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Prepublished online as a Blood First Edition Paper on June 14, 2002; DOI 10.1182/blood-2002-02-0558.
NEOPLASIA
From the Departments of Immunology, Pediatric and
Adolescent Medicine, and Internal Medicine, Mayo Graduate and Medical
Schools, Mayo Clinic, Rochester, MN.
B-cell chronic lymphocytic leukemia (B-CLL) is defined by the
accumulation of CD5+ B cells in the periphery and bone
marrow. This disease is not characterized by highly proliferative cells
but rather by the presence of leukemic cells with significant
resistance to apoptosis and, therefore, prolonged survival.
B-lymphocyte stimulator (BLyS) is a newly identified tumor necrosis
factor (TNF) family member shown to be critical for maintenance of
normal B-cell development and homeostasis and it shares significant
homology with another TNF superfamily member, APRIL. The striking
effects of BLyS on normal B-cell maintenance and survival raises the
possibility that it may be involved in pathogenesis and maintenance of
hematologic malignancies, including B-CLL. In this study, we
investigated the status of APRIL and BLyS expression, as well as their
receptors, in this disease. All B-CLL patient cells studied expressed
one or more of 3 known receptors for BLyS; however, the pattern of expression was variable. In addition, we demonstrate for the first time
that B-CLL cells from a subset of patients aberrantly express BLyS and
APRIL mRNA, whereas these molecules were not detectable in normal B
cells. Furthermore, we provide in vitro evidence that BLyS protects
B-CLL cells from apoptosis and enhances cell survival. Because these
molecules are key regulators of B-cell homeostasis and tumor
progression, leukemic cell autocrine expression of BLyS and APRIL may
be playing an important role in the pathogenesis of this disease.
(Blood. 2002;100:2973-2979) B-cell chronic lymphocytic leukemia (B-CLL)
is the most common leukemia in the Western world and is defined by the
accumulation of CD5+ B cells with prolonged survival in the
periphery and bone marrow.1,2 One of the current
challenges to a better understanding of this disease is the extensive
clinical and biologic heterogeneity found among patients with B-CLL.
B-CLL cells are defined by a unique set of cell surface molecules:
CD19+, CD5+, CD23+, and low levels
of surface immunoglobulin (Ig). However, additional biologic features
permit further subcharacterization of B-CLL, including Ig heavy-chain
variable region somatic mutation status and CD38 expression
levels.1,2 Although numerous advances have been made in
our understanding of B-CLL, the molecular pathogenesis of this disease
remains to be elucidated. However, it has been demonstrated that
certain molecular alterations, including overexpression of
antiapoptotic proteins and recurring chromosomal abnormalities, can be
found regularly in B-CLL.1,2
In this regard, members of the tumor necrosis factor (TNF) superfamily
have been shown to be key mediators in the formation and regulation of
normal B-cell responses.3 B-lymphocyte stimulator, BLyS,4 also called B cell-activating factor
(BAFF),5 TNF homologue that activates apoptosis, nuclear
factor Three receptors, B-cell maturation antigen (BCMA),20
transmembrane activator and CAML interactor (TACI),21 and
BAFF-R22 have been identified as receptors for BLyS. BCMA
and BAFF-R are exclusively expressed on B lymphocytes, whereas TACI can
be found on B cells and activated T cells. TACI and BCMA can also bind to APRIL, whereas BAFF-R is specific for BLyS. A key role for BAFF-R in
BLyS binding has been suggested by studies demonstrating that A/WySnJ
mice, which carry a mutation in the BAFF-R, have a loss of follicular
and marginal zone B cells in secondary lymphoid organs, a phenotype
similar to BLyS-deficient mice.10,22-24 Thus, BAFF-R
appears to be the primary receptor for BLyS responsible for B-cell
development and survival. However, TACI-deficient mice were found to
have an accumulation of splenic B cells and TACI Although there appears to be an essential role for BLyS in B-cell
development and survival, the precise mechanism of BLyS action is less
clearly understood. Treatment of A20 lymphoma cells with BLyS results
in NF- The striking effects of BLyS on normal B-cell maintenance and survival
raises the possibility that the BLyS-TACI/BCMA/BAFF-R receptor system
may be involved in pathogenesis and maintenance of B-cell malignancies.
Accordingly, we began an investigation of the status of APRIL and BLyS
expression in B-CLL. Of great interest, we show for the first time that
B-CLL cells from a subset of patients aberrantly express BLyS and
APRIL. In addition, exogenous BLyS promoted B-CLL cell survival and
soluble decoy receptors enhanced B-CLL apoptosis.
