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HEMATOPOIESIS
From the Departments of Hematology and Rheumatology,
University Hospital of Crete School of Medicine, Heraklion, Greece; and
the Department of Hematology, St George's Hospital Medical School,
London, United Kingdom.
Based on previous reports for impaired hematopoiesis in rheumatoid
arhrtitis (RA), and in view of the current interest in exploring the
role of autologous stem cell transplantation (ASCT) as an alternative
treatment in patients with resistant disease, we have evaluated bone
marrow (BM) progenitor cell reserve and function and stromal cell
function in 26 patients with active RA. BM progenitor cells were
assessed using flow cytometry and clonogenic assays in short-term and
long-term BM cultures (LTBMCs). BM stroma function was assessed by
evaluating the capacity of preformed irradiated LTBMC stromal layers to
support the growth of normal CD34+ cells. We found that RA
patients exhibited low number and increased apoptosis of
CD34+ cells, defective clonogenic potential of BM
mononuclear and purified CD34+ cells, and low progenitor
cell recovery in LTBMCs, compared with healthy controls (n = 37).
Patient LTBMC stromal layers failed to support normal hematopoiesis and
produced abnormally high amounts of tumor necrosis factor alpha
(TNF Rheumatoid arthritis (RA) is a systemic autoimmune
disease characterized mainly by chronic destructive polyarthritis.
Although the main target tissue of the disease is the synovium,
evidence has been accumulated that bone marrow (BM) may be also
actively involved in the pathophysiology of RA by providing immune and inflammatory cells in the affected synovial membranes.1-3
It has been suggested that during the inflammatory stages of the disease, BM progenitor cells or even BM-derived immature mesenchymal cells enter the joint microenvironment through mechanisms similar to
those of mature inflammatory cells, and gradually replace normal synovial cells.4,5 Moreover, on the basis of findings from experimental models, it has been proposed that RA might even begin within the BM and might be regarded as a BM disorder.6,7 However, this hypothesis has not been adequately addressed in humans.
It is now well established that lymphocytes, macrophages, and
fibroblastlike cells found in rheumatoid synovium originate from or
have their counterpart in the BM. These cells are capable of producing
a variety of cytokines that may act not only as mediators of
inflammation but also as regulators of hematopoiesis. It is then
possible that BM, even if not primarily affected, may display impaired
function in RA as a consequence of abnormal interactions between immune
and hematopoietic cells or as a result of an aberrant, local or
systemic, cytokine production potentially affecting the survival,
proliferation, and optimal growth of hematopoietic cells. Indeed,
previous studies have shown an altered phenotype and abnormal growth
activity of BM myeloid progenitor cells in RA, at least at sites
adjacent to the affected joints.2,8-10 It has also been reported that elevated levels of locally produced proinflammatory cytokines, such as tumor necrosis factor alpha (TNF However, stem cells per se, in terms of their number and
functional characteristics, and BM microenvironment, in terms of its
capacity to support hematopoietic progenitor cell growth, have not been
extensively studied in RA. In view of the current interest in exploring
the use of high-dose immunosuppression followed by autologous
hematopoietic stem cell transplantation (ASCT) in patients with severe,
resistant autoimmune diseases including RA,13-16 it is
important to know whether BM stem cell compartment and/or BM
microenvironment are already affected by the inflammatory process in
these patients. The aim of the present study was to evaluate
hematopoietic progenitor cell reserve and function and BM stromal cell
hematopoiesis supporting capacity in patients with severe, active RA.
