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NEOPLASIA
From the Department of Research and Development,
Research Group on Lymphoma, EFS Rhône-Alpes Grenoble, La Tronche,
France; Schering Plough Laboratory for Immunological Research,
Dardilly, France; Groupe d'Etudes Immunologique des Leucémies,
Immunology Laboratory, Medicine Faculty, Vandoeuvre Les Nancy, France;
Clinical Hematological Department, Research Group on Lymphoma,
Michallon Hospital, Grenoble, France; Hematological Department, Dijon
Hospital, Dijon, France; Cellular Hematological Department, Dupuytren
Hospital, Limoges, France; Oncology Department, Bourg en Bresse
Hospital, Bourg en Bresse, France; Hematological Department, Annecy
Hospital, Annecy, France; and Hematological Department, Avicenne
Hospital, Bobigny, France.
This work aims to demonstrate that
CD4+CD56+ malignancies arise from transformed
cells of the lymphoid-related plasmacytoid dendritic cell (pDC) subset.
The analysis of malignant cells from 7 patients shows that in all
cases, like pDCs, leukemic cells are negative for lineage markers CD3,
CD19, CD13, CD33, and CD11c but express high levels of interleukin-3
receptor Several reports in the literature1-7
describe unusual and rare hematopoietic tumors expressing CD4 and CD56,
without CD3, CD19, CD13, and CD33 conventional lineage markers, and no
T- or B-cell receptor gene rearrangement. Clinically, these cases are characterized by a rapid aggressive course, an extranodal and notable
skin involvement, and a frequent evolution toward an overt leukemia.
The possible existence of a new entity has been raised, but the origin
of the tumor cells has not yet been resolved, even if they have
occasionally been classified as T,3 natural killer (NK),3-5 or myeloid5 precursors. In fact,
after comparison of their phenotype with that of all known
hematopoietic cells, we postulated that these cells are related to the
newly characterized plasmacytoid dendritic cell
(pDC).8
The pDCs are members of the heterogeneous dendritic cell family that
could be derived from human bone marrow9 and
thymus10,11 lymphoid-restricted progenitors. They have been
identified in peripheral blood12-14 and in T-cell areas of
tonsils,8 lymph nodes,15 and
thymus.16,17 The pDCs not only have a typical morphology,8,12 but they also have characteristic
phenotypic features such as expression of CD4, HLA-DR, and CD45RA while
they are lacking myeloid-related antigens CD11c, CD13, and
CD33.8,15,18 They are devoid of lineage-associated markers
such as CD3, CD14, CD19, and CD56, but CD2, CD5, and CD7 have
occasionally been observed.15,16 Noteworthy, they express
a low level of granulocyte-macrophage colony-stimulating factor
receptor In the present work, we identify CD4+, CD56+,
CD3 Patients and isolation of tumor cells
Effect of cytokines on cell viability
Differentiation of tumor cells into mature DCs Purified tumor cells from 5 patients were cultured at 106/mL for 3 or 6 days in complete 10% fetal calf serum medium supplemented or not with IL-3 (10 ng/mL). In 2 cases, maturation with CD40 ligation was analyzed: Irradiated (70 Gy) CD40L-transfected L cells (gift from P. Garonne, Schering-Plough) were added on day 4 to the IL-3 tumor cell culture (CD40L cells: tumor cell ratio = 1:5). On day 6, phenotypic and functional analyses were performed. Culture supernatants were cryopreserved for cytokine measurements.Flow cytometry and immunofluorescence Immunophenotype was analyzed by flow cytometry, using direct or indirect labeling, with the antibodies listed in Table 1. For comparison of expression levels, the mean fluorescence intensity ratio between specific mAb and isotypic control was calculated. Intracytoplasmic flow cytometric staining of perforin and granzyme B was performed after paraformaldehyde fixation (2%) and permeabilization of the cells (saponin 0.3%). Cytospins of cell suspensions were dehydrated, fixed with acetone ( 20°C, 10 minutes), and cryopreserved. After thawing and rehydratation, the
incubation with primary antibodies (perforin or CD83) was followed by
incubation with fluorescein isothiocyanate (FITC)-goat antimouse
immunoglobulin (Beckman Coulter Immunotech) and counterstaining with
Evan's blue dye (Sigma, Saint Quentin Fallavier, France).