Cells and reagents
PCR analysis
Real-time PCR was performed on RNA samples isolated from CD19+ B-CLL cells, HL60 myelomonocytic cells, or HT2260 and Raji B lymphoblastic cells. HeLa RNA was purchased from BD Pharmingen (Franklin Lakes, NJ). Quantitative PCR was carried out with the use of double fluorescently labeled probes (synthesized by Applied Biosystems, Foster City, CA) specific for cyclophilin-B or BLyS. The probes were labeled at the 5' end with the fluorescent reporter dye VIC and at the 3' end with tetramethylrhodamine (TAMARA) as the quencher. cDNA synthesis and quantitative PCR was performed in a single 50-µL reaction containing 25 µL OneStep RT-PCR Master Mix containing AmpliTaq Gold DNA Polymerase (Applied Biosystems), 1.25 µL 40 × MultiScribe reverse transcriptase (Applied Biosystems), 125 mM sequence specific hybridization probes for BLyS (VIC-CCA CCA GCT CCA GGA GAA GGC AAC TC-TAMRA) or cyclophilin-B mRNA (VIC-AGC ATC TAC GGT GAG CGC TTC CCC-TAMRA), 300 nM forward and reverse primers for BLyS (5' CGC GGG ACT GAA AAT CTT TG and 3' CAC GCT TAT TTC TGC TGT TCT GA) or cyclophilin-B (5' GGA GAT GGC ACA GGA GGA AA and 3' CGT AGT GCT TCA GTT TGA AGT TCT CA), and 1 to 5 µg RNA. cDNA synthesis and RT-PCR were performed using the ABI Prism 7000 Sequence Detection System (Applied Biosystems) with the following thermal cycler protocol. Step 1: 48°C for 30 minutes; step 2: 95°C for 10 minutes; step 3: 40 cycles of denaturation at 95°C for 15 seconds and annealing at 60°C for 1 minute. Quantitative PCR analysis was completed using ABI PRISM 7000 SDS
Software. C Caspase-3 activity assay B-CLL cells were isolated, washed, and cultured (10 × 106 cells/mL) in phenol red-free RPMI supplemented with 0.5% bovine serum albumin (BSA) and 0.1 µg/mL Flag-BLyS, 10 µM chlorambucil, 10 µg/mL human IgG, Fc fragment, or 1 µg/mL BCMA-Fc at 37°C. After 18 hours of incubation, cells were washed and plated into a 96-well plate (5 × 106 cells/well in 25 µL phenol red-free RPMI in triplicate). Select wells were incubated with the caspase-3-specific inhibitor Ac-DEVD-CHO (50 nM final; Biomol, Plymouth Meeting, PA) for 30 minutes at 37°C prior to addition of 50 µL of the caspase-3-specific fluorogenic substrate Ac-DEVD-AMC (33 µM final; Biomol) in substrate buffer (100 mM HEPES [N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid], 10% sucrose, 0.1% CHAPS (3[3-chloroaminopropyl diethyl-ammonio[-1-propane sulfonate], 5 mM DTT [dithiothreitol], and 0.0001% NP40). Caspase-3 activity was measured by cleavage of the caspase-3-specific substrate and was assessed by fluorescence using a CytoFluor Multi-well Plate Reader (Applied Biosystems).Western blot analysis B-CLL cells, stimulated as described above, were directly lysed in 150 µL Laemmli sample buffer. Cell lysates (10 × 106 cells/lane) were boiled, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and proteins were transferred to an Immobilon P membrane (Millipore, Bedford, MA). Membranes were incubated with 1 µg/mL anti-PARP followed by horseradish peroxidase (HRP)-linked goat antimouse secondary antibody. Immunoreactive proteins were detected using enhanced chemiluminescence (Pierce, Rockford, IL).
Expression of TACI, BCMA, and BAFF-R in CLL B cells Initially, we examined TACI expression by B-CLL cells and normal B cells. Using fluorescence-activated cell sorting (FACS), we detected significant levels of TACI on 23 of 23 B-CLL samples and all normal B-cell samples tested (Figure 1A and data not shown). Additionally, binding of soluble BLyS, which binds TACI, BCMA, and BAFF-R, was detected on all B-CLL (23 of 23) as well as all normal B cells examined (Figure 1A and data not shown). Mature B cells have been reported to express both BCMA and BAFF-R, but because antibodies to either of these receptors are not yet available, we used RT-PCR to measure expression of both receptors. As shown in Figure 1B, 7 of 7 B-CLL samples expressed BAFF-R mRNA as did normal B cells (3 of 3 and data not shown). BCMA mRNA was expressed at low levels in normal B cells (3 of 3 and data not shown) and there was a variable pattern of expression in the B-CLL samples. Thus, similar to normal human B cells, B-CLL cells express BLyS receptors and have the ability to bind soluble BLyS.
