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NEOPLASIA
From the Howard Hughes Medical Institute, Departments
of Biochemistry, Tumor Cell Biology, Immunology, and Experimental
Hematology, St. Jude Children's Research Hospital; and Department of
Biochemistry, University of Tennessee Medical School, Memphis, Memphis,
TN.
The cytokines interleukin 7 (IL-7) and interleukin 4 (IL-4)
regulate lymphoid differentiation and function and activate the transcription factor Stat5. Using mice deficient for the 2 highly related transcription factors, Stat5a and Stat5b
(Stat5a/b The cytokines interleukin 7 (IL-7) and interleukin
4 (IL-4) regulate important aspects of lymphoid development and
function and both activate the highly related transcription factors
Stat5a and Stat5b in B and T cells.1,2 In addition, strong
Stat5a/b activation was found in leukemias and has been hypothesized to be essential for the transformation process induced by derivatives of
the Abelson (abl) oncogene.3-5 In these studies
we addressed the importance of Stat5a/b activation by IL-7, IL-4, or
transforming protein tyrosine kinases using Stat5-deficient
mice.6
Disruption of the IL-7 gene and its high affinity receptor (IL-7R)
results in a dramatic loss of T- and B-cell precursors in the bone
marrow.7-9 Developing B cells undergo distinct
IL-7-dependent stages of expansion characterized by the expression of
specific surface markers (classified by Hardy et al10 and
Osmond et al11). Disruption of the IL-4 gene has shown its
importance in mature B-cell proliferation and immunoglobulin class
switching toward IgG1 and IgE as well as T helper 2 cell
differentiation and T-cell proliferation.12,13 IL-4
activates Stat5a, Stat5b, and Stat6 and we have previously shown that
Stat5-deficient T cells cannot proliferate in response to IL-2 or
IL-4.14-16 Stat6-deficient mice have a block in T helper 2 cell differentiation and immunoglobulin class switching toward IgE, but
B- and T-cell proliferation in response to IL-4 was not drastically
reduced and immunoglobulin class switching toward IgG1 was
unaltered.17 Lastly, IL-2 has been implicated in IgM
production through Stat5a/b.18,19 Therefore, we addressed
the question of whether B-cell development, proliferation, or
immunoglobulin production and class switching is impaired in Stat5a/b-deficient mice.
B-cell precursors are the target cells of the abl oncogene
and IL-7 can reconstitute multiple aspects of
v-abl-mediated signaling, among which the activation of
Stat5a/b is considered essential.20-23 The
abl gene was initially identified as a transforming gene
transduced into a Moloney leukemia virus (Ab-MuLV) and capable of
producing B-cell tumors in mice.24 Subsequently, it
was identified as a gene that fused with the break point cluster
gene (bcr) in leukemias containing a (9;22) translocation
termed the Philadelphia chromosome, which is associated with chronic
myeloid leukemia (CML), although it occurs also in acute
lymphocyte leukemia (ALL).25,26 In both cases, the
Abl protein is truncated and has increased protein kinase activity that
is required for its transforming activity. A variety of substrates
are phosphorylated in v-abl or bcr-abl transformed cells. Among the substrates that have been described as
strongly activated and potentially critical for abl-induced transformation are
Stat5a/b.3-5 20-23,27-30 Hence, we tested
this widely held hypothesis by assessing the ability of
v-abl or bcr-abl to induce disease in
Sta5a/b-deficient mice.
In vitro transformation assays
Determination of absolute B-cell numbers in the
peripheral blood
[3H]-thymidine incorporation assays To measure [3H]-thymidine incorporation into mature B cells, B220+ splenic B cells were purified by FACS sorting (all cells were Thy1.2 negative and CD19 positive; purity was greater than 99% for B220). The 1 × 105 cells per well were plated into 96-well round bottom plates and stimulated with either 100 ng/mL IL-4, 20 µg/mL -IgM, 1 µg/mL -CD40, 10 ng/mL LPS or
combinations of IL-4 with -IgM or -CD40. Forty-eight hours after
plating, [3H]-thymidine was added for the remaining
12 hours.
