scholarly journals Expression of interleukin-4 receptors on early human B-lineage cells

Blood ◽  
1991 ◽  
Vol 78 (3) ◽  
pp. 703-710 ◽  
Author(s):  
CL Law ◽  
RJ Armitage ◽  
JG Villablanca ◽  
TW LeBien
Keyword(s):  
Bone Marrow ◽  
B Cell ◽  
Constitutive Expression ◽  
Lymphoid Cells ◽  
Interleukin 4 ◽  
B Cell Differentiation ◽  
Fluorescent Intensity ◽  
B Cell Growth Factor ◽  
B Lineage

Interleukin-4 (IL-4) regulates multiple stages of the antigen-dependent phase of B-cell development. However, its precise role in regulating B lymphopoiesis in bone marrow is not as well defined. We examined whether surface IgM- normal and leukemic human B-cell precursors (BCP) expressed IL-4 receptors using biotinylated IL-4. Constitutive expression of IL-4 receptors was detected on both normal and leukemic BCP. A higher percentage of normal BCP (82% +/- 15%) expressed IL-4 receptors compared with leukemic BCP (44% +/- 8%). Using mean fluorescent intensity as an indicator of receptor level on the IL-4 receptor positive cells, normal (91 +/- 41) and leukemic (44 +/- 37) BCP expressed comparable numbers of receptors. IL-4 induced the expression of CD23 on 30% of the leukemic BCP cases examined. IL-4 induced CD23 on surface IgM+ fetal bone marrow lymphoid cells but not on the surface IgM- normal BCP, despite the presence of detectable receptors on the surface IgM- cells. IL-4 did not stimulate proliferation of normal BCP, nor could it enhance the effect of recombinant IL-7 or low molecular weight B-cell growth factor. However, IL-4 increased the expression of surface IgM and surface Ig kappa on in vitro differentiated pre-B cells. Our collective results identify no role for IL-4 in the proliferation of normal or leukemic BCP, but identify a role in the enhancement of surface Ig expression during pre- B to B-cell differentiation.

scholarly journals Expression of interleukin-4 receptors on early human B-lineage cells

Blood ◽  
1991 ◽  
Vol 78 (3) ◽  
pp. 703-710 ◽  
Author(s):  
CL Law ◽  
RJ Armitage ◽  
JG Villablanca ◽  
TW LeBien
Keyword(s):  
Bone Marrow ◽  
B Cell ◽  
Constitutive Expression ◽  
Lymphoid Cells ◽  
Interleukin 4 ◽  
B Cell Differentiation ◽  
Fluorescent Intensity ◽  
B Cell Growth Factor ◽  
B Lineage

Abstract Interleukin-4 (IL-4) regulates multiple stages of the antigen-dependent phase of B-cell development. However, its precise role in regulating B lymphopoiesis in bone marrow is not as well defined. We examined whether surface IgM- normal and leukemic human B-cell precursors (BCP) expressed IL-4 receptors using biotinylated IL-4. Constitutive expression of IL-4 receptors was detected on both normal and leukemic BCP. A higher percentage of normal BCP (82% +/- 15%) expressed IL-4 receptors compared with leukemic BCP (44% +/- 8%). Using mean fluorescent intensity as an indicator of receptor level on the IL-4 receptor positive cells, normal (91 +/- 41) and leukemic (44 +/- 37) BCP expressed comparable numbers of receptors. IL-4 induced the expression of CD23 on 30% of the leukemic BCP cases examined. IL-4 induced CD23 on surface IgM+ fetal bone marrow lymphoid cells but not on the surface IgM- normal BCP, despite the presence of detectable receptors on the surface IgM- cells. IL-4 did not stimulate proliferation of normal BCP, nor could it enhance the effect of recombinant IL-7 or low molecular weight B-cell growth factor. However, IL-4 increased the expression of surface IgM and surface Ig kappa on in vitro differentiated pre-B cells. Our collective results identify no role for IL-4 in the proliferation of normal or leukemic BCP, but identify a role in the enhancement of surface Ig expression during pre- B to B-cell differentiation.

scholarly journals A Selective Culture System for Generating Terminal Deoxynucleotidyl Transferase-Positive Lymphoid CellsIn Vitro. V. Detection of Stage-Specific Pro-B-Cell Stimulating Activity in Medium Conditioned by Mouse Bone Marrow Stromal Cells

1993 ◽  
Vol 3 (3) ◽  
pp. 181-195 ◽  
Author(s):  
Sean D. Mckenna ◽  
Irving Goldschneider
Keyword(s):  
Bone Marrow ◽  
B Cell ◽  
Stromal Cells ◽  
Culture System ◽  
Lymphoid Cells ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Cell Phenotype ◽  
B Lineage