BLyS protects CLL B cells from apoptosis Initially, it was proposed that BLyS functioned to enhance B-cell proliferation following anti-IgM or CD40 stimulation as well as immunoglobulin secretion.4,5 However, recent literature now suggests that the predominant function of BLyS is maintenance and survival of the B-cell population and attenuation of B-cell apoptosis.15 Because B-CLL cells express the receptors for BLyS and they are characterized by high resistance to apoptosis, we next wanted to determine the influence of BLyS on CLL B cells. Using caspase-3 activation and PARP degradation as hallmark indicators of apoptosis,32 we examined the effects of BLyS on CLL B-cell survival. Three B-CLL patient samples treated with BLyS for 18 hours had an average of a 55% reduction in the level of caspase-3 activity compared with untreated controls (Figure 2A). The caspase-3-specific inhibitor Ac-DEVD-CHO was used as a control to demonstrate the specificity of the assay. Cleavage of PARP was also assessed after treatment of B-CLL cells with BLyS. Similar to what was seen in the caspase-3 activity assays, there was a significant reduction in the level of PARP degradation in the presence of BLyS (Figure 2B). B-CLL cells were treated with 10 µM chlorambucil as a positive control for caspase-3 activation and PARP cleavage.33 These data clearly indicate that BLyS reduces the level of apoptosis and enhances survival of B-CLL B cells.
Expression of BLyS by CLL B cells Because of evidence in the literature demonstrating tumor cell autocrine expression of APRIL,12 we next wished to determine whether B-CLL cells could express BLyS or APRIL in an autocrine manner. To examine this possibility, we used RT-PCR to detect BLyS and APRIL mRNA in highly purified CD19+ peripheral blood B cells from B-CLL patients. Of great interest, BLyS mRNA was detected in 11 of 23 patients and APRIL mRNA was detected in 2 of 23 patients (Figure 3A). By contrast, normal CD19+ peripheral blood B cells failed to express either molecule (Figure 3B). Monocyte-enriched PBMCs were used as a positive control for BLyS and APRIL expression (Figure 3B).
To further confirm the RT-PCR data, we next analyzed B-CLL cells for
expression of BLyS by quantitative real-time PCR (Figure 4A). BLyS and cyclophilin-B were
amplified from 5 B-CLL patient samples, HL60 cells, which are known to
express BLyS,34 the Raji and HT2260 B-cell lymphoma lines,
and HeLa cells. The mRNA expression levels of cyclophilin-B and BLyS
are shown in Figure 4A. For analysis of BLyS expression, each sample
was normalized to cyclophilin-B and compared with the levels of BLyS
expression in HL60 cells (see "Materials and methods"). Relative to
HL60 cells, CLL B cells expressed lower levels (80%-95% less) of BLyS mRNA (Figure 4B). However, B-CLL cells clearly expressed BLyS, whereas
the 2 B-cell lymphoma lines and HeLa cells were found to be negative.
In addition to our PCR approach, we also examined B-CLL cells for cell
surface expression of BLyS (Figure 5).
B-CLL cells that were shown to express BLyS mRNA (Figure 3A) were also
found to express low levels of cell surface BLyS, whereas B-CLL cells that were negative by RT-PCR, were found to be negative for BLyS protein by FACS. As expected, normal B cells were deficient for BLyS
expression. Jurkat T cells stably expressing BLyS served as a positive
control. Work is currently underway to look for APRIL protein in B-CLL
cells by Western analysis, because it appears that APRIL may be
directly secreted without intermediate expression on the cell
surface.35 Taken together, these results suggest that B
CLL cells from a subset of patients aberrantly express BLyS and APRIL,
whereas these molecules were undetectable in normal B cells.
The data shown in Figure 2 demonstrated the ability of exogenous BLyS
to protect B-CLL cells from undergoing apoptosis. We next wished to
determine whether we could demonstrate a role for autocrine BLyS in the
maintenance and survival of CLL B-cells. To accomplish this, we used a
soluble decoy receptor approach. Thus, we examined the effects of
BCMA-Fc, which binds with high affinity to soluble BLyS and
APRIL,8,11,36 on B-CLL cell survival. Using
caspase-3 activation as a measurement of apoptosis we found that
incubation of CLL B cells, all of which expressed message for BLyS,
with BCMA-Fc resulted in enhanced caspase-3 activity in 4 of 5 patient
samples. Incubation of B-CLL cells with BCMA-Fc resulted in an average
of a 2.1-fold increase in caspase-3 activity compared with the negative
human IgG-Fc control (Figure 6). These
results suggest that treatment of CLL B cells with a blocking reagent
that binds soluble BLyS or APRIL may enhance apoptosis and decrease
cell survival.