Immunizations and serum immunoglobulin detection To measure the T-cell-dependent response, mice were immunized with 100 µg ovalbumin (Sigma Chemical, St Louis, MO) in phosphate-buffered saline (PBS) and complete Freund's Adjuvant (Gibco BRL, Gaithersburg, MD) injected intraperitoneally. Serum samples were obtained 14 days after immunization and ovalbumin-specific responses were determined by enzyme-linked immunosorbent assay (ELISA), as described below. To measure the T-cell-independent immune response, mice were immunized with 50 µg NP-Ficoll (Biosearch Technologies, Novato, CA) in PBS injected intraperitoneally. Plasma samples were obtained 10 days after immunization and NP-specific antibody titers were determined by ELISA, as described below. Antibody titer was calculated as the fold increase compared with the average preimmunization titers of the wild-type control mice.For the ELISAs, mice were bled via the retro-orbital plexus, and serum
was collected after a 15-minute microcentrifuge spin at 4°C. Serum
was stored in aliquots at Bone marrow transduction and in vivo transformations A bicistronic retroviral vector using the murine stem cell virus (MSCV) long terminal repeats was used for expression of either bcr-abl p185 or p210. These constructs were generously provided by Dr Owen Witte. Viral supernatant was collected after transient transfection of 293T cells and ecotropic producer cell lines generated by repeated infection of GP+E86 cells with viral supernatant.31 Freshly isolated bone marrow cells were preactivated for 48 hours in medium containing IL-3 (25 ng/mL), IL-6 (50 ng/mL), and SCF (50 ng/mL) and were consequently cocultured on irradiated (1500 rads) ecotopic producer cell lines for 48 hours in the presence of 6 µg/mL polybrene. Lethally irradiated wild-type mice were reconstituted with the transduced bone marrow by tail vein injection (3 × 106 cells per mouse) and checked daily for the onset of disease. Diseased mice were killed. Blood samples and histopathologic sections were analyzed and used to diagnose the mice.Flow-cytometric analysis Single cell suspensions were preincubated withCD16/CD32
antibodies (Pharmingen) to prevent nonspecific Fc receptor-mediated binding. Thereafter, aliquots of 2 to 5 × 105 cells were
stained with monoclonal antibodies conjugated with fluorescent markers
(Pharmingen, San Diego, CA) and analyzed by FACSscan
(Becton-Dickinson, Franklin Lakes, NJ).
Protein analysis Analysis was performed as described previously.32 Briefly, cells were lysed in buffer containing 50 mmol/L HEPES buffer (pH 7.5), 0.1% Tween-20, 150 mmol/L NaCl, 1 mmol/L EDTA, 20 mmol/L -glycerophosphate, 0.1 mmol/L sodium vanadate, 1 mmol/L sodium fluoride, 10 µg/mL each aprotinin and leupeptin (both from Sigma Chemical), and 1 mmol/L PMSF. Protein concentrations were determined using a BCA-kit as recommended by the manufacturer (Pierce, Rockford, IL). To assess expression levels of the various forms of the Abl proteins, 100 µg total protein per sample was electrophoretically resolved on a 7.5% polyacrylamide gel containing SDS, and transferred onto Immobilon membranes. Membranes were probed with a rabbit polyclonal antiserum directed against Abl (Santa Cruz Biotechnology Inc, Santa Cruz, CA). Sites of antibody binding were detected using
protein A-conjugated horseradish peroxidase (EY Laboratories, San
Mateo, CA) with chemiluminescent detection (ECL detection kit,
Amersham, Arlington Heights, IL). Stat proteins were immunoprecipitated out of 600 µg of cell lysate using polyclonal antisera, resolved on a
7.5% gel, blotted, and probed with a monoclonal antibody directed
against phosphotyrosine (4G10, UBI) as described in detail in Moriggl
et al.16 Thereafter, blots were stripped and reprobed with
antisera directed against the individual Stat proteins (Transduction Laboratories, Santa Cruz, CA).
Northern blot analysis RNA of cell lines was isolated by RNAzol and separated on 1% agarose gels (20 µg per lane). A murine 0.2-kilobase (kb) BamHI-HindIII complementary DNA (cDNA) fragment for CIS, a 0.24-kb EcoRI fragment for OSM and a 1.2-kb EcoRI fragment for GAPDH, as loading control, were labeled by-[32P]dCTP using a
random labeling kit (Amersham). The membrane was hybridized using Rapid
hybridization solution (Amersham), followed by stringent washes (final
wash: 0.2 × SSC/0.1% SDS at 65°C).