Patients
BM samples
Immunophenotyping and 7-amino-actinomycin D staining An indirect immunofluorescence technique was used to quantitate BM CD34+ cells. In brief, 1 × 106 BMMCs were stained with phycoerythrin (PE)-conjugated mouse antihuman CD34 Mab (QBEND-10; Immunotech, Marseille, France) and fluorescein isothiocyanate (FITC)-conjugated mouse antihuman CD38 Mab (T16; Immunotech) for 30 minutes on ice. PE- and FITC-conjugated mouse IgG isotype-matched controls were used as negative controls. Cells were washed twice in phosphate buffered saline (PBS) containing 1% fetal calf serum (FCS; Gibco BRL, Life Technologies) and 0.05% azide, and fixed in 500 µL 2% paraformaldeyde solution (Sigma).Aliquots of 1 × 106 BMMCs stained with PE-conjugated anti-CD34 Mab and FITC-conjugated mouse antihuman Fas (CD95) Mab (LOB 3/17; Serotec, United Kingdom) as above, were further stained prior to fixation with 7-amino-actinomycin D (7AAD; Calbiochem-Novabiochem, La Jolla, CA) as previously described.21 In brief, 100 µL 7AAD solution (200 µg/mL) was added to the cells, suspended in 1 mL PBS, and incubated for 20 minutes on ice protected from the light. Following centrifugation, the supernatant was removed and cells were fixed as above. Unstained fixed cells were used as negative controls. Cell samples were analyzed on an Epics Elite model flow cytometer
(Coulter, Miami, FL) within 30 minutes of fixation. Data were acquired
and processed on 500 000 events. After creating a scattergram
combining forward and right-angle light scatter for the whole
population, a region was drawn around BMMCs (low forward and low right
scatter properties) for CD34+ and CD38+ cell
estimation. When cells were stained for CD34 antigen, Fas antigen, and
7AAD, a scattergram was created by combining right-angle light scatter
with CD34 fluorescence in the gate of BMMCs and a second scattergram
was generated by combining CD34 and Fas fluorescence in the gate of
CD34+ cells. Finally, a scattergram was generated by
combining forward light scatter with 7AAD fluorescence to quantitate
7AAD-negative (live), -dim (apoptotic), and -bright (dead) cells in the
gate of CD34+, CD34+/Fas+, and
CD34+/Fas
Purification of CD34+ cells CD34+ cells were isolated from BMMCs by indirect magnetic labeling (magnetic activated cell sorting; MACS isolation kit, Mitenyi Biotec GmbH, Bergisch Gladbach, Germany) according to the manufacturer's protocol. In each experiment, purity of CD34+ cells was more than 96% as estimated by flow cytometry.Clonogenic progenitor cell assays BMMCs or CD34+ BM cells were cultured in 1 mL IMDM supplemented with 30% FCS, 1% bovine serum albumin (BSA; Gibco BRL Life Technologies), 10 4 M mercaptoethanol (Sigma),
0.075% sodium bicarbonate (Gibco BRL Life Technologies), 2 mM
L-glutamine (Sigma), 0.9% methylcellulose (StemCell
Technologies, Vancouver, BC, Canada), in the presence of 5 ng
granulocyte macrophage-colony-stimulating factor (GM-CSF; R&D Systems,
Minneapolis, MN), 50 ng interleukin-3 (IL-3; R&D Systems), and 2 IU
erythropoietin (EPO; Janssen-Ciliag, Bucks, United Kingdom), at
a concentration of 105 BMMCs or 3 × 103
CD34+ cells/mL of culture medium. Cultures were set up in
duplicate in 35-mm petri dishes and incubated at a 37°C, 5%
CO2, fully humidified atmosphere. On day 14, colonies were
scored and classified as granulocyte colony-forming units (CFU-G),
macrophage colony-forming units (CFU-M), granulocyte-macrophage
colony-forming units (CFU-GM), erythroid burst-forming units (BFU-E)
and colonies containing both granulocyte-macrophage and erythroid
elements (CFU-GEM) according to established criteria.22
Results were expressed as total numbers of CFU-GM (CFU-G plus CFU-M
plus CFU-GM) and total numbers of BFU-E (BFU-E plus CFU-GEM).
Long-term bone marrow cultures Long-term bone marrow cultures (LTBMCs) from 107 BMMCs were grown according to the standard technique23,24 in 10 mL IMDM supplemented with 10% FCS, 10% horse serum (Gibco BRL Life Technologies), 100 IU/mL PS, 2 mmol L-glutamine and 106 mol hydrocortisone sodium succinate (Sigma), and
incubated at a 33°C, 5% CO2, fully humidified
atmosphere. At weekly intervals, cultures were examined for stromal
layer formation using an inverted microscope and fed by removing half
of the medium and replacing it with equal volume of fresh IMDM
supplemented as above. Nonadherent cells were counted and assayed for
CFU-GM and BFU-E committed progenitor cells as described above. Colony
results were expressed as total numbers of colony-forming cells (CFCs)
(CFU-GM plus BFU-E). When mentioned, a mouse antihuman TNF
monoclonal neutralizing antibody (R&D Systems) was added weekly to the
cultures at a concentration of 1.8 µg/mL. According to the
manufacturer, the neutralization dose50 for this
antibody is 0.02 µg/mL to 0.04 µg/mL in the presence of 0.25 ng/mL
TNF. On confluence (week 3-4), cell-free supernatants were harvested
and stored at  70°C for TNF quantification by means of a
commercially available enzyme-linked immunosorbent assay (ELISA) kit
(Biosource International, Camarillo, CA). According to the
manufacturer, the sensitivity of this assay is less than 0.09 pg/mL.
