressed with CD1a (blue) and is therefore not
produced by LCH cells (original magnification × 100). (D) IL-4
(red)-producing cells stain violet with anti-CD3 (blue) and hence are
T cells (original magnification × 100). (E) IFN--producing cells
stain in part double (note red, blue, and violet) with CD1a. Because
the same was shown in double stainings with anti-CD3, this cytokine is
produced by both T cells and LCH cells (original magnification × 200). (F) TNF--producing cells coexpress CD3 and are therefore
identified as T cells (original magnification × 100).
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IL-10 exhibits important functions in immune regulation, particularly
in controlling inflammatory responses via inhibition of
monocyte-macrophage activation. Very important as well are the potent
deactivating effects of IL-10 on monocytes-macrophages, granulocytes,
and dendritic cells. Modest expression of this cytokine was seen in 9 of 11 specimens (Fig 1E), and no staining was seen in 2. Staining was
associated equally with LCH cells and macrophages. Eosinophils also
expressed IL-10 in 3 specimens.
Cytokine IL-1 is a highly inflammatory cytokine and affects nearly
every cell type, often in concert with other cytokines. There was
expression by large numbers of cells in 8 of 14 specimens, with LCH
cells appearing to be the singular source of the cytokine (Fig 2A and B).
TNF- is produced during immune and host defense responses as a
primary mediator of immune regulation and the inflammatory response. In
9 of the 13 specimens tested, large numbers of T cells, the sole
source, expressed TNF- (Fig 2F).
Cytokine IL-5 is the main controling cytokine for eosinophilia. Only T
cells and eosinophils in 3 specimens produced IL-5 (Fig 2C).
IFN- is, in addition to its antiviral activities, an important
modulator of the immune system because of its ability to activate T
cells. Six specimens showed prominent expression with
equal contributions by LCH cells and T cells (Fig 2E). Faint expression was associated with macrophages in a few specimens, and eosinophils produced IFN- in 1.
There was a paucity of multinucleated giant histiocytes in the sections
from these specimens and, thus, too few were observed to establish the
cytokine profile of these lesional elements.
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DISCUSSION |
In this study, we demonstrate high expression of a large panel of
cytokines in LCH lesions at the protein level and, for the first time,
we show that T cells and LCH cells are the predominant sources of this
cytokine storm. LCH cells were the only source of IL-1 and a major
source of IL-10. Most of the cytokines were of T-cell origin,
suggesting a prominent role for this cell in the disease. The pattern
of cytokine expression forming this cytokine storm favors recruitment
of Langerhans cell progenitors as well as their maturation and rescue
from apoptosis, thereby explaining the pathologic accumulation of LCH
cells. The large number of cytokine-producing cells in LCH lesions
supports local amplification cascades of cellular proliferation and
activation, involving autocrine and paracrine stimulatory loops.
Several of the cytokines produced in LCH lesions directly contribute to
pathological sequelae of LCH, including fibrosis, bone resorption, and
necrosis. These findings are interpreted in an integrated view of the
pathology of LCH and contrasted with physiological Langerhans cell reactivity.
Langerhans Cells
The diagnostic lesional cell in LCH is most akin to the normal
Langerhans cell, with comparative data having been summarized recently.9 These cells, and only they, are monoclonal in
LCH lesions.10 The Langerhans cell, an immune cell of bone
marrow origin, is the sentinel of the human skin with respect to
foreign antigens. Strategically located in the epidermis, Langerhans
cells capture antigens upon entry, a process that is facilitated by their dendritic morphology. Antigen uptake, processing, and expression of antigenic peptides on the cell surface in the context of major histocompatibility complex (MHC)/HLA molecules leads to
differentiation and activation, including increased longevity of
MHC-peptide complexes at the cell surface.11 Langerhans
cells carrying antigen migrate through the lymph as veiled cells and
present antigen to peptide-specific T cells in the T-cell areas of
draining lymph nodes and spleen, where they are called interdigitating
dendritic cells.12 The interaction between Langerhans cells
and T cells needs to be bidirectional, because MHC plus antigenic
peptide, adhesion molecules, costimulatory molecules (eg, CD40-CD40L,
CD80/CD86), and cytokines are all required for complete activation of
both the antigen-presenting cell and the T cell.13
Langerhans Cells and Cytokines
The full range of cytokines produced by Langerhans cells and their
function needs to be further elucidated, but reviews on the
characteristics of Langerhans cells14 indicate that GM-CSF, IL-1, and TNF- are important in the development and trafficking of
Langerhans cells. Furthermore, TNF-, GM-CSF, and IL-3 enhance proliferation of Langerhans cells generated from CD34+ cord
blood or bone marrow cells.15 These in vitro-generated Langerhans cells, as well as the dendritic cells generated from peripheral blood, produce IL-1 and, when stimulated through CD40, also
secrete TNF-.16 Dermal Langerhans cell evolution to
lymph node interdigitating dendritic cells is enhanced by GM-CSF and IL-1.17,18 GM-CSF has a role in the recruitment of
Langerhans cells to various tissue sites.19 Expression of
relevant cytokine receptors on Langerhans cells has been confirmed,
including those for GM-CSF, IL-1, TNF-, and IFN- for
humans,20 whereas IL-2R is expressed by murine
cells21 and can be induced by CD40 ligation.16
Langerhans Cells Versus LCH Cells
LCH cells probably originate from the bone marrow and are very
sensitive to local factors such as cytokines and growth factors for
replication, maturation, and differentiation, as well as functional antigen processing and presentation. The issue as to which of the many
immunologically important cytokines are expressed in LCH lesions
remains a somewhat vexed one, because a limited number of biopsies have
been studied. Furthermore, in situ analysis is technically challenging,
and in vitro studies of both LCH cells and Langerhans cells are
confounded by contaminating cell populations producing cytokines.