Stat5a/b deficiency results in reduced numbers of pre- and pro- B cells In the initial studies of Stat5a/b-deficient mice,6 a significant reduction in IL-7-induced bone marrow colonies was observed. Because this population contains the cells that are transformed by abl, we further characterized these changes. As illustrated, the frequency of IL-7 colony-forming cells was significantly reduced (Figure 1A), as was the size of the colonies, consistent with our previous results. FACS analysis of the bone marrow demonstrated that there was an overall reduction of B cells by 70% (13% ± 1% CD19-positive cells vs 4% ± 3% in the Stat5a/b-deficient mice). The reduction is found preferentially within the B220-positive, CD43-intermediate and -low populations (Figure 2A,B). Together, the results suggest that, in the absence of Stat5a/b, there is a preferential loss of pre- and pro-B cells in the bone marrow.
In the peripheral blood of Stat5a/b-deficient mice, there is also a reduction in B220-positive B cells (Figure 2C) to levels approximately 6% that of control mice. However, the peripheral B cells that are present responded normally to stimulation with IL-4 and B-cell receptor engagement (Figure 2D), as well as to stimulation with anti-CD40 in the presence of IL-4. Moreover, B-cell numbers in the spleen are not reduced in Stat5a/b-deficient animals nor did we find any alterations or abnormalities in the distribution of mature versus immature B cells in the spleen (data not shown). Immunization of Stat5a/b-deficient mice with T-cell-dependent or -independent antigens resulted in comparable levels and isotypes of antibody production (Figure 2E,F). Together the results indicate that Stat5a/b-deficient mice have a preferential deficiency in the early IL-7-dependent phase of pre- and pro-B-cell development. Stat5a/b-deficient cells are susceptible to Ab-MuLV- and bcr-abl-induced transformation in vitro The transforming activity of Ab-MuLV and the bcr-abl oncogenes includes the ability to eliminate growth factor requirements of bone marrow progenitors. We therefore initially examined the response of bone marrow cells from Stat5a/b-deficient mice to be transformed to growth factor independence by Ab-MuLV and bcr-abl oncogenes. As illustrated in Figure 1B, the number, but not the size (data not shown) of growth factor-independent Ab-MuLV-induced bone marrow colony-forming cells was reduced in cells from Stat5a/b-deficient mice. As expected, immunocytochemical staining of the Ab-MuLV-transformed colonies revealed a B-lymphoid phenotype (data not shown). Therefore, we conclude that the number of Ab-MuLV targets are reduced consistent with the reduction in IL-7-responsive bone marrow cells. However, the cells that are present can be transformed in vitro comparable to wild-type cells.To explore the requirement of Stat5 proteins for bcr-abl
transformation, the ability to confer cytokine-independent growth of
Stat5a/b-deficient bone marrow cells was examined. Bone marrow cells
from Stat5a/b-deficient and wild-type mice were cocultured with cell
lines producing MSCV-based retroviruses expressing
bcr-abl-p210 or bcr-abl-p185 and GFP expressed
from an internal ribosome entry site (IRES). Three days after
infection, the cells were plated in methylcellulose and colony
formation assessed. Analysis of the transformed cells by Wright-Giemsa
staining and FACS analysis showed a myeloid phenotype for the
bcr-abl p210 transformed cells (data not shown). The cells
stained positive for the surface marker Mac1 and displayed myeloid
features independent of whether Stat5a/b were present. Transduction
with bcr-abl p185 resulted in myeloid as well as B lymphoid
transformants. In Stat5 (+/ Stat5a/b-deficient mice are susceptible to Ab-MuLV-induced tumors in vivo To assess the requirement for Stat5a/b for in vivo transformation, newborn mice were infected with replication-defective Ab-MuLV. Because the mice were obtained from crosses that use homozygous Stat5a/b-deficient males and heterozygous females, the heterozygous offspring served as controls. The majority (14 of 19; 74%) of the heterozygous animals developed B-cell lymphomas within 8 to 12 weeks, whereas the remainder survived longer than 6 months. Among the Stat5a/b-deficient animals, 7 of the 13 animals died from splenomegaly as a result of extramedullary hematopoiesis and anemia that is characteristically seen in nonmanipulated animals.16 Among the remaining 6 mice, 4 developed typical Ab-MuLV-induced pro-B-cell lymphomas at sites that were comparable to those seen with heterozygous control mice with a latency of 9 to 12 weeks (4 of 6; 67%). Histologically, the Stat5a/b-deficient tumors were identical to the tumors seen with control animals (Figure 3C,D). Also comparable to the tumors from control animals, the cells expressed B220, CD43, and CD19 (Figure 3E,F). In the 2 cases examined, 1 × 107 Stat5a/b-deficient tumor cells induced solid tumors in secondary SCID recipients (data not shown). Therefore, we conclude that Stat5a/b-deficient mice are susceptible to classical Ab-MuLV in vivo transformation.