Assessment of BM stromal cell function A 2-stage culture procedure was used to test the capacity of BM stromal layers to support normal hematopoiesis. Confluent stromal layers from patients and healthy controls, grown in standard LTBMCs, were irradiated (10 Gy) and recharged with 5 × 104 normal allogeneic CD34+ BM cells as previously described.25 In each experiment, flasks were recharged in triplicate and the CD34+ cells from the same healthy control were used to test patient and healthy control cultures. Cultures were fed weekly by demidepopulation and supernatants were monitored by determining the total number of nonadherent cells and CFC frequency.Statistical analysis Data were analyzed in the GraphPad Prism statistical PC program (GraphPad Software, San Diego, CA) by means of the nonparametric Mann-Whitney test, the Student t test for paired samples, and the Pearson coefficient of correlation test. Standard 2-way analysis of variance test was used to define differences in the number of nonadherent cells and the number of CFCs generated in LTBMCs in the groups tested. The F value represents the ratio of variance between the groups and the total variance of values in the study. Statistically significant differences for a certain F value are given in specific tables on the basis of the respective degrees of freedom.26
Flow-cytometric data The percentage of BM CD34+ cells and their subpopulations in patients and healthy controls are presented in Table 2. Patients with RA displayed significantly lower proportions of CD34+ cells within the BMMCs, compared with the controls (P = .0016). This decrease probably reflected the reduction of the committed CD34+/CD38+ cells (P = .0017), because no statistically significant difference was found between patients and healthy controls in the number of the more primitive CD34+/CD38 cells
(P = .1890).
To explore whether the decrease of CD34+ cells in patients with RA was due to increased apoptotic cell death, we evaluated the proportion of apoptotic cells within the CD34+ cell fraction using 7AAD staining (Table 2). We found that patient CD34+ cells contained significantly higher numbers of apoptotic (7AADdim) cells compared with the controls (P = .0136). In contrast, no statistically significant difference was found between patients (6.96% ± 5.86%, n = 23) and control subjects (6.69% ± 5.61%, n = 24) in the percentage of apoptotic cells detected in the non-CD34+ BMMC fraction (P = .6630). Because Fas antigen expression has been associated with apoptosis of BM
hematopoietic progenitor cells,27 we evaluated the expression of this molecule on patients' CD34+ cells
(Table 2). We found that the proportion of Fas+ cells in
the gate of CD34+ cells was significantly higher in the
group of patients than in the group of controls
(P = .0210). A highly significant positive correlation was
noted between the proportion of Fas+ cells and the
percentage of apoptotic cells within the CD34+ cell
population in the entire group of subjects studied (r = 0.611,
P < .0001). Moreover, the increased apoptosis found in patients' CD34+ cell compartments was due mainly to the
increased proportion of apoptotic cells within the
CD34+/Fas+ cell fraction
(P = .0310), because no statistically significant difference could be demonstrated between patients and healthy controls
in the proportion of apoptotic cells within the
CD34+/Fas Because the majority of the patients had been previously treated with cytotoxic and/or immune suppressant agents, a subset analysis in the number of CD34+, Fas+, and apoptotic cells in the groups of previously treated (n = 20) and untreated (n = 6) patients was performed, to exclude the possibility of drug-induced damage. Statistical analysis was similar in treated and untreated patients (Table 2), suggesting that drug-induced damage of patient progenitor cells is unlikely. Colony-forming cells The frequency of clonogenic progenitor cells in the BMMC fraction of patients with RA (n = 25) and healthy controls (n = 25) is depicted in Figure 2. The mean number of CFU-GM and BFU-E obtained by 107 BMMCs was significantly lower in the patients (4648 ± 2197 and 1923 ± 1286, respectively) compared with the controls (7680 ± 3603 and 4196 ± 1859, respectively; P < .002 and P < .0001 respectively). Further analysis showed that compared with the healthy controls, the numbers of CFU-GM and BFU-E were significantly lower in both the previously treated (n = 19) (5284 ± 2079 per 107 BMMCs; P = .0007 and 2169 ± 1368 per 107 BMMCs; P = .0005, respectively) and untreated (n = 6) (2633 ± 1097 per 107 BMMCs; P = .0209 and 1150 ± 769 per 107 BMMCs; P = .0002, respectively) groups of patients, suggesting further that drug-induced damage concerning the number and/or the clonogenic potential of patient progenitor cells is unlikely.