Therefore, the goal in studying the production of cytokines in LCH is
to improve our understanding of disease pathogenesis and ultimately to
explore the potential of novel therapeutics such as recombinant
cytokines and antagonist molecules targeting costimulatory events. To
date, most studies on cytokine production in LCH lesions have focused
on descriptive analysis often based on mRNA detection and ignoring
which cells produce the different cytokines. We therefore analyzed
cytokine expression in LCH lesions at the protein level in a large
series of biopsies, including some cytokines studied in LCH previously,
but expanding the panel to a total of 10, with additional relevant
cytokines not evaluated before. Biopsy material of LCH is extremely
limited, so only confining studies are possible. This limited factor
forced us to use a restrictive panel of markers for the double staining studies. However, the double labeling in situ with CD3 for T cells and
CD1a for LCH cells enabled us to provide new insights into the identity
of cytokine-producing cells as well.
Pathogenic Roles of Cytokines in LCH Lesions
Indirect evidence for cytokine involvement in LCH.
Indirect evidence of a role for cytokines in LCH lies in the finding
that peripheral blood T-helper:suppressor ratios are high.22 This finding may be related to early studies
establishing that major Ig isotypes are elevated in 75% of patients
with LCH,23 because Th-cell cytokines are differential Ig
isotype switch factors and also stimulate B-cell proliferation and
antibody production. The positive clinical effects of
cyclosporin,24 thymic hormone,22 and bone
marrow transplantation25 further attest to a prominent role
of T cells and cytokines in LCH. Prostaglandin E2 and IL-1 have been
found in unseparated cell preparations of LCH lesions in vitro. IFN-
increases IL-1 secretion by LCH cells in vitro, indicating a role of T
cells in regulating the production of IL-1 in bone
lesions.26 T cells may stimulate Langerhans cells by IL-2
production, as evidenced by IL-2 receptor expression on murine epidermal Langerhans cells. In our study (Table 2), T cells
and LCH cells in most specimens produced IFN-, but in less than half did large numbers of cells stain for this cytokine. However, more cells
producing IFN- were seen in bone lesions than in lesions of lymph nodes.
Recruitment and differentiation of cells.
Previously, lesions of LCH have been shown to contain mRNA transcripts
for IL-1, IL-3, IL-4, IL-8, GM-CSF, TNF-, TGF-, and LIF. No
transcripts for IL-2, IL-5, IL-6, or IFN- were found.6 Using immunohistochemistry, De Graaf et al5
showed that cells in LCH lesions and those from normal and allergic
skin expressed IL-1, IL-1, GM-CSF, TGF-, TGF-, TNF-, and
IFN-, whereas LCH cells alone did not express basic fibroblast
growth factor. The cytokine IL-2 was not studied.5 Our data
complete these findings in showing that crucial immunologic active
cytokines, such as IL-2, IL-5, and IFN-, are, indeed, produced by
lesional T cells and that LCH cells produce IFN-, but they do not
produce IL-2 and IL-5. Because under physiological conditions IL-3 can stimulate the generation and differentiation of progenitor cells of
every lineage derived from the pluripotential hematopoietic stem cells,
IL-3 might contribute to the differentiation of LCH cell progenitors.
The distribution of LCH cells in the affected patient may be influenced
by GM-CSF27 (Table 2) and by adhesion molecules defined by
De Graaf et al.28 There are few studies of cytokine receptors in LCH, but Emile et al29 demonstrated the
expression of the GM-CSF receptor on LCH cells. Under physiological
conditions, GM-CSF increases differentiation and proliferation of
hematologic progenitors. Kaplan et al19 have shown that
GM-CSF is important in recruitment of Langerhans cells into different
tissues. Likewise, in LCH lesions, GM-CSF probably promotes growth and
differentiation of LCH cell progenitors. Elevated levels of GM-CSF were
found in the sera of patients with disseminated disease, but not in that of patients with localized LCH, suggesting that serum GM-CSF may
be a marker of the extent of the disease in LCH.30 Serum levels of IL-2 receptor show similar correlations.31
GM-CSF, IL-3, TNF-, and a number of other cytokines function as
chemoattractants for eosinophils, neutrophils, macrophages, and
CD34+ Langerhans cell precursors.32
Importantly, TNF- has been shown to inhibit spontaneous apoptosis of
Langerhans cells, promoting their survival,33,34,35 which
would explain the accumulation of LCH cells in lesions. Furthermore,
IL-1 promotes this effect of TNF-,33 as do GM-CSF and
IL-3,36 both of which we demonstrated in the lesions.