Reverse transcriptase-polymerase chain reaction RNA was isolated from FACS-sorted normal blood pDCs, from tumor cells purified from 4 patients, from blood NK cells, and from monocyte-derived DCs obtained after a 6-day culture in GM-CSF and IL-13 (Sanofi, Labège, France) (mo-DCs) using MasterPure RNA purification kit (Epicentre Technologies, Madison, WI) according to the manufacturer's instructions. First-strand complementary DNA was prepared using oligo(dT) primers (Pharmacia, Uppsala, Sweden) and Superscript RNase-H reverse transcriptase (Gibco BRL, Gaithersburg, MD). Polymerase chain reaction (PCR) was performed in a DNA thermal cycler (PE Applied Biosystems, Foster City, CA) for 35 cycles (1 minute of denaturation at 94°C, 1 minute of annealing at 55°C for lambda-like 14.1 and granzyme B, or 60°C for IL-3R , FasL,
pre-T, perforin, CD56, and 2 minutes of elongation at 72°C) with
ampliTaq enzyme and buffer (Gene Amp PCR reagents kit, PE Applied
Biosystems, Foster City, CA), dNTPS at 10 mM (PE Applied Biosystems,
Foster City, CA), and dimethyl sulfoxide at 5% final concentration. The  -actin mRNA amplification was performed for 28 cycles at 60°C on the complementary DNA as positive control of
reaction efficiency. The primers used were as follows: IL-3R (sense:
5'-ATGCCGACTATTCTATGCCG-3', antisense: 5'-TGTCTCTGACCTGTTCTGTG-3'); lambda-like 14.1 (sense: 5'-ATGCATGCGGCCGCGGCATGTGTTTGGCAGC-3', antisense: 5'-ATCCGCGGCCGCATCGATAGGTCACCGTCA-AGATT-3'); pre-T (sense: 5'-GGCACACCCTTTCCTTCTCTG-3', antisense:
5'-GCAGGTCCTGGCTGTAGAAGC-3'); FasL (sense:
5'-GGATTGGGCCTGGGGATGTTTCA-3', antisense:
5'-TTGTGGCTCAGGGGCAGGTTGTTG-3'); perforin (sense:
5'-CAGGTCAACATAGGCATCCA-3', antisense: 5'-CGAGTTTACCCAGGCTGAGT-3'); granzyme B (sense: 5'-ACCTCTCCCAGTGTAAATCT-3', antisense:
5'-GCGGTGGCTTCCTGATACAA-3'); CD56 (sense:
5'-GAGATCAGCGTTGGAGAGTC-3', antisense: 5'-AAGAGTGACCTGCTCCTCTA-3'); -actin (sense: 5'-GTCCACCTTCCAGCAGATGT-3', antisense:
5'-CAATGCTATCACCTCCCCTG-3'). Reverse transcriptase (RT)-PCR products
for lambda-like 14.1, pre-T, and granzyme B have been cloned and
sequenced and corresponded to the sequences reported in public data
bank (not shown).
Cytokine production and virus activation Supernatants cryopreserved at the end of 6-day cultures with IL-3 were tested for IL-12 p70 and IL-8 contents by enzyme-linked immunosorbent assay (ELISA) (Beckman Coulter Immunotech; sensitivity, 5 pg/mL for IL-12 and 8 pg/mL for IL-8). To determine IFN- production, thawed purified tumor cells and FACS-sorted
CD56+ and CD56 cells from 3 patients (LAI,
GEN, and GUE) as well as fresh FACS-sorted blood, CD11c
pDCs, and CD11c+ myeloid-related DCs were stimulated with 1 hemagglutinating unit (HAU)/mL formaldehyde-inactivated influenza virus
strain Beijing/262/95 (kindly provided by Dr N. Kuehm, Aventis Pasteur,
Val de Reuil, France) in duplicate wells (106 cells/mL, in
24- or 96-well culture plate) for 24 hours. IFN- was
measured in supernatant by ELISA that specifically recognizes IFN-2
(Beckman Coulter Immunotech). The sensitivity of this assay is
0.6 U/mL.
Naive T-lymphocyte stimulation and polarization Proliferative response of naive T lymphocytes was evaluated in response to tumor cells activated or not. CD4+CD45RA+ lymphocytes were isolated from cord blood by negative immunomagnetic depletion (Stem Cell Technology, Meylan, France), resulting in more than 97% purity. Mixed lymphocyte cultures (MLCs) were conducted in quadruplicate in 200 µL 96-well flat-bottom plates (Falcon) by mixing 25 × 103 responding purified CD4+CD45RA+ cells and 5 × 103 to 25 × 103 irradiated (30 Gy) tumor cells (unactivated or after IL-3 treatment). Six-day cultures were performed in complete medium supplemented with 15% heat-inactivated human AB serum; 37 × 103 Bq [3H]thymidine was added to each well and harvested 18 hours later. Mo-DCs obtained from adherent peripheral blood mononuclear cells (normal volunteers) cultured for 6 days with GM-CSF (500 U/mL) and IL-13 (50 ng/mL)27 were used as control.To evaluate T-lymphocyte polarization, 5 × 103
irradiated IL-3-differentiated tumor cells were cocultured with
5 × 104 CD4+CD45RA+ cord blood T
cells during 6 days. The proportion of IL-4- and IFN-