Individual CFC values correlated with the proportion of CD34+ cells in the group of controls (r = .508, P < .01) but not in the group of patients with RA (r = .196, P = .359), suggesting a possible defect in the clonogenic potential of patient progenitor cells. To further investigate this hypothesis, we evaluated the colony-forming potential of immunomagnetically purified CD34+ cells from patients with RA (n = 12) (Figure 2). Indeed, CFU-GM and BFU-E colony formation by 5 × 104 CD34+ cells was significantly lower in the patient group (324 ± 241 and mean 147 ± 88, respectively) than in the group of controls (n = 21) (544 ± 308 and 310 ± 107, respectively; P = .036 and P = .0005, respectively). Standard LTBMCs Typical stromal layers consisting of fibroblastlike cells, macrophages, and cobblestone areas were formed over the first 3 weeks in both patient (n = 25) and normal (n = 25) LTBMCs. However, stromal layers from patients with RA reached confluence earlier than those from the healthy controls (F = 12.487 > F    
Recharged LTBMCs To further evaluate the hematopoiesis-supporting capacity of patient stromal cells independently of stem cell function, confluent stromal layers from 8 patients with RA and 5 healthy controls were recharged with normal CD34+ cells. Patient LTBMC stromal layers failed to support normal hematopoiesis as indicated by the average total number of nonadherent cells (F = 10.908 > F TNF in the supernatants of confluent LTBMCs
(weeks 3-4) were determined in all patients and 11 healthy
controls (Figure 4). Patients with RA
displayed significantly higher cytokine concentrations (17.87 ± 12.01 pg/mL, median 15.98 pg/mL, range 3.21 to 55.00 pg/mL) compared
with the healthy subjects (6.28 pg/mL ± 2.41 pg/mL, median 6.93 pg/mL, range 1.37 to 9.40 pg/mL; P = .0005). Individual
TNF values inversely correlated with the proportion of
CD34+ cells (r = 0.513; P = .0016) and the
total number of CFCs (r = 0.349; P = .040), and
positively with the proportion of CD34+/Fas+
cells (r = 0.703; P < .001) and the proportion of
CD34+/7AADdim cells (r = .732;
P < .001) in the entire group of subjects studied (Figure
5). These data suggest that the increased
TNF production by patient BM stromal cells probably accounts for the
quantitative and functional abnormalities found in patients'
progenitor cell compartments. However, experiments in which
1 × 106 normal BMMCs were incubated for 48 hours in 1 mL
IMDM 20% FCS in the absence or presence of human rhTNF (R&D
Systems) at a concentration of 6.28 pg/mL and 17.87 pg/mL, representing
the mean TNF value found in LTBMC supernatants of healthy subjects and patients with RA respectively, did not show any difference in the
expression of Fas within the CD34+ cell population (data
not shown). These findings, however, do not reverse our suggestion
regarding the role of TNF on hematopoiesis in RA since the in vivo
conditions are different and yet unknown. In fact, we do not know the
real concentration of TNF within the BM microenvironment in soluble
and membrane-bound form, or the interactions of the cytokine with other
factors locally produced.11 It is probable that higher
concentrations of TNF are required for the in vitro demonstration of
the cytokine effect on the expression of Fas on CD34+
progenitor cells.28
Effect of anti-TNF treatment and results were compared to pretreatment values (Table 3). A significant increase was found in
the percentage of CD34+ cells following treatment
(P = .003), probably reflecting the increase of the
committed CD34+/CD38+ cells
(P = .004), because no significant changes were noted in the proportion of early CD34+/CD38 cells
(P = .696). The proportion of apoptotic cells within the CD34+ cell compartment was found to be significantly
reduced after anti-TNF treatment (P = .016), and this
reduction was associated with a significant decrease in the proportion
of CD34+ cells expressing Fas antigen
(P = .029). Further analysis showed a significant decrease
in the percentage of apoptotic cells detected in the
CD34+/Fas+ cell fraction
(P = .009) but not in the
CD34+/Fas cell fraction
(P = .972) following treatment.