The initial aim of our studies was to examine the role of BLyS and its receptors in B-CLL. Our results demonstrated that all B-CLL patients tested express TACI and BAFF-R, whereas only a subset express BCMA. Additionally, not only do B-CLL cells express the receptor for BLyS, they also have the ability to bind soluble BLyS. Finally, we also present evidence for the first time that B-CLL cells may also express BLyS and APRIL. The expression of TACI and BAFF-R by B-CLL cells was expected, because normal mature B cells express these receptors. However, the variable expression of BCMA by CLL B cells is intriguing and the significance of these finding is unclear at this time. However, it will be of utmost importance to more thoroughly examine the expression levels of TACI, BCMA, and BAFF-R in B-CLL in light of data suggesting that expression levels of BAFF-R highly influence the lifespan of B cells.37 A more definitive analysis of BCMA and BAFF-R protein expression by B-CLL cells is currently underway, however, this has been challenging because of a lack of reagents. Altered expression of BLyS receptors and ligands for these receptors may therefore contribute to the progressive accumulation of malignant B cells characteristic of B-CLL. Recent data support the notion that BLyS is essential to B-cell development and survival; however, the precise mechanism of BLyS action is less clearly understood. The ability of BLyS to bind 3 distinct receptors, BAFF-R, TACI, and BCMA, further complicates the picture. BAFF-R- and BLyS-deficient mice have similar phenotypes suggesting that BAFF-R is the predominant receptor for BLyS. However, BLyS clearly binds to and signals through TACI and BCMA. We provide evidence in this report that similar to normal B cells, B-CLL cells have enhanced cell survival in response to soluble BLyS. The ability of BLyS to down-regulate caspase-3 activity and PARP cleavage in B-CLL cells suggests a possible role for this protein in the maintenance and survival of malignant B-CLL cells. Currently, the molecular mechanisms underlying the pathogenesis of
B-CLL are poorly defined. However, there is a growing literature suggesting that CLL B cells have elevated levels of Bcl-238 as well as constitutive activation of NF- Accordingly, we investigated the possibility that deregulated expression of BLyS may contribute to the reduced levels of apoptosis found in B-CLL cells. In this report, we show for the first time that B-CLL cells from a subset of patients aberrantly express BLyS and APRIL, whereas these molecules were undetectable in normal B cells. The inability of normal B cells to produce BLyS is consistent with previous reports in the literature that have failed to detect BLyS expression in normal resting or activated B cells.4,34 Although there was no apparent association between Rai stage, CD38 expression, or mutation status and BLyS or APRIL expression (Table 1), this will need to be rigorously tested using larger numbers of B-CLL patient samples. Quantitative analysis of BLyS expression revealed that BLyS mRNA was found at low levels compared with HL60 cells and that it was virtually undetectable in B-cell lymphoma lines. Our conclusion that some B-CLL cells may express autocrine BLyS is further supported by our ability to detect low cell-surface expression of BLyS on CLL-B cells from some patients and the ability of a BCMA-Fc decoy receptor to increase apoptosis. Of interest, only 4 of 5 patient B-CLL samples cultured with BCMA-Fc displayed enhanced apoptosis. Although the significance of this observation remains unclear at this time, this result is consistent with the heterogeneity of this disease and may reflect heterogeneity in receptor expression levels or balance in expression between BAFF-R, TACI, and BCMA, or the inability or inefficiency of the decoy receptor to displace autocrine BLyS from cell-surface receptors. The mechanism by which some B-CLL cells acquire the ability to express BLyS and APRIL is currently unknown. However, evidence suggests that the locus for BLyS on chromosome 13q32-343 is amplified in human B-cell malignancies.42 Additionally, the ability of tumor cells to produce autocrine factors that support their growth is a common theme in many tumorigenesis models. Of interest, APRIL, another TNF superfamily member with homology to BLyS, is expressed by and enhances the in vivo growth rate of tumor cells.12 Although we have no evidence at this time that CLL-B cells express BLyS in vivo, we believe that it is unlikely that ex vivo manipulation of the CLL B cells would influence BLyS expression specifically in CLL B cells. Although CD19 microbead purification of CLL B cells may influence protein expression, because we purified normal and CLL B cells using identical methods, and because we did not find BLyS in either normal B cells or in a subset of B-CLL samples, we believe it is unlikely that our method of purification artificially induced BLyS expression. In addition, BLyS mRNA was expressed in patient samples not treated with CD19 microbeads that were more than 95% B-CLL cells (results not shown). Taken together, we believe our data strongly suggest that CLL B cells have the ability to synthesize BLyS. As noted previously, TACI is expressed on activated T cells.30 An additional biologic aspect relevant to our findings is the recent report that BAFF (BLyS) was able to induce T-cell activation and division under appropriate culture conditions.43 In addition, Wang and colleagues44 showed that TACI-ligand interactions were required for T-cell activation and collagen-induced arthritis in mice. Of interest, B-CLL patients not only have a clonal expansion of B cells at presentation, but also an expansion of polyclonal T cells.45 Thus, this raises the possibility that T-cell expansion may occur in part as a consequence of elevated BAFF/BLyS in the microenvironment. In this scenario of elevated BAFF/BLyS, however, it is difficult to explain the diminished normal B-cell compartment typically seen in B-CLL patients. One possible explanation for this observation is that the levels of BAFF-R, TACI, or BCMA expression on B-CLL cells may differ from normal B cells thereby providing a competitive advantage, that is, increased survival, to the leukemic population. Autocrine expression of BLyS by B-CLL cells has exciting implications and suggests the possibility that agents that inhibit BLyS binding to B-CLL cells may induce apoptosis. We have shown for the first time that B-CLL cells from a subset of patients aberrantly express BLyS and APRIL. Additionally, all B-CLL patients tested expressed TACI and BAFF-R, whereas only a minor subset expressed BCMA. Furthermore, we provide evidence that BLyS protects B-CLL cells from apoptosis and enhances cell survival, whereas the BCMA-Fc decoy receptor interrupted this autocrine loop resulting in enhanced cell death. We believe that these results are the first to show this unique autocrine pathway related to apoptosis in B-CLL. Importantly, our results also suggest that therapeutic agents that inhibit BLyS binding to B-CLL cells may have clinical efficacy.