Identification of a tumoral equivalent for pDCs Seven tumor samples were selected on the basis of their expression of CD4 and CD56 in the absence of other lineage markers (CD3, CD19, CD13, and CD33). Malignant cells expressed the panleukocyte antigen CD45 at a low level, similarly to blastic hematopoietic cells, but CD10, CD34, and CD117 that are related to progenitors were negative. Using a large panel of mAbs, we identified them as a homogeneous entity and postulated that they could be related to the pDC subset (Table 1). They were all positive for CD45RA and negative for CD45RO, CD1a, and CD11c (Table 1). CD1c, CD83, and CD116 were inconstantly slightly expressed (2 of 6, 1 of 7, and 3 of 7 cases, respectively). CD36 and CD68 monocytic antigens were always expressed (not shown), but CD14 and CD64 were absent. Regarding NK cell-related markers, besides CD56, granzyme B was detected both at the mRNA and protein levels (Figure 1 and Table 1). Intensity of granzyme B staining on leukemic pDCs (and normal pDCs, data not shown) was 10-fold lower than that observed on NK cells. Because perforin labeling was negative on leukemic pDCs (Table 1 and on normal pDCs, data not shown), we could not exclude that the slight mRNA signals for perforin and FasL on a leukemic sample (Figure 1) might be due to contaminating NK cells. Also, CD2 or CD7 could be expressed (1 of 7 and 4 of 7 cases, respectively), but CD16, CD57, CD94, and CD161 were negative (Table 1). These cells were also characterized by the presence of surface molecules associated with antigen-presenting cells (APCs): HLA-ABC and -DR in all cases, CD40 in 3 cases, and/or CD86 in 3 cases. They highly expressed immunoglobulin-like transcript (ILT)3 and IL-3R/CD123 (Table 1, Figure 1). Besides CD56, this phenotype is highly similar to
that of pDCs isolated from blood, tonsils, or thymus from nonmalignant patients. Like normal pDCs, tumor cells expressed strong levels of
pre-T-cell receptor (pre-T)16 and lambda-like
14.117 mRNA (Figure 1), both of which were negative
in NK cells and in mo-DCs. Of note, in all instances, no expression of
CD3 and CD19 mRNA was found, excluding T- and B-cell contaminations
(not shown).
Cell survival is sustained by IL-3 or GM-CSF Given the immunophenotypic similarities between these tumor cells and pDCs, we investigated the effect of IL-2, IL-3, IL-4, or GM-CSF on their in vitro survival (Figure 2). Day 0 viability of thawed cells was high (mean = 88%, not shown), but these cells died in the absence of cytokines (day 3 mean survival = 27%) even though slower than normal pDCs that undergo rapid spontaneous apoptosis in culture, resulting in 90% of cells displaying apoptotic figures at 16 hours.8 After a few hours in culture with IL-3 or GM-CSF, the tumor cells aggregated and formed large clusters, whereas their aspect remained unchanged with IL-2 or IL-4. IL-3 and GM-CSF improved slightly but significantly viability at day 3 from 27% to 52% and 48%, respectively (P < .05, Wilcoxon test). In contrast, IL-2 and IL-4 were not responsible either for such a significant improvement of cell survival or for an apoptotic signal. In 2 cases, day 3 [3H]thymidine incorporation was measured in the presence of these 4 cytokines, and no proliferation was detected (not shown).
Tumor cells differentiate into mature DCs with IL-3 and CD40L Six-day cultures of purified tumor cells were performed with IL-3 (5 cases) or GM-CSF (3 cases). As for normal pDCs, no increase in cell number was observed at the end of these cultures. In the presence of IL-3, the initial blastic morphology (Figure 3A) of the tumor cells was highly modified; the cells enlarged, acquiring an abundant cytoplasm containing many vacuoles (Figure 3B). A mature DC morphology was achieved with many fine dendrites when CD40L-transfected L cells were added on day 4 for 48 hours (Figure 3C). In the absence of cytokines, the cells that remained isolated rapidly died in culture (Figure 3D), whereas they formed large clusters in the presence of IL-3 (Figure 3E), in the periphery of which dendrites were observed upon CD40L-induced activation (Figure 3F). IL-3 induced in most cases an up-regulation of the expression of HLA-ABC and -DR molecules and of costimulatory molecules CD80 and CD86 (Table 2, Figure 4), which were further increased upon CD40 activation (Figure 4). CD1a, CD1c, and CD83 were also up-regulated in response to IL-3 alone (Table 2). In all cases but one (GEN), CD83 was negative on tumor cells (Table 1, Figure 3G). IL-3 led to a low surface expression of CD83 (Table 2), while high intracytoplasmic levels were detected in response to IL-3 (Figure 3H) and IL-3 plus CD40L (Figure 3I). In 3 of 5 cases, CD13 and/or CD33 appeared with IL-3, and CD11c became highly expressed in all cases. The morphology (not shown) and the immunophenotype of the cells differentiated with GM-CSF (3 cases, Table 2) were highly similar to the phenotype described with IL-3, and no expression of CD14 or CD64 could be detected. In 4 cases, we examined the profile of cytokines produced at the end of 6-day cultures with IL-3. As reported for CD40L-activated IL-3-matured pDCs,19 tumor cells produced IL-8 (mean = 1914 pg/mL), whereas IL-4 and IL-12 p70 were undetectable by ELISA (not shown).