Stat5a/b-deficient cells are susceptible to transformation by bcr-abl in vivo The in vivo transforming activities of bcr-abl-p210 and bcr-abl-p185 were examined by infecting bone marrow cells and reconstituting lethally irradiated mice.31 Infection of bone marrow cells from both wild-type and Stat5a/b-deficient mice resulted in the rapid onset of disease (18-24 days) with comparable pathologic alterations in all the recipients. The affected organs were lung (Figure 4A), spleen (Figure 4B), and liver (not shown). Histopathologic studies revealed extramedullary hematopoiesis in the spleen (Figure 4C), infiltration of tumor cells in the liver (Figure 4D), and lung (data not shown), resulting in the punctuated bleedings described by others.34,35 Analysis of peripheral blood from tumor-bearing mice was performed by blood smears and FACS (typical examples are illustrated in Figure 4E,F). The bcr-abl-p210 infection of wild-type bone marrow cells induced 8 myeloid lineage leukemias of the 9 leukemias phenotyped as defined by the expression of the myeloid lineage markers Mac1 and Gr1. In contrast, among the tumors derived from Stat5a/b-deficient cells that were phenotyped, 4 were of B-cell lineage, 2 had myeloid lineage markers, and 5 tumors contained both myeloid and B-cell lineage cells. The cell lineages of the tumors derived from bcr-abl-p185 were comparable between infection of wild-type or Stat5a/b-deficient bone marrow (summarized in Table 1). In both cases, B-cell lineage and myeloid lineage tumors or mixed tumors were observed.
For further analysis, cell lines were derived from tumors of wild- type
and Stat5a/b-deficient bone marrow (Table
2). With the exception of one myeloid
cell line from a Stat5a/b heterozygous animal expressing Mac1 and Gr1,
the cell lines that were established were of B-cell lineage and
expressed B220, CD43, and CD19. Cell lines were obtained from both
bcr-abl- and Ab-MuLV-infected wild-type and
Stat5a/b-deficient blood. As illustrated in Figure
5A, the various Abl proteins were highly
expressed in both control and Stat5a/b-deficient tumor cell lines. As
illustrated (Figure 5B), tumor cell lines from control bone marrow
expressed both the Stat5a/b regulated genes, CIS and
OSM, whereas the tumor cell lines from Stat5a/b-deficient
animals or bone marrow did not express either gene. This result
demonstrates that a redundant pathway, capable of replacing Stat5a/b
function in the regulation of these 2 genes, was not activated. Lastly,
tyrosine phosphorylation of various Stats is shown in Figure 5C.
Consistent with previous observations, Stat5a/b is tyrosine
phosphorylated in tumor cell lines from bcr-abl p185-transformed control bone marrow. This is not seen in tumor cell
lines from Stat5a/b-deficient marrow cells. No significant activation
of Stat1 or Stat3 was seen in any of the Stat5a/b-deficient tumor cell
lines.