We thank Ms Renee Tschumper and Nancy Bone for their help in determining B-CLL immunoglobulin mutation status and levels of CD38 expression.
Submitted February 21, 2002; accepted June 4, 2002.
Prepublished online as Blood First Edition Paper, June 14, 2002; DOI 10.1182/blood-2002-02-0558.
Supported by National Institutes of Health grant CA62228 (D.F.J.). A.J.N. was supported by a postdoctoral trainee award from the National Institutes of Health, grant CA09441.
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: Diane F. Jelinek, Department of Immunology, Mayo Clinic, 200 1st St SW, Rochester, MN 55905; e-mail: jelinek.diane{at}mayo.edu.
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  M. Gupta, S. R. Dillon, S. C. Ziesmer, A. L. Feldman, T. E. Witzig, S. M. Ansell, J. R. Cerhan, and A. J. Novak A proliferation-inducing ligand mediates follicular lymphoma B-cell proliferation and cyclin D1 expression through phosphatidylinositol 3-kinase-regulated mammalian target of rapamycin activation Blood, May 21, 2009; 113(21): 5206 - 5216. [Abstract] [Full Text] [PDF]
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 A. J. Novak, S. L. Slager, Z. S. Fredericksen, A. H. Wang, M. M. Manske, S. Ziesmer, M. Liebow, W. R. Macon, S. R. Dillon, T. E. Witzig, et al. Genetic Variation in B-Cell-Activating Factor Is Associated with an Increased Risk of Developing B-Cell Non-Hodgkin Lymphoma Cancer Res., May 15, 2009; 69(10): 4217 - 4224. [Abstract] [Full Text] [PDF]
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 L. Zhang, S. Zheng, H. Wu, Y. Wu, S. Liu, M. Fan, and J. Zhang Identification of BLyS (B Lymphocyte Stimulator), a Non-Myelin-Associated Protein, as a Functional Ligand for Nogo-66 Receptor J. Neurosci., May 13, 2009; 29(19): 6348 - 6352. [Abstract] [Full Text] [PDF]
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L. Fu, Y.-C. Lin-Lee, L. V. Pham, A. T. Tamayo, L. C. Yoshimura, and R. J. Ford BAFF-R promotes cell proliferation and survival through interaction with IKK{beta} and NF-{kappa}B/c-Rel in the nucleus of normal and neoplastic B-lymphoid cells Blood, May 7, 2009; 113(19): 4627 - 4636. [Abstract] [Full Text] [PDF]
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J. A. Burger, M. P. Quiroga, E. Hartmann, A. Burkle, W. G. Wierda, M. J. Keating, and A. Rosenwald High-level expression of the T-cell chemokines CCL3 and CCL4 by chronic lymphocytic leukemia B cells in nurselike cell cocultures and after BCR stimulation Blood, March 26, 2009; 113(13): 3050 - 3058. [Abstract] [Full Text] [PDF]
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S.-H. Kuo, P.-Y. Yeh, L.-T. Chen, M.-S. Wu, C.-W. Lin, K.-H. Yeh, Y.-S. Tzeng, J.-Y. Chen, P.-N. Hsu, J.-T. Lin, et al. Overexpression of B cell-activating factor of TNF family (BAFF) is associated with Helicobacter pylori-independent growth of gastric diffuse large B-cell lymphoma with histologic evidence of MALT lymphoma Blood, October 1, 2008; 112(7): 2927 - 2934. [Abstract] [Full Text] [PDF]
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 S. K. Chang, S. A. Mihalcik, and D. F. Jelinek B Lymphocyte Stimulator Regulates Adaptive Immune Responses by Directly Promoting Dendritic Cell Maturation J. Immunol., June 1, 2008; 180(11): 7394 - 7403. [Abstract] [Full Text] [PDF]
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 S. M. Ansell, T. E. Witzig, D. J. Inwards, L. F. Porrata, A. Ythier, L. Ferrande, I. Nestorov, T. DeVries, S. R. Dillon, D. Hausman, et al. Phase I Clinical Study of Atacicept in Patients with Relapsed and Refractory B-Cell Non-Hodgkin's Lymphoma Clin. Cancer Res., February 15, 2008; 14(4): 1105 - 1110. [Abstract] [Full Text] [PDF]
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S.-W. Kim, D. W. Oleksyn, R. M. Rossi, C. T. Jordan, I. Sanz, L. Chen, and J. Zhao Protein kinase C-associated kinase is required for NF-{kappa}B signaling and survival in diffuse large B-cell lymphoma cells Blood, February 1, 2008; 111(3): 1644 - 1653. [Abstract] [Full Text] [PDF]