DCs differentiated from leukemic cells stimulate allogeneic naive T cells and induce their Th2 polarization In the absence of activation, tumor cells from patients GEN and GUE did not induce the proliferation of naive T cells, whereas cells from patient CAT did. The ability of IL-3-differentiated leukemic DC to induce a primary allogeneic response was analyzed in MLC. In the 5 cases analyzed, a strong (8000 to 40 000 cpm) proliferation of cord blood-purified naive T cells was induced, similar to the proliferation obtained with mo-DCs (Figure 5, upper panels).
Given the reported ability of normal IL-3-matured pDCs to promote the
differentiation of Th2 cells,15,19 we next examined the
T-cell polarizing capacity of IL-3-differentiated leukemic DCs. After
a 6-day MLC, the cytokine production profile was analyzed (Figure 5,
lower panels). In 4 of 4 cases, the naive cord blood cells primed with
IL-3-differentiated leukemic DCs were mainly IL-4-producing cells
(9%-21%), whereas only a few IFN- Leukemic cells produce IFN- in response to virus.15,20 Thus, 4 patients (CAD, LAI, GEN, and GUE) were studied for the ability of either enriched tumor cells or FACS-sorted CD56+ and CD56
cells to produce IFN- in response to inactivated influenza virus. After a 24-hour incubation with the virus, but not upon CD40 signaling (not shown), significant levels of IFN- were measured in the supernatants of the culture from patients GEN and GUE that were purified from peripheral blood and lymph nodes, respectively, while no
IFN- production was detectable in those from bone marrow samples of
CAD and LAI. To exclude the possible contamination of enriched tumor
cells by residual normal pDCs, we FACS-sorted CD56+ and
CD56 cells (LAI, GEN, and GUE). Both the
CD56+ subset (devoid of normal pDCs) and CD56
subset from patients GEN and GUE produced IFN- in response to the
virus. Of note, (1) CD56 was not expressed homogeneously by leukemic cells (Table 1) and (2) we have shown that CD56
could be modulated in response to cytokines (Table
3). Thus, our interpretation is that most
of the enriched cells were indeed leukemic. Higher levels of IFN-
were produced by pDCs from healthy volunteers (24 990 IU/mL; Table 3)
compared with leukemic cells, which could be due to their malignant
status, their different organ origin, and/or an alteration of their
reactivity by cryopreservation.
In the present work, we analyzed unusual tumor cells in 7 patients. In light of recent knowledge from the literature, we
confirmed that they belong to the pDC lineage, because they
(1) produce IFN- The link between those tumor cells and pDCs has first been postulated
according to their unique phenotype: conventional T, B, and myeloid
lineage markers are absent, except CD7 or CD2 in some cases. T-cell and
B-cell receptors are in germline configuration, and no
myeloperoxidase enzymes are observed (not shown). Expression of
costimulatory molecules such as CD40 or CD86 in several cases is
evocative of an APC. Indeed, expression of CD4, HLA-DR, CD123, ILT3,
and CD45RA but absence of CD45R0 and CD11c is highly reminiscent of the
phenotype of pDCs.8,13,14 Moreover, despite the expression of some monocytic antigens like CD36 and CD68 (also reported on blood
CD11c Lymphoid-restricted progenitors exist in human bone marrow, thymus, and
fetal liver, the most immature one being able to differentiate into T,
B, and NK cells and DCs.10,11,33,34 However, those progenitors express CD34, which is not the case for leukemic pDCs. The
capacity of the malignant cells to differentiate into T, B, or NK cells
has to be tested, but at present we have shown that they are not
precursors of monocytes or macrophages. As for pDCs,13,14 culture in GM-CSF or IL-3 induced CD13, CD33, and CD11c myeloid antigens, but the interpretation with regard to the myeloid lineage is
not clear at present.8,35 Due to the expression of CD56 on
malignant cells, an NK origin has been evoked in many reports despite
the negativity of most NK differentiation markers and the absence of
azurophilic granulations. In fact, although classical for NK cells,
CD56 has often been described in malignant pathologies outside this
lineage, such as in myeloma36 and acute myeloid leukemia.37,38 Because of the positivity of granzyme B,
further functional assays are ongoing to assess the NK potential of
those leukemic cells. The down-regulation of CD56 expression observed after IL-3 activation in some cases suggests that the precursor involved in the leukemic transformation might be more immature than the
blood pDCs. We also consistently found genes associated with early T or
B precursors such as pre-T In conclusion, we demonstrate that a new hematologic entity has been
identified involving a malignant equivalent of plasmacytoid DCs. The
CD4+, CD56+, CD3
We thank Dr Xavier Ronot for help with contrast phase microscopy and Dr David Jarrossay for anti-ILT3 mAb. We thank Myriam Brachet, Patricia Cheron, Agnès Colomer, Ghislaine Del Vecchio, Richard Di Schiena, Michel Drouin, Jean-Paul Molens, Christine Vallet, and Mireille Favre for technical assistance. We thank Drs Dominique Masson and Agnès Moine for HLA typing of the cells. We thank Marina Cella, Marco Colonna, and Giorgio Trinchieri for advice and helpful discussions. We thank Christophe Caux for critically reading the manuscript and Isabelle Durand for FACS sorting.