The selectivein vitrogeneration of rat, mouse, and human terminal deoxynucleotidyl transferase-positive (TdT+lymphoid cells in our long-term xenogeneic bone marrow (BM) culture system is characterized by physical interaction between the developing lymphocytes and mouse BM-adherent stromal cells and macrophages. In the present study, experiments in which micropor)us membrane culture inserts were inoculated with rat BM cells demonstrated that although the generation of primitive B-lineage lymphoid cells requires the presence of a mouse BM feeder layer, cognitive recognition events are not necessary. Similarly, cell-free (and serum-free) medium conditioned with mouse BM (but not thymus or spleen) adherent cells and stromal-cell lines therefrom supported the proliferation of early rat lymphoid cells in a dose-dependent manner. Double immunofluorescence for incorporated bromo-deoxyuridine (BrdU) and early B-lineage markers of rat BM lymphoid cells maintained in culture inserts or conditioned medium (CM), and studies of their in vitro andin vivodevelopmental potentials, indicated that the lymphoproliferative response resulted from the selective stimulation of lymphoid stem and/or progenitor cells. The most primitive of these target cells had a HIS24+HIS50-TdT-cμ-sIg-, pre-pro-B-cell phenotype. Whereas this subset normally constitutes less than 2% of B-lineage BM cellsin vivo, it comprises more than 25% of total lymphoid cellsin vitro. In addition, the number of TdT+cells, predominantly of the early pro-B-cell phenotype (HIS24+HIS50-TdT-cμ-sIg-), was increased approximately tenfold above input levels. Based on these and previous findings, a schematic model is proposed for the developmental pathway of early B-lineage cells in rat BM from the level of the committed (possibly common) lymphoid stem cell to that of the pre-B-cell.

scholarly journals Interleukin-4 induces programmed cell death (apoptosis) in cases of high-risk acute lymphoblastic leukemia

Blood ◽  
1994 ◽  
Vol 83 (7) ◽  
pp. 1731-1737 ◽  
Author(s):  
A Manabe ◽  
E Coustan-Smith ◽  
M Kumagai ◽  
FG Behm ◽  
SC Raimondi ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Death ◽  
Acute Lymphoblastic Leukemia ◽  
High Risk ◽  
Programmed Cell Death ◽  
B Cell ◽  
Lymphoblastic Leukemia ◽  
Fatal Outcome ◽  
Interleukin 4 ◽  
B Lineage

Abstract We investigated the effects of interleukin-4 (IL-4) on the survival of leukemic and normal B-cell progenitors cultured on bone marrow stroma. IL-4 (at 100 U/mL) was cytotoxic in 16 of 21 cases of B-lineage acute lymphoblastic leukemia, causing reductions in CD19+ cell numbers that ranged from 50% to greater than 99% (median 83.5%) of those in parallel cultures not exposed to the cytokine. All nine cases with the t(9;22)(q34;q11) or the t(4;11)(q21;q23), chromosomal features that are often associated with multidrug resistance and a fatal outcome, were susceptible to IL-4 toxicity. IL-4 cytotoxicity resulted from induction of programmed cell death (apoptosis); there was no evidence of cell killing mediated by T, natural killer, or stromal cells. IL-4 cytotoxicity extended to a proportion of normal B-cell progenitors. After 7 days of culture with IL-4 at 100 U/mL, fewer CD19+, CD34+ normal lymphoblasts (the most immature subset) survived: in five experiments the mean (+/- SEM) reduction in cell recoveries caused by IL-4 was 60.0% +/- 6.0%. By contrast, reductions in recovery of more differentiated bone marrow B cells (CD19+, CD34-, surface Ig+) were low (6.6% +/- 2.2%; P < .001 by t-test). Our findings indicate that IL-4 is cytotoxic for human B-cell precursors and support clinical testing of IL-4 in cases of high-risk lymphoblastic leukemia resistant to conventional therapy.

Molecular Cloning of DOCK10/Zizimin3, a Novel Cdc42/Rac-Interacting Protein Specifically Induced by IL4 in B Lymphocytes.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 939-939
Author(s):  
Estefania Yelo ◽  
Lourdes Gimeno ◽  
Maria Victoria Bernardo ◽  
Maria Juliana Majado ◽  
Maria Rocio Alvarez ◽  
...  
Keyword(s):  
Amino Acids ◽  
T Lymphocytes ◽  
Lymph Nodes ◽  
B Cell ◽  
B Lymphocytes ◽  
Expression System ◽  
Interleukin 4 ◽  
B Cell Differentiation ◽  
Nucleotide Exchange

Abstract Interleukin-4 (IL4) induces proliferation, differentiation and survival of B lymphocytes. IL4 protects CLL B cells from death by apoptosis. Gene expression analysis suggest that IL4 pathways are activated in CLL cells. We have identified DOCK10/Zizimin3 as an IL4-induced gene in CLL cells, and have obtained its full length sequence after cloning 1960 bp at its 5′ terminus by RACE-PCR. The human DOCK10/ZIZ3 sequence coded for a protein with 2180 amino acids and a predicted Mr of 250K. DOCK10/ZIZ3 shared homology with the other two members of the Zizimin family, and is the largest among them: DOCK9/ZIZ1 (2069 amino acids) and DOCK11/ZIZ2 (2073 amino acids) are 52% and 50% identical, respectively, to DOCK10/ZIZ3, and 58% identical between them. DOCK10 was predominantly expressed in hematopoietic tissues, particularly in peripheral blood (PB), but also in lymph nodes, thymus and spleen. Among the PB subpopulations, DOCK10 was expressed in B and T lymphocytes and, at lower levels, in monocytes. DOCK10 was also expressed in several non-hematopoietic tissues, most significantly in brain and kidney. Its homologue DOCK9, compared to DOCK10, was predominantly expressed in placenta, and less significantly in hematopoietic tissues, particularly in B lymphocytes and monocytes. DOCK11, like DOCK10, was predominantly expressed in PB. Compared