Accumulation of LCH cells may be further enhanced by stimulation of
Langerhans cell formation from their CD34+ progenitors by
GM-CSF and TNF-.36
Osteolysis and fibrosis.
IL-1 and TNF-, both amply expressed in LCH lesions (Table 2), and
IL-1 may synergistically enhance osteoclastic activity with
resultant osteolysis, the hallmark of the LCH lesion of bone. The
evolution of fibrosis in LCH lesions of bone, liver, and
lungs4 may be related to the lesional production of TGF-
(not studied here), which has been implicated as a potent sclerosing
agent.37 TNF- can recruit leukocytes and promote
angiogenesis as well as fibroblast proliferation in wound healing, and
similar activities might occur in the LCH lesions. Sclerosis has also
been implicated in the evolution of diabetes insipidus, the most common
endocrinopathy in LCH, secondary to obstructive involvement of the
hypothalamic/pituitary axis.
Rationale for 2-Chlorodeoxyadenosine (2-Cda) Treatment in LCH
This evidence of cross-talk between the LCH cell and the T cell in LCH
lesions raises prospects of a better understanding of the mechanism of
action of promising therapeutic agents such as 2-Cda in this
disorder.38,39 2-Cda is a purine analogue resistant to
degradation by adenosine deaminase. Phosphorylated derivates of 2-Cda
accumulate in cells with high deoxycytidine kinase activity, such as
lymphocytes, inducing changes in lymphocyte viability, resulting in DNA
strand breaks. The DNA damage is followed by a progressive decrease in
lymphocyte RNA synthesis, resulting in cell death.40 The
cytotoxic properties of 2-Cda are independent of cell division, making
it a powerful agent in the treatment of lymphoid neoplasms with
low-growth fractions, such as T-cell lymphomas.41 In vitro
and in vivo human monocytes are as sensitive as lymphocytes to these
effects of 2-Cda, following the same pathway.42 Until now,
the rationale for using 2-Cda in LCH was based on this latter
finding.38,39 Because Langerhans cells and
monocytes/macrophages are thought to originate from the same stem cell,
it was hypothesized that, because 2-Cda is toxic to monocytes, it might
be toxic to LCH cells. To confirm this, in vitro and in vivo studies
concerning the sensitivity of Langerhans cells or, even better, LCH
cells, should be undertaken. Until these studies are performed, the
results presented here showing that the LCH lesion is the result of
cross-talk between the LCH-cell and the activated T cell form a
rationale for using 2-Cda in LCH.
Perspective
The fact that clonality has been found in all LCH lesions reported to
date argues that LCH is a neoplastic disorder with varied biological
behavior, which could result from a genetic defect, an abnormal
response to infection, or an autoimmune phenomenon, but the true
significance of this clonality remains to be clarified. We have
demonstrated that a cytokine storm consisting of at least 10 major
cytokines occurs in LCH lesions. CD3+ T cells are pivotal,
because they produce 7 of 10 analyzed cytokines, whereas LCH cells
produce 4 of 10. We conclude that T cells are an important driving
force for the accumulation, proliferation, and differentiation of cells
in LCH lesions. The observed juxtaposition of T cells and LCH cells in
all tissues analyzed suggests intimate, if not cognate, interactions of
these 2 cell types, contributing to bidirectional stimulation and
cytokine production. Associated pathogenic effects of these cytokines
are likely to include chemotaxis of additional inflammatory cells,
overexpression of adhesion molecules, and fibrosis, necrosis, and
osteolysis. Systemic manifestations of LCH, such as macrophage
activation and fever, may reflect more far-reaching effects of these
cytokines. The current findings should prove useful in further rational
development of experimental LCH therapy.
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ACKNOWLEDGMENT |
The authors thank Louis Ribbens (TNO-Prevention and Health, Leiden, The
Netherlands) for performing the initial immunohistochemical analyses.
Some of the specimens were provided through the Pediatric Division of
the Cooperative Human Tissue Network, Children's Hospital (Columbus, OH).
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FOOTNOTES |
Submitted April 28, 1999; accepted August 3, 1999.
These studies were initiated through the Nikolas Symposium (organizers
Paul and Elizabeth Kontoyannis). The Histiocytosis Association of
America and the Histiocytosis Stichting in The Netherlands provided
additional financial support.
The publication costs of this
article were defrayed in part by
page charge payment. This article
must therefore be hereby marked
"advertisement"
in accordance with 18 U.S.C. section
1734 solely to indicate this fact.
Address reprint requests to R. Maarten Egeler, MD, PhD, Southern
Alberta Children's Cancer Program, Alberta Children's Hospital, 1820 Richmond Rd SW, Calgary, Alberta, Canada T2T 5C7; e-mail:
Maarten.Egeler{at}CRHA-Health.ab.ca.
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TOP
ABSTRACT
INTRODUCTION