Submitted August 14, 2000; accepted January 16, 2001.
M.C.J. and J.P. contributed equally to this work.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Laurence Chaperot or Marie-Christine Jacob, Dept of Research and Development, EFS Rhône-Alpes Grenoble, BP35, F38701 La Tronche Cedex, France; e-mail: laurence.chaperot{at}efs.sante.fr or marie-christine.jacob{at}efsrhonealpes.org.
1.
Kameoka J, Ichinohasama R, Tanaka M, et al.
A cutaneous agranular CD2 2. Adachi M, Maeda K, Takekawa M, et al. High expression of CD56 (N-CAM) in a patient with cutaneous CD4-positive lymphoma. Am J Hematol. 1994;47:278-282[Medline] [Order article via Infotrieve]. 3. Brody JP, Allen S, Schulman P, et al. Acute agranular CD4-positive natural killer cell leukemia: comprehensive clinicopathologic studies including virologic and in vitro culture with inducing agents. Cancer. 1995;75:2474-2483[CrossRef][Medline] [Order article via Infotrieve]. 4. DiGiuseppe JA, Louie DC, Williams JE, et al. Blastic natural killer cell leukemia/lymphoma: a clinicopathologic study. Am J Surg Pathol. 1997;21:1223-1230[CrossRef][Medline] [Order article via Infotrieve]. 5. Bagot M, Bouloc A, Charue D, Wechsler J, Bensussan A, Boumsell L. Do primary cutaneous non-T non-B CD4+CD56+ lymphomas belong to the myelo-monocytic lineage? J Invest Dermatol. 1998;111:1242-1244[CrossRef][Medline] [Order article via Infotrieve].
6.
Uchiyama N, Ito K, Kawai K, Sakamoto F, Takaki M, Ito M.
CD2 7. Petrella T, Dalac S, Maynadie M, et al. CD4+ CD56+ cutaneous neoplasms: a distinct hematological entity? Groupe Francais d'Etude des Lymphomes Cutanes (GFELC). Am J Surg Pathol. 1999;23:137-146[CrossRef][Medline] [Order article via Infotrieve].
8.
Grouard G, Rissoan MC, Filgueira L, Durand I, Banchereau J, Liu YJ.
The enigmatic plasmacytoid T cells develop into dendritic cells with interleukin (IL)-3 and CD40-ligand.
J Exp Med.
1997;185:1101-1111
9.
Galy A, Christopherson I, Ferlazzo G, Liu G, Spits H, Georgopoulos K.
Distinct signals control the hematopoiesis of lymphoid-related dendritic cells.
Blood.
2000;95:128-137
10.
Res P, Martinez Caceres E, Cristina Jaleco A, et al.
CD34+CD38dim cells in the human thymus can differentiate into T, natural killer, and dendritic cells but are distinct from pluripotent stem cells.
Blood.
1996;87:5196-5206
11.
Marquez C, Trigueros C, Fernandez E, Toribio ML.
The development of T and non-T cell lineages from CD34+ human thymic precursors can be traced by the differential expression of CD44.
J Exp Med.
1995;181:475-483 12. O'Doherty U, Peng M, Gezelter S, et al. Human blood contains two subsets of dendritic cells, one immunologically mature and the other immature. Immunology. 1994;82:487-493[Medline] [Order article via Infotrieve].
13.
Kohrgruber N, Halanek N, Groger M, et al.
Survival, maturation, and function of CD11c- and CD11c+ peripheral blood dendritic cells are differentially regulated by cytokines.
J Immunol.
1999;163:3250-3259 14. Robinson SP, Patterson S, English N, Davies D, Knight SC, Reid CD. Human peripheral blood contains two distinct lineages of dendritic cells. Eur J Immunol. 1999;29:2769-2778[CrossRef][Medline] [Order article via Infotrieve]. 15. Cella M, Jarrossay D, Facchetti F, et al. Plasmacytoid monocytes migrate to inflamed lymph nodes and produce large amounts of type I interferon. Nat Med. 1999;5:919-923[CrossRef][Medline] [Order article via Infotrieve].
16.
Res PC, Couwenberg F, Vyth Dreese FA, Spits H.
Expression of pTalpha mRNA in a committed dendritic cell precursor in the human thymus.
Blood.
1999;94:2647-2657
17. Nathalie Bendriss-Vermare, Clarisse Barthélémy, Isabelle
Durand, et al. Human thymus contains IFN
18.
Sorg RV, Kögler G, Wernet P.