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V. T. Chu, P. Enghard, G. Riemekasten, and C. Berek In Vitro and In Vivo Activation Induces BAFF and APRIL Expression in B Cells J. Immunol., November 1, 2007; 179(9): 5947 - 5957. [Abstract] [Full Text] [PDF]
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 A. Binard and P. Youinou BAFF, A newcomer to the lupus party Lupus, September 1, 2007; 16(9): 699 - 700. [PDF]
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 C. G. Mueller, C. Boix, W.-H. Kwan, C. Daussy, E. Fournier, W. H. Fridman, and T. J. Molina Critical role of monocytes to support normal B cell and diffuse large B cell lymphoma survival and proliferation J. Leukoc. Biol., September 1, 2007; 82(3): 567 - 575. [Abstract] [Full Text] [PDF]
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 L. Planelles, S. Castillo-Gutierrez, J. P. Medema, A. Morales-Luque, H. Merle-Beral, and M. Hahne APRIL but not BLyS serum levels are increased in chronic lymphocytic leukemia: prognostic relevance of APRIL for survival Haematologica, September 1, 2007; 92(9): 1284 - 1285. [Abstract] [Full Text] [PDF]
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J. R. Darce, B. K. Arendt, S. K. Chang, and D. F. Jelinek Divergent Effects of BAFF on Human Memory B Cell Differentiation into Ig-Secreting Cells J. Immunol., May 1, 2007; 178(9): 5612 - 5622. [Abstract] [Full Text] [PDF]
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Y. Oki, G. V. Georgakis, T.-S. Migone, L. W. Kwak, and A. Younes Prognostic significance of serum B-lymphocyte stimulator level in Hodgkin's lymphoma Haematologica, February 1, 2007; 92(2): 269 - 270. [Abstract] [Full Text] [PDF]
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 S. Kothlow, I. Morgenroth, Y. Graef, K. Schneider, I. Riehl, P. Staeheli, P. Schneider, and B. Kaspers Unique and conserved functions of B cell-activating factor of the TNF family (BAFF) in the chicken Int. Immunol., February 1, 2007; 19(2): 203 - 215. [Abstract] [Full Text] [PDF]
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 M.-A. Lyu, L. H. Cheung, W. N. Hittelman, J. W. Marks, R. C.T. Aguiar, and M. G. Rosenblum The rGel/BLyS fusion toxin specifically targets malignant B cells expressing the BLyS receptors BAFF-R, TACI, and BCMA Mol. Cancer Ther., February 1, 2007; 6(2): 460 - 470. [Abstract] [Full Text] [PDF]
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A. Chiu, W. Xu, B. He, S. R. Dillon, J. A. Gross, E. Sievers, X. Qiao, P. Santini, E. Hyjek, J.-w. Lee, et al. Hodgkin lymphoma cells express TACI and BCMA receptors and generate survival and proliferation signals in response to BAFF and APRIL Blood, January 15, 2007; 109(2): 729 - 739. [Abstract] [Full Text] [PDF]
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T. Endo, M. Nishio, T. Enzler, H. B. Cottam, T. Fukuda, D. F. James, M. Karin, and T. J. Kipps BAFF and APRIL support chronic lymphocytic leukemia B-cell survival through activation of the canonical NF-{kappa}B pathway Blood, January 15, 2007; 109(2): 703 - 710. [Abstract] [Full Text] [PDF]
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 D. Sene, N. Limal, P. Ghillani-Dalbin, D. Saadoun, J.-C. Piette, and P. Cacoub Hepatitis C virus-associated B-cell proliferation--the role of serum B lymphocyte stimulator (BLyS/BAFF) Rheumatology, January 1, 2007; 46(1): 65 - 69. [Abstract] [Full Text] [PDF]
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J. Schwaller, P. Schneider, P. Mhawech-Fauceglia, T. McKee, S. Myit, T. Matthes, J. Tschopp, O. Donze, F.-A. Le Gal, and B. Huard Neutrophil-derived APRIL concentrated in tumor lesions by proteoglycans correlates with human B-cell lymphoma aggressiveness Blood, January 1, 2007; 109(1): 331 - 338. [Abstract] [Full Text] [PDF]