IL-7 has been recognized as a major cytokine that stimulates
long-term proliferation and differentiation of precursor B
cells.1 Mice lacking IL-7 or the IL-7R Stat5a/b are activated by a broad variety of cytokines, among them IL-2, IL-4, and IL-7.2 We have recently shown that T cells lacking Stat5a/b have a proliferation defect in response to stimulation with either IL-2 or IL-4.15,16 In contrast, mature Stat5a/b-deficient B cells proliferate normally in response to IL-4 or when challenged with other B-cell mitogens (Figure 2D). So, despite its clear role in IL-4-induced T-cell proliferation, Stat5a/b are not required for the proliferation of mature B cells. Stat5a/b were also implicated in immunoglobulin class switching and IgM production,18,19,33 however, we failed to obtain evidence for this hypothesis. Stat5a/b-deficient mice responded with normal IgM and IgG1 levels when challenged with T-cell-dependent or T-cell-independent antigens (Figure 2E,F). Despite the large amount of literature,3-5,20-23,27-30 our results demonstrate that Stat5a/b are not essential for in vitro or in vivo transformation of B-cell precursors or myeloid cells by transforming derivatives of the abl proto-oncogene. It is well established that, depending on host and conditioning treatments, bcr-abl can induce T lineage disease,36,37 however, our conditions favored the B lymphoid and myeloid diseases. The myeloid disease can range from a myeloproliferative disease to the clonal expansion of myeloid lineage transformed cells. Further studies are necessary to explore the role of Stat5a/b in the full spectrum of diseases associated with bcr-abl and to explain the reduced frequency of pure myeloid leukemias arising in mice transplanted with Stat5a/b-deficient bone marrow. Our data prove, that Stat5a/b are not essential or necessary for Ab-MuLV- or bcr-abl-induced transformation. The possible basis for the shift of phenotype observed in mice transplanted with Stat5a/b-deficient bcr-abl-transformed bone marrow is not known. Because the colony assays did not reveal any alteration in myeloid colony formation, one may speculate that the reduction of myeloid disease in the animals is related to altered or disturbed cytokine production. It has been shown that bcr-abl transduction results in the production of several cytokines, eg, granulocyte-macrophage colony-stimulating factor (GM-CSF) and IL-3, and that cytokine production contributes to the myeloproliferative disease.38 The reduced myeloid disease might therefore also be related to decreased levels of cytokines, in particular the Stat5a/b target gene oncostatin M (OSM).39 OSM is shown to be expressed at high levels in the bone marrow.39 The lack of an effect on B lineage and myeloid transformation could be due to the utilization of pathways that can compensate for the absence of Stat5a/b. However, activation of Stat1 or Stat3 was not seen in the cell lines from Stat5a/b-deficient tumor cell lines. Moreover, although cell lines derived from wild-type mice expressed both the cytokine-inducible SH2-containing gene (CIS) and OSM, Stat5a/b target genes were not expressed in cell lines derived from tumors from Stat5a/b-deficient bone marrow cells or mice. Therefore, we have no evidence at this point that a redundant pathway is providing Stat5a/b functions. In summary, the results demonstrate that Stat5a/b are not critical substrates in bcr-abl- or abl-induced transformation of B cells or myeloid cells.
We thank M. Paktinat, S. Wingo, R. Cross, and R. Ashmun for technical assistance and J. Rehg, J. Downing, and M. Strain for help with the pathologic analysis of the diseased animals. We also thank Naomi Rosenberg and Owen Witte for the generous gift of A010 cells and the MSCV-210 and MSCV-p185 vectors, respectively.
Submitted January 27, 2000; accepted May 12, 2000.
Supported, in part, by Cancer Center CORE Grant (CA 21765) and ALSAC (American Lebanese Syrian Associated Charities).
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: James N. Ihle, Howard Hughes Medical Institute, St. Jude Children's Research Hospital, 332 N Lauderdale, Memphis, TN 38105; e-mail: james.ihle{at}stjude.org.