Identification of cord blood dendritic cells as an immature CD11c- population.
Blood.
1999;93:2302-2307
19.
Rissoan MC, Soumelis V, Kadowaki N, et al.
Reciprocal control of T helper cell and dendritic cell differentiation.
Science.
1999;283:1183-1186
20.
Siegal FP, Kadowaki N, Shodell M, et al.
The nature of the principal type 1 interferon-producing cells in human blood.
Science.
1999;284:1835-1837 21. Cella M, Facchetti F, Lanzavecchia A, Colonna M. Plasmacytoid dendritic cells activated by influenza virus and CD40L drive a potent TH1 polarization. Nat Immunol. 2000;1:305-310[CrossRef][Medline] [Order article via Infotrieve].
22.
Kadowaki N, Antonenko S, Lau JY, Liu YJ.
Natural interferon alpha/beta-producing cells link innate and adaptive immunity.
J Exp Med.
2000;192:219-226
23.
Steinman R, Turley S, Mellman I, Inaba K.
The induction of tolerance by dendritic cells that have captured apoptotic cells.
J Exp Med.
2000;191:411-416
24.
Liu YJ, Blom B.
Introduction: TH2-inducing DC2 for immunotherapy.
Blood.
2000;95:2482-2483 25. Plumas J, Chaperot L, Jacob MC, et al. Malignant B lymphocytes from non-Hodgkin's lymphoma induce allogeneic proliferative and cytotoxic T cell responses in primary mixed lymphocyte cultures: an important role of co-stimulatory molecules CD80 (B7-1) and CD86 (B7-2) in stimulation by tumor cells. Eur J Immunol. 1995;25:3332-3341[Medline] [Order article via Infotrieve].
26.
Plumas J, Jacob MC, Chaperot L, Molens JP, Sotto JJ, Bensa JC.
Tumor B cells from non-Hodgkin's lymphoma are resistant to CD95 (Fas/Apo-1)-mediated apoptosis.
Blood.
1998;91:2875-2885 27. Chaperot L, Chokri M, Jacob MC, et al. Differentiation of antigen presenting cells (dendritic cells and macrophages) for therapeutic application in patients with lymphoma. Leukemia. 2000;14:1667-1677[CrossRef][Medline] [Order article via Infotrieve]. 28. Jacob MC, Favre M, Bensa JC. Membrane cell permeabilization with saponin and multiparametric analysis by flow cytometry. Cytometry. 1991;12:550-558[CrossRef][Medline] [Order article via Infotrieve].
29.
Strobl H, Scheinecker C, Riedl E, et al.
Identification of CD68+lin- peripheral blood cells with dendritic precursor characteristics.
J Immunol.
1998;161:740-748 30. Palucka K, Banchereau J. Linking innate and adaptive immunity. Nat Med. 1999;5:868-870[CrossRef][Medline] [Order article via Infotrieve].
31.
Hagmann M.
Elusive interferon alpha producers nailed down.
Science.
1999;284:1746
32.
Bottomly K.
T cells and dendritic cells get intimate.
Science.
1999;283:1124-1125 33. Galy A, Travis M, Cen D, Chen B. Human T, B, natural killer, and dendritic cells arise from a common bone marrow progenitor cell subset. Immunity. 1995;3:459-473[CrossRef][Medline] [Order article via Infotrieve].
34.
Plum J, De Smedt M, Verhasselt B, et al.
In vitro intrathymic differentiation kinetics of human fetal liver CD34+CD38- progenitors reveals a phenotypically defined dendritic/T-NK precursor split.
J Immunol.
1999;162:60-68
35.
Olweus J, BitMansour A, Warnke R, et al.
Dendritic cell ontogeny: a human dendritic cell lineage of myeloid origin.
Proc Natl Acad Sci U S A.
1997;94:12551-12556 36. Drach J, Gattringer C, Huber H. Expression of the neural cell adhesion molecule (CD56) by human myeloma cells. Clin Exp Immunol. 1991;83:418-422[Medline] [Order article via Infotrieve].
37.
Scott A, Head D, Kopecky K, et al.
HLA-DR, CD33+, CD56+, CD16- myeloid/natural killer cell acute leukemia: a previously unrecognized form of acute leukemia potentially misdiagnosed as French-American-British acute myeloid leukemia-M3.
Blood.
1994;84:244-255 38. Seymour JF, Pierce SA, Kantarjian HM, Keating MJ, Estey EH. Investigation of karyotypic, morphologic and clinical features in patients with acute myeloid leukemia blast cells expressing the neural cell adhesion molecule (CD56). Leukemia. 1994;8:823-826[Medline] [Order article via Infotrieve].
39.
McKenna HJ, Stocking KL, Miller RE, et al.
Mice lacking flt3 ligand have deficient hematopoiesis affecting hematopoietic progenitor cells, dendritic cells, and natural killer cells.
Blood.