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S. K. Chang, B. K. Arendt, J. R. Darce, X. Wu, and D. F. Jelinek A role for BLyS in the activation of innate immune cells Blood, October 15, 2006; 108(8): 2687 - 2694. [Abstract] [Full Text] [PDF]
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L. Fu, Y.-C. Lin-Lee, L. V. Pham, A. Tamayo, L. Yoshimura, and R. J. Ford Constitutive NF-{kappa}B and NFAT activation leads to stimulation of the BLyS survival pathway in aggressive B-cell lymphomas Blood, June 1, 2006; 107(11): 4540 - 4548. [Abstract] [Full Text] [PDF]
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D. Bischof, S. F. Elsawa, G. Mantchev, J. Yoon, G. E. Michels, A. Nilson, S. L. Sutor, J. L. Platt, S. M. Ansell, G. von Bulow, et al. Selective activation of TACI by syndecan-2 Blood, April 15, 2006; 107(8): 3235 - 3242. [Abstract] [Full Text] [PDF]
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S. F. Elsawa, A. J. Novak, D. M. Grote, S. C. Ziesmer, T. E. Witzig, R. A. Kyle, S. R. Dillon, B. Harder, J. A. Gross, and S. M. Ansell B-lymphocyte stimulator (BLyS) stimulates immunoglobulin production and malignant B-cell growth in Waldenstrom macroglobulinemia Blood, April 1, 2006; 107(7): 2882 - 2888. [Abstract] [Full Text] [PDF]
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K. N. Potter, C. I. Mockridge, L. Neville, I. Wheatley, M. Schenk, J. Orchard, A. S. Duncombe, G. Packham, and F. K. Stevenson Structural and Functional Features of the B-Cell Receptor in IgG-Positive Chronic Lymphocytic Leukemia. Clin. Cancer Res., March 15, 2006; 12(6): 1672 - 1679. [Abstract] [Full Text] [PDF]
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 A. J. Novak, D. M. Grote, S. C. Ziesmer, M. P. Kline, M. K. Manske, S. Slager, T. E. Witzig, T. Shanafelt, T. G. Call, N. E. Kay, et al. Elevated Serum B-Lymphocyte Stimulator Levels in Patients With Familial Lymphoproliferative Disorders J. Clin. Oncol., February 20, 2006; 24(6): 983 - 987. [Abstract] [Full Text] [PDF]
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J. S. Carew, S. T. Nawrocki, Y. V. Krupnik, K. Dunner Jr, D. J. McConkey, M. J. Keating, and P. Huang Targeting endoplasmic reticulum protein transport: a novel strategy to kill malignant B cells and overcome fludarabine resistance in CLL Blood, January 1, 2006; 107(1): 222 - 231. [Abstract] [Full Text] [PDF]
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M. Yang, H. Hase, D. Legarda-Addison, L. Varughese, B. Seed, and A. T. Ting B Cell Maturation Antigen, the Receptor for a Proliferation-Inducing Ligand and B Cell-Activating Factor of the TNF Family, Induces Antigen Presentation in B Cells J. Immunol., September 1, 2005; 175(5): 2814 - 2824. [Abstract] [Full Text] [PDF]
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J. Shaughnessy Jr APRIL showers cause CLL and myeloma to flower Blood, August 1, 2005; 106(3): 766 - 767. [Full Text] [PDF]
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M. Nishio, T. Endo, N. Tsukada, J. Ohata, S. Kitada, J. C. Reed, N. J. Zvaifler, and T. J. Kipps Nurselike cells express BAFF and APRIL, which can promote survival of chronic lymphocytic leukemia cells via a paracrine pathway distinct from that of SDF-1{alpha} Blood, August 1, 2005; 106(3): 1012 - 1020. [Abstract] [Full Text] [PDF]
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 T. Yamada, K. Zhang, A. Yamada, D. Zhu, and A. Saxon B Lymphocyte Stimulator Activates p38 Mitogen-Activated Protein Kinase in Human Ig Class Switch Recombination Am. J. Respir. Cell Mol. Biol., May 1, 2005; 32(5): 388 - 394. [Abstract] [Full Text] [PDF]
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 A Saxena, B Memauri, and W Hasegawa Initial diagnosis of small lymphocytic lymphoma in parotidectomy for Warthin tumour, a rare collision tumour J. Clin. Pathol., March 1, 2005; 58(3): 331 - 333. [Abstract] [Full Text] [PDF]