Furthermore, the number of CFCs obtained by 107 BMMCs in
the clonogenic assays after anti-TNF
Finally, to investigate the possible changes in the capacity of BM
stroma to support hematopoiesis after anti-TNF Effect of anti-TNF production by BM stromal
cells accounts for the impaired hematopoiesis in RA, we also tested the
effect of the exogenous addition of an anti-TNF neutralizing antibody on the colony formation in LTBMCs. We used the antibody at a
concentration 100-fold higher than the highest TNF value found in
patient LTBMC supernatants (55 pg/mL). In patients with RA studied
prior to in vivo treatment with anti-TNF (n = 7), the exogenous
weekly addition of anti-TNF in LTBMCs significantly increased the
number of nonadherent cells and the CFCs compared to their untreated
LTBMCs (F = 4.845 > F  treatment (n = 3), no
statistically significant difference was found in the number of
nonadherent cells or the CFC recovery in LTBMCs treated with
anti-TNF compared with their untreated cultures (F = .574 < F  in hematopoiesis in
patients with RA.
Regulation of hematopoiesis and maintenance of homeostasis in BM
requires a well-balanced interaction between the hematopoietic cells
and the immune system. It has been shown that immune dysregulation in
autoimmune and chronic inflammatory disorders may modulate the function
of BM hematopoietic progenitor cells and/or their microenvironment,
either by inflammatory cytokine production or by intricate cell-to-cell
interactions.29 RA is a chronic systemic inflammatory
disease of autoimmune origin, which has previously been associated with
a variety of functional and immunophenotypic abnormalities in BM
attributable mainly to augmented local production of inflammatory
cytokines30,31 or increased T-cell
activation,32,33 or even to an intrinsic or
drug-induced34 hematopoietic or stroma abnormality.6,7 This study describes a significant
quantitative and functional hematopoietic progenitor cell defect in
patients with active RA indicated by the low number of
CD34+ cells, the impaired clonogenic potential of BM
progenitor cells, and the decreased progenitor cell recovery in LTBMCs.
These abnormalities may represent either an intrinsic progenitor cell
defect or secondary progenitor cell damage in response to an underlying
inflammatory process within the BM microenvironment. Indeed, data from
this study showed that patient BM stromal cells produced abnormally high amounts of TNF Using flow cytometry we found that the proportion of apoptotic cells
within patients' CD34+ cell compartments was significantly
increased and was associated with a significant up-regulation of Fas
antigen. Although Fas antigen expression does not always lead to
apoptotic cell death,35 it seems likely that the Fas
pathway is actively involved in the apoptotic cell depletion of patient
CD34+ cells since a highly significant correlation was
found between the proportions of Fas+ and apoptotic
CD34+ cells and a negative correlation was found between
the percentage of CD34+ cells and the proportion of
Fas+ progenitor cells in our patients. In keeping with this
aspect is the observation that the percentage of apoptotic cells was significantly higher among the Fas+ rather than among the
Fas Colony formation by BMMCs was defective in our patients in both
short-term and long-term cultures. Although one could postulate that
this defect might simply reflect the lower proportion of CD34+ cells in patients' BMMC fractions, this hypothesis
seems unlikely since the colony-producing cell number did not correlate
with the proportion of CD34+ cells in the group of
patients, suggesting a possible intrinsic defect in the clonogenic
potential of patient progenitor cells. The reduced colony formation by
highly purified patient CD34+ cells is consistent with this
assumption. The underlying mechanism(s) responsible for this
abnormality is unclear; however, since CFC numbers correlated inversely
with TNF The long-term exposure to antirheumatic and cytotoxic agents such a methotrexate might be a contributing factor affecting BM progenitor cell reserve and function in patients with RA. However, it has been shown that the myelosuppression occasionally seen during treatment with methotrexate affects exclusively late stages of hematopoietic development but not the early myeloid and progenitor cells.39,40 Immunosuppressive agents including glucocorticoids have been reported to induce apoptosis in mitogen-induced peripheral blood mononuclear cells41; however, there is no available evidence that they may affect the BM stem cell compartment. In contrast, there are reports suggesting a rather positive effect of glucocorticoids in the proliferation and differentiation of BM progenitor cells.42-44 On the other hand, 6 of our patients, suggesting a proportion of 23.1%, were studied prior to exposure to cytotoxic agents or immune suppressants, indicating that drug-induced damage is not the major factor affecting hematopoiesis in patients with RA. In keeping with this assumption is the low number of CD34+ cells, the increased apoptosis, and expression of Fas antigen in the CD34+ compartment and the low progenitor cell recovery in short-term and long-term BM cultures in the subgroup of patients who had never been previously treated with immune suppressants compared with healthy subjects. This study also reports impaired BM stroma function in RA, indicated by