2000;95:3489-3497 40. Lacombe F, Durrieu F, Briais A, et al. Flow cytometry CD45 gating for immunophenotyping of acute myeloid leukemia. Leukemia. 1997;11:1878-1886[CrossRef][Medline] [Order article via Infotrieve].
© 2001 by The American Society of Hematology.
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  A. J. Alencar, E. Bustinza, J. Barker, G. E. Byrne, and I. S. Lossos Hematodermic Tumor Presenting With Generalized Skin Involvement J. Clin. Oncol., June 20, 2009; 27(18): 3059 - 3061. [Full Text] [PDF]
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 M. Merad and M. G. Manz Dendritic cell homeostasis Blood, April 9, 2009; 113(15): 3418 - 3427. [Abstract] [Full Text] [PDF]
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 L. Fischer, N. Gokbuget, S. Schwartz, T. Burmeister, H. Rieder, M. Bruggemann, D. Hoelzer, and E. Thiel CD56 expression in T-cell acute lymphoblastic leukemia is associated with non-thymic phenotype and resistance to induction therapy but no inferior survival after risk-adapted therapy Haematologica, February 1, 2009; 94(2): 224 - 229. [Abstract] [Full Text] [PDF]
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T. Marafioti, J. C. Paterson, E. Ballabio, K. K. Reichard, S. Tedoldi, K. Hollowood, M. Dictor, M.-L. Hansmann, S. A. Pileri, M. J. Dyer, et al. Novel markers of normal and neoplastic human plasmacytoid dendritic cells Blood, April 1, 2008; 111(7): 3778 - 3792. [Abstract] [Full Text] [PDF]
|
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S. C.-S. Hu, K.-B. Tsai, G.-S. Chen, and P.-H. Chen Infantile CD4+/CD56+ hematodermic neoplasm Haematologica, September 1, 2007; 92(9): e91 - e93. [Full Text] [PDF]
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R. Dijkman, R. van Doorn, K. Szuhai, R. Willemze, M. H. Vermeer, and C. P. Tensen Gene-expression profiling and array-based CGH classify CD4+CD56+ hematodermic neoplasm and cutaneous myelomonocytic leukemia as distinct disease entities Blood, February 15, 2007; 109(4): 1720 - 1727. [Abstract] [Full Text] [PDF]
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D. Vremec, M. O'Keeffe, H. Hochrein, M. Fuchsberger, I. Caminschi, M. Lahoud, and K. Shortman Production of interferons by dendritic cells, plasmacytoid cells, natural killer cells, and interferon-producing killer dendritic cells Blood, February 1, 2007; 109(3): 1165 - 1173. [Abstract] [Full Text] [PDF]
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D. Allman, M. Dalod, C. Asselin-Paturel, T. Delale, S. H. Robbins, G. Trinchieri, C. A. Biron, P. Kastner, and S. Chan Ikaros is required for plasmacytoid dendritic cell differentiation Blood, December 15, 2006; 108(13): 4025 - 4034. [Abstract] [Full Text] [PDF]
|
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E. Chapiro, E. Delabesse, V. Asnafi, C. Millien, F. Davi, E. Nugent, K. Beldjord, T. Haferlach, D. Grimwade, and E. A. Macintyre Expression of T-lineage-affiliated transcripts and TCR rearrangements in acute promyelocytic leukemia: implications for the cellular target of t(15;17) Blood, November 15, 2006; 108(10): 3484 - 3493. [Abstract] [Full Text] [PDF]
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 L. Chaperot, A. Blum, O. Manches, G. Lui, J. Angel, J.-P. Molens, and J. Plumas Virus or TLR Agonists Induce TRAIL-Mediated Cytotoxic Activity of Plasmacytoid Dendritic Cells J. Immunol., January 1, 2006; 176(1): 248 - 255. [Abstract] [Full Text] [PDF]
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C.-W. Lin, T.-Y. Liu, S.-U. Chen, K.-T. Wang, L. J. Medeiros, and S.-M. Hsu CD94 1A transcripts characterize lymphoblastic lymphoma/leukemia of immature natural killer cell origin with distinct clinical features Blood, November 15, 2005; 106(10): 3567 - 3574. [Abstract] [Full Text] [PDF]
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 N. Bendriss-Vermare, S. Burg, H. Kanzler, L. Chaperot, T. Duhen, O. de Bouteiller, M. D'agostini, J.-M. Bridon, I. Durand, J. M. Sederstrom, et al. Virus overrides the propensity of human CD40L-activated plasmacytoid dendritic cells to produce Th2 mediators through synergistic induction of IFN-{gamma} and Th1 chemokine production J. Leukoc. Biol., October 1, 2005; 78(4): 954 - 966. [Abstract] [Full Text] [PDF]
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A. Fuchs, M. Cella, T. Kondo, and M. Colonna Paradoxic inhibition of human natural interferon-producing cells by the activating receptor NKp44 Blood, September 15, 2005; 106(6): 2076 - 2082. [Abstract] [Full Text] [PDF]