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C. A. Ogden, J. D. Pound, B. K. Batth, S. Owens, I. Johannessen, K. Wood, and C. D. Gregory Enhanced Apoptotic Cell Clearance Capacity and B Cell Survival Factor Production by IL-10-Activated Macrophages: Implications for Burkitt's Lymphoma J. Immunol., March 1, 2005; 174(5): 3015 - 3023. [Abstract] [Full Text] [PDF]
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P. Scapini, A. Carletto, B. Nardelli, F. Calzetti, V. Roschke, F. Merigo, N. Tamassia, S. Pieropan, D. Biasi, A. Sbarbati, et al. Proinflammatory mediators elicit secretion of the intracellular B-lymphocyte stimulator pool (BLyS) that is stored in activated neutrophils: implications for inflammatory diseases Blood, January 15, 2005; 105(2): 830 - 837. [Abstract] [Full Text] [PDF]
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A. J. Novak, D. M. Grote, M. Stenson, S. C. Ziesmer, T. E. Witzig, T. M. Habermann, B. Harder, K. M. Ristow, R. J. Bram, D. F. Jelinek, et al. Expression of BLyS and its receptors in B-cell non-Hodgkin lymphoma: correlation with disease activity and patient outcome Blood, October 15, 2004; 104(8): 2247 - 2253. [Abstract] [Full Text] [PDF]
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J. Moreaux, E. Legouffe, E. Jourdan, P. Quittet, T. Reme, C. Lugagne, P. Moine, J.-F. Rossi, B. Klein, and K. Tarte BAFF and APRIL protect myeloma cells from apoptosis induced by interleukin 6 deprivation and dexamethasone Blood, April 15, 2004; 103(8): 3148 - 3157. [Abstract] [Full Text] [PDF]
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B. Huard, L. Arlettaz, C. Ambrose, V. Kindler, D. Mauri, E. Roosnek, J. Tschopp, P. Schneider, and L. E. French BAFF production by antigen-presenting cells provides T cell co-stimulation Int. Immunol., March 1, 2004; 16(3): 467 - 475. [Abstract] [Full Text] [PDF]
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B. He, A. Chadburn, E. Jou, E. J. Schattner, D. M. Knowles, and A. Cerutti Lymphoma B Cells Evade Apoptosis through the TNF Family Members BAFF/BLyS and APRIL J. Immunol., March 1, 2004; 172(5): 3268 - 3279. [Abstract] [Full Text] [PDF]
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M. Batten, C. Fletcher, L. G. Ng, J. Groom, J. Wheway, Y. Laabi, X. Xin, P. Schneider, J. Tschopp, C. R. Mackay, et al. TNF Deficiency Fails to Protect BAFF Transgenic Mice against Autoimmunity and Reveals a Predisposition to B Cell Lymphoma J. Immunol., January 15, 2004; 172(2): 812 - 822. [Abstract] [Full Text] [PDF]
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J. C. Byrd BAFF and related proteins: a new therapeutic target for B-cell malignancies Blood, January 15, 2004; 103(2): 372 - 373. [Full Text] [PDF]
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C. Kern, J.-F. Cornuel, C. Billard, R. Tang, D. Rouillard, V. Stenou, T. Defrance, F. Ajchenbaum-Cymbalista, P.-Y. Simonin, S. Feldblum, et al. Involvement of BAFF and APRIL in the resistance to apoptosis of B-CLL through an autocrine pathway Blood, January 15, 2004; 103(2): 679 - 688. [Abstract] [Full Text] [PDF]
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A. J. Novak, J. R. Darce, B. K. Arendt, B. Harder, K. Henderson, W. Kindsvogel, J. A. Gross, P. R. Greipp, and D. F. Jelinek Expression of BCMA, TACI, and BAFF-R in multiple myeloma: a mechanism for growth and survival Blood, January 15, 2004; 103(2): 689 - 694. [Abstract] [Full Text] [PDF]
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 M. J. Keating, N. Chiorazzi, B. Messmer, R. N. Damle, S. L. Allen, K. R. Rai, M. Ferrarini, and T. J. Kipps Biology and Treatment of Chronic Lymphocytic Leukemia Hematology, January 1, 2003; 2003(1): 153 - 175. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
B (NF-
/
-actin
cDNAs were detected by PCR amplification with HotStarTaq
(Qiagen, Valencia, CA) in steps of 1 minute each at 94°C, 60°C and
72°C for 35 cycles, using primers previously described as being
specific for BLyS
values were collected for cyclophilin and BLyS during
log phase of the cycle. BLyS levels were normalized to cyclophilin for
each sample (
C
C
where