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X. Dai, Y. Chen, L. Di, A. Podd, G. Li, K. D. Bunting, L. Hennighausen, R. Wen, and D. Wang Stat5 Is Essential for Early B Cell Development but Not for B Cell Maturation and Function J. Immunol., July 15, 2007; 179(2): 1068 - 1079. [Abstract] [Full Text] [PDF]
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 R. A. Van Etten Oncogenic signaling: new insights and controversies from chronic myeloid leukemia J. Exp. Med., March 19, 2007; 204(3): 461 - 465. [Abstract] [Full Text] [PDF]
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 C.-H. Ting, C.-W. Lin, S.-L. Wen, H.-M. Hsieh-Li, and H. Li Stat5 constitutive activation rescues defects in spinal muscular atrophy Hum. Mol. Genet., March 1, 2007; 16(5): 499 - 514. [Abstract] [Full Text] [PDF]
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N. Harir, C. Pecquet, M. Kerenyi, K. Sonneck, B. Kovacic, R. Nyga, M. Brevet, I. Dhennin, V. Gouilleux-Gruart, H. Beug, et al. Constitutive activation of Stat5 promotes its cytoplasmic localization and association with PI3-kinase in myeloid leukemias Blood, February 15, 2007; 109(4): 1678 - 1686. [Abstract] [Full Text] [PDF]
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D. C. Otero, V. Poli, M. David, and R. C. Rickert Cutting Edge: Inherent and Acquired Resistance to Radiation-Induced Apoptosis in B Cells: A Pivotal Role for STAT3 J. Immunol., November 15, 2006; 177(10): 6593 - 6597. [Abstract] [Full Text] [PDF]
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W.-C. Chou, D. E. Levy, and C.-K. Lee STAT3 positively regulates an early step in B-cell development Blood, November 1, 2006; 108(9): 3005 - 3011. [Abstract] [Full Text] [PDF]
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D. K. Taylor, P. T. Walsh, D. F. LaRosa, J. Zhang, M. A. Burchill, M. A. Farrar, and L. A. Turka Constitutive Activation of STAT5 Supersedes the Requirement for Cytokine and TCR Engagement of CD4+ T Cells in Steady-State Homeostasis J. Immunol., August 15, 2006; 177(4): 2216 - 2223. [Abstract] [Full Text] [PDF]
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A. Hoelbl, B. Kovacic, M. A. Kerenyi, O. Simma, W. Warsch, Y. Cui, H. Beug, L. Hennighausen, R. Moriggl, and V. Sexl Clarifying the role of Stat5 in lymphoid development and Abelson-induced transformation Blood, June 15, 2006; 107(12): 4898 - 4906. [Abstract] [Full Text] [PDF]
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D. Ye, N. Wolff, L. Li, S. Zhang, and R. L. Ilaria Jr STAT5 signaling is required for the efficient induction and maintenance of CML in mice Blood, June 15, 2006; 107(12): 4917 - 4925. [Abstract] [Full Text] [PDF]
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 M. Nieborowska-Skorska, G. Hoser, L. Rink, M. Malecki, P. Kossev, M. A. Wasik, and T. Skorski Id1 transcription inhibitor-matrix metalloproteinase 9 axis enhances invasiveness of the breakpoint cluster region/abelson tyrosine kinase-transformed leukemia cells. Cancer Res., April 15, 2006; 66(8): 4108 - 4116. [Abstract] [Full Text] [PDF]
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 Z. Yao, Y. Cui, W. T. Watford, J. H. Bream, K. Yamaoka, B. D. Hissong, D. Li, S. K. Durum, Q. Jiang, A. Bhandoola, et al. Stat5a/b are essential for normal lymphoid development and differentiation PNAS, January 24, 2006; 103(4): 1000 - 1005. [Abstract] [Full Text] [PDF]
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M. Deininger, E. Buchdunger, and B. J. Druker The development of imatinib as a therapeutic agent for chronic myeloid leukemia Blood, April 1, 2005; 105(7): 2640 - 2653. [Abstract] [Full Text] [PDF]
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M. G. Kharas and D. A. Fruman ABL Oncogenes and Phosphoinositide 3-Kinase: Mechanism of Activation and Downstream Effectors Cancer Res., March 15, 2005; 65(6): 2047 - 2053. [Abstract] [Full Text] [PDF]
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S. Koschmieder, B. Gottgens, P. Zhang, J. Iwasaki-Arai, K. Akashi, J. L. Kutok, T. Dayaram, K. Geary, A. R. Green, D. G. Tenen, et al. Inducible chronic phase of myeloid leukemia with expansion of hematopoietic stem cells in a transgenic model of BCR-ABL leukemogenesis Blood, January 1, 2005; 105(1): 324 - 334. [Abstract] [Full Text] [PDF]