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B. Drenou, L. Amiot, N. Setterblad, S. Taque, V. Guilloux, D. Charron, R. Fauchet, and N. Mooney MHC class II signaling function is regulated during maturation of plasmacytoid dendritic cells J. Leukoc. Biol., April 1, 2005; 77(4): 560 - 567. [Abstract] [Full Text] [PDF]
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 B. Jahrsdorfer, L. Muhlenhoff, S. E. Blackwell, M. Wagner, H. Poeck, E. Hartmann, R. Jox, T. Giese, B. Emmerich, S. Endres, et al. B-Cell Lymphomas Differ in their Responsiveness to CpG Oligodeoxynucleotides Clin. Cancer Res., February 15, 2005; 11(4): 1490 - 1499. [Abstract] [Full Text] [PDF]
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F. Garnache-Ottou, L. Chaperot, S. Biichle, C. Ferrand, J.-P. Remy-Martin, E. Deconinck, P. D. de Tailly, B. Bulabois, J. Poulet, E. Kuhlein, et al. Expression of the myeloid-associated marker CD33 is not an exclusive factor for leukemic plasmacytoid dendritic cells Blood, February 1, 2005; 105(3): 1256 - 1264. [Abstract] [Full Text] [PDF]
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 K. McKenna, A.-S. Beignon, and N. Bhardwaj Plasmacytoid Dendritic Cells: Linking Innate and Adaptive Immunity J. Virol., January 1, 2005; 79(1): 17 - 27. [Full Text] [PDF]
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 M. W. Bekkenk, P. M. Jansen, C. J. L. M. Meijer, and R. Willemze CD56+ hematological neoplasms presenting in the skin: a retrospective analysis of 23 new cases and 130 cases from the literature Ann. Onc., July 1, 2004; 15(7): 1097 - 1108. [Abstract] [Full Text] [PDF]
|
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 CD56+ Lymphoma With Skin Involvement: Clinicopathologic Features and Classification Arch Dermatol, April 1, 2004; 140(4): 427 - 436.
|
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|
 |
|
 B. Vanbervliet, N. Bendriss-Vermare, C. Massacrier, B. Homey, O. de Bouteiller, F. Briere, G. Trinchieri, and C. Caux The Inducible CXCR3 Ligands Control Plasmacytoid Dendritic Cell Responsiveness to the Constitutive Chemokine Stromal Cell-derived Factor 1 (SDF-1)/CXCL12 J. Exp. Med., September 2, 2003; 198(5): 823 - 830. [Abstract] [Full Text] [PDF]
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M. Herling, M. A. Teitell, R. R. Shen, L. J. Medeiros, and D. Jones TCL1 expression in plasmacytoid dendritic cells (DC2s) and the related CD4+ CD56+ blastic tumors of skin Blood, June 15, 2003; 101(12): 5007 - 5009. [Abstract] [Full Text] [PDF]
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V. Asnafi, K. Beldjord, E. Boulanger, B. Comba, P. Le Tutour, M.-H. Estienne, F. Davi, J. Landman-Parker, P. Quartier, A. Buzyn, et al. Analysis of TCR, pTalpha , and RAG-1 in T-acute lymphoblastic leukemias improves understanding of early human T-lymphoid lineage commitment Blood, April 1, 2003; 101(7): 2693 - 2703. [Abstract] [Full Text] [PDF]
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M. R. Comeau, A.-R. Van der Vuurst de Vries, C. R. Maliszewski, and L. Galibert CD123bright Plasmacytoid Predendritic Cells: Progenitors Undergoing Cell Fate Conversion? J. Immunol., July 1, 2002; 169(1): 75 - 83. [Abstract] [Full Text] [PDF]
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D. Leroux, F. Mugneret, M. Callanan, I. Radford-Weiss, N. Dastugue, J. Feuillard, F. Le Mee, G. Plessis, P. Talmant, N. Gachard, et al. CD4+, CD56+ DC2 acute leukemia is characterized by recurrent clonal chromosomal changes affecting 6 major targets: a study of 21 cases by the Groupe Francais de Cytogenetique Hematologique Blood, May 13, 2002; 99(11): 4154 - 4159. [Abstract] [Full Text] [PDF]
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J. Feuillard, M.-C. Jacob, F. Valensi, M. Maynadie, R. Gressin, L. Chaperot, C. Arnoulet, F. Brignole-Baudouin, B. Drenou, E. Duchayne, et al. Clinical and biologic features of CD4+CD56+ malignancies Blood, March 1, 2002; 99(5): 1556 - 1563. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
-producing
cells was determined by flow cytometry after a further 6-hour
stimulation with 5 ng/mL phorbol myristate acetate (Sigma) and 0.5 µg/mL ionomycin (Sigma). During the last 5 hours, monensin (3 µM,
Sigma) was added. Cells were washed in cold phosphate-buffered saline,
fixed in paraformaldehyde (4%), and permeabilized with saponin
(0.1%).