the reduced capacity of patient stromal layers to support the growth of
autologous or allogeneic normal hematopoietic progenitors in an LTBMC
system. In addition to the increase of CD34+ cell number,
the anti-TNF Patient LTBMC stromal layers reached confluence earlier than did those
of the healthy controls. This finding is in agreement with previous
reports, suggesting increased proportion and proliferative capacity of
cells of monocyte-macrophage lineage in the BMMC fraction of patients
with RA.10,45 These cells primarily participate in the
adherent layer formation in LTBMC assays and their synovial counterparts are considered to be the central inflammatory cells producing TNF In conclusion, data from this study suggest that patients with active
RA exhibit low frequency and accelerated apoptosis of BM
CD34+ cells, defective clonogenic potential of marrow
progenitor cells, and impaired hematopoiesis-supporting capacity of BM
stroma. We suggest that these abnormalities may be due, at least in
part, to the increased TNF
We would like to thank Dr Maria Psyllaki and Dr Charis Ponticoglou for their technical assistance and the rheumatology, internal medicine, and orthopedics clinical staff of the University Hospital of Heraklion for providing BM samples from patients with RA and from healthy controls.
Submitted June 4, 2001; accepted October 26, 2001.
Supported by University Hospital of Heraklion, Greece.
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: Helen A. Papadaki, Department of Hematology, University Hospital of Heraklion, PO Box 1352, Heraklion, Crete, Greece; e-mail: epapadak{at}med.uoc.gr.
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© 2002 by The American Society of Hematology.
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  S. D. Yogesha, S. M. Khapli, R. K. Srivastava, L. S. Mangashetti, S. T. Pote, G. C. Mishra, and M. R. Wani IL-3 Inhibits TNF-{alpha}-Induced Bone Resorption and Prevents Inflammatory Arthritis J. Immunol., January 1, 2009; 182(1): 361 - 370. [Abstract] [Full Text] [PDF]
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J Larghero, D Farge, A Braccini, S Lecourt, A Scherberich, E Fois, F Verrecchia, T Daikeler, E Gluckman, A Tyndall, et al. Phenotypical and functional characteristics of in vitro expanded bone marrow mesenchymal stem cells from patients with systemic sclerosis Ann Rheum Dis, April 1, 2008; 67(4): 443 - 449. [Abstract] [Full Text] [PDF]
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P. E Westerweel, R. K M A C Luijten, I. E Hoefer, H. A Koomans, R. H W M Derksen, and M. C Verhaar Haematopoietic and endothelial progenitor cells are deficient in quiescent systemic lupus erythematosus Ann Rheum Dis, July 1, 2007; 66(7): 865 - 870. [Abstract] [Full Text] [PDF]
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L. P. Mueller, J. Luetzkendorf, T. Mueller, K. Reichelt, H. Simon, and H.-J. Schmoll Presence of Mesenchymal Stem Cells in Human Bone Marrow After Exposure to Chemotherapy: Evidence of Resistance to Apoptosis Induction Stem Cells, December 1, 2006; 24(12): 2753 - 2765. [Abstract] [Full Text] [PDF]
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L. M. Sedger, S. Hou, S. R. Osvath, M. B. Glaccum, J. J. Peschon, N. van Rooijen, and L. Hyland Bone Marrow B Cell Apoptosis During In Vivo Influenza Virus Infection Requires TNF-{alpha} and Lymphotoxin-{alpha} J. Immunol., December 1, 2002; 169(11): 6193 - 6201. [Abstract] [Full Text] [PDF]
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H. A. Papadaki, H. D. Kritikos, V. Valatas, D. T. Boumpas, and G. D. Eliopoulos Anemia of chronic disease in rheumatoid arthritis is associated with increased apoptosis of bone marrow erythroid cells: improvement following anti-tumor necrosis factor-alpha antibody therapy Blood, June 28, 2002; 100(2): 474 - 482. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
isotype,
with specificity for recombinant and natural human
TNF
); healthy
controls, (
).
and has
been associated with bone marrow failure syndromes due to accelerated
stem cell apoptosis.