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J. Kang, B. DiBenedetto, K. Narayan, H. Zhao, S. D. Der, and C. A. Chambers STAT5 Is Required for Thymopoiesis in a Development Stage-Specific Manner J. Immunol., August 15, 2004; 173(4): 2307 - 2314. [Abstract] [Full Text] [PDF]
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B. Calabretta and D. Perrotti The biology of CML blast crisis Blood, June 1, 2004; 103(11): 4010 - 4022. [Abstract] [Full Text] [PDF]
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M. G. Kharas, J. A. Deane, S. Wong, K. R. O'Bosky, N. Rosenberg, O. N. Witte, and D. A. Fruman Phosphoinositide 3-kinase signaling is essential for ABL oncogene-mediated transformation of B-lineage cells Blood, June 1, 2004; 103(11): 4268 - 4275. [Abstract] [Full Text] [PDF]
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C. A. Goetz, I. R. Harmon, J. J. O'Neil, M. A. Burchill, and M. A. Farrar STAT5 Activation Underlies IL7 Receptor-Dependent B Cell Development J. Immunol., April 15, 2004; 172(8): 4770 - 4778. [Abstract] [Full Text] [PDF]
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 C. Heath and N. C. P. Cross Critical Role of STAT5 Activation in Transformation Mediated by ZNF198-FGFR1 J. Biol. Chem., February 20, 2004; 279(8): 6666 - 6673. [Abstract] [Full Text] [PDF]
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D. W. Sternberg and D. G. Gilliland The Role of Signal Transducer and Activator of Transcription Factors in Leukemogenesis J. Clin. Oncol., January 15, 2004; 22(2): 361 - 371. [Abstract] [Full Text] [PDF]
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A. V. Kazansky, D. M. Spencer, and N. M. Greenberg Activation of Signal Transducer and Activator of Transcription 5 is Required for Progression of Autochthonous Prostate Cancer: Evidence from the Transgenic Adenocarcinoma of the Mouse Prostate System Cancer Res., December 15, 2003; 63(24): 8757 - 8762. [Abstract] [Full Text] [PDF]
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M. A. Burchill, C. A. Goetz, M. Prlic, J. J. O'Neil, I. R. Harmon, S. J. Bensinger, L. A. Turka, P. Brennan, S. C. Jameson, and M. A. Farrar Distinct Effects of STAT5 Activation on CD4+ and CD8+ T Cell Homeostasis: Development of CD4+CD25+ Regulatory T Cells versus CD8+ Memory T Cells J. Immunol., December 1, 2003; 171(11): 5853 - 5864. [Abstract] [Full Text] [PDF]
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A. P. Szremska, L. Kenner, E. Weisz, R. G. Ott, E. Passegue, M. Artwohl, M. Freissmuth, R. Stoxreiter, H.-C. Theussl, S. B. Parzer, et al. JunB inhibits proliferation and transformation in B-lymphoid cells Blood, December 1, 2003; 102(12): 4159 - 4165. [Abstract] [Full Text] [PDF]
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J. W. Snow, N. Abraham, M. C. Ma, B. G. Herndier, A. W. Pastuszak, and M. A. Goldsmith Loss of Tolerance and Autoimmunity Affecting Multiple Organs in STAT5A/5B-Deficient Mice J. Immunol., November 15, 2003; 171(10): 5042 - 5050. [Abstract] [Full Text] [PDF]
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 M. W. N. Deininger and B. J. Druker Specific Targeted Therapy of Chronic Myelogenous Leukemia with Imatinib Pharmacol. Rev., September 1, 2003; 55(3): 401 - 423. [Abstract] [Full Text] [PDF]
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V. Sexl, B. Kovacic, R. Piekorz, R. Moriggl, D. Stoiber, A. Hoffmeyer, R. Liebminger, O. Kudlacek, E. Weisz, K. Rothammer, et al. Jak1 deficiency leads to enhanced Abelson-induced B-cell tumor formation Blood, June 15, 2003; 101(12): 4937 - 4943. [Abstract] [Full Text] [PDF]
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S. Wong, J. McLaughlin, D. Cheng, and O. N. Witte Cell context-specific effects of the BCR-ABL oncogene monitored in hematopoietic progenitors Blood, May 15, 2003; 101(10): 4088 - 4097. [Abstract] [Full Text] [PDF]
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 D. Warren, D. S. Griffin, C. Mainville, and N. Rosenberg The Extreme Carboxyl Terminus of v-Abl Is Required for Lymphoid Cell Transformation by Abelson Virus J. Virol., April 15, 2003; 77(8): 4617 - 4625. [Abstract] [Full Text] [PDF]
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M. Benekli, M. R. Baer, H. Baumann, and M. Wetzler Signal transducer and activator of transcription proteins in leukemias Blood, April 15, 2003; 101(8): 2940 - 2954. [Abstract] [Full Text] [PDF]
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H. L. Bradley, T. S. Hawley, and K. D. Bunting Cell intrinsic defects in cytokine responsiveness of STAT5-deficient hematopoietic stem cells Blood, December 1, 2002; 100(12): 3983 - 3989. [Abstract] [Full Text] [PDF]
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X. Jiang, E. Ng, C. Yip, W. Eisterer, Y. Chalandon, M. Stuible, A. Eaves, and C. J. Eaves Primitive interleukin 3 null hematopoietic cells transduced with BCR-ABL show accelerated loss after culture of factor-independence in vitro and leukemogenic activity in vivo Blood, November 15, 2002; 100(10): 3731 - 3740. [Abstract] [Full Text] [PDF]
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M. Nieborowska-Skorska, G. Hoser, P. Kossev, M. A. Wasik, and T. Skorski Complementary functions of the antiapoptotic protein A1 and serine/threonine kinase pim-1 in the BCR/ABL-mediated leukemogenesis Blood, May 29, 2002; 99(12): 4531 - 4539. [Abstract] [Full Text] [PDF]
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J. Dierov, Q. Xu, R. Dierova, and M. Carroll TEL/platelet-derived growth factor receptor beta activates phosphatidylinositol 3 (PI3) kinase and requires PI3 kinase to regulate the cell cycle Blood, March 1, 2002; 99(5): 1758 - 1765. [Abstract] [Full Text] [PDF]
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K. D. Bunting, H. L. Bradley, T. S. Hawley, R. Moriggl, B. P. Sorrentino, and J. N. Ihle Reduced lymphomyeloid repopulating activity from adult bone marrow and fetal liver of mice lacking expression of STAT5 Blood, January 15, 2002; 99(2): 479 - 487. [Abstract] [Full Text] [PDF]
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S. Ghaffari, C. Kitidis, M. D. Fleming, H. Neubauer, K. Pfeffer, and H. F. Lodish Erythropoiesis in the absence of janus-kinase 2: BCR-ABL induces red cell formation in JAK2-/- hematopoietic progenitors Blood, November 15, 2001; 98(10): 2948 - 2957. [Abstract] [Full Text] [PDF]
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 K. Miyoshi, J. M. Shillingford, G. H. Smith, S. L. Grimm, K.-U. Wagner, T. Oka, J. M. Rosen, G. W. Robinson, and L. Hennighausen Signal transducer and activator of transcription (Stat) 5 controls the proliferation and differentiation of mammary alveolar epithelium J. Cell Biol., November 12, 2001; 155(4): 531 - 542. [Abstract] [Full Text] [PDF]
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N. C. Wolff and R. L. Ilaria Jr Establishment of a murine model for therapy-treated chronic myelogenous leukemia using the tyrosine kinase inhibitor STI571 Blood, November 1, 2001; 98(9): 2808 - 2816. [Abstract] [Full Text] [PDF]
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V. M. Gelfanov, G. S. Burgess, S. Litz-Jackson, A. J. King, M. S. Marshall, H. Nakshatri, and H. S. Boswell Transformation of interleukin-3-dependent cells without participation of Stat5/bcl-xL: cooperation of akt with raf/erk leads to p65 nuclear factor {kappa}B-mediated antiapoptosis involving c-IAP2 Blood, October 15, 2001; 98(8): 2508 - 2517. [Abstract] [Full Text] [PDF]
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J. Frantsve, J. Schwaller, D. W. Sternberg, J. Kutok, and D. G. Gilliland Socs-1 Inhibits TEL-JAK2-Mediated Transformation of Hematopoietic Cells through Inhibition of JAK2 Kinase Activity and Induction of Proteasome-Mediated Degradation Mol. Cell. Biol., May 15, 2001; 21(10): 3547 - 3557. [Abstract] [Full Text]
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S. Mahajan, A. Vassilev, N. Sun, Z. Ozer, C. Mao, and F. M. Uckun Transcription Factor STAT5A Is a Substrate of Bruton's Tyrosine Kinase in B Cells J. Biol. Chem., August 10, 2001; 276(33): 31216 - 31228. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
/
) and (+/+) versus Stat5a/b (
) and in Stat5-deficient cells
(
) is shown. (E) Determination of the antibody response to the
T-cell-dependent antigen ovalbumin. Ovalbumin-specific serum IgM and
IgG1 titration curves are shown. Each dilution curve represents the
average of 5 mice. 
, heterozygous mice; 
, Stat5a/b-deficient
mice. (F) measurement of the serum IgM response against the
T-cell-independent antigen NP-Ficoll. Results are the average of 5 mice. Preimmunization titers, 
-chain.
J Biol Chem.
1998;47:31222-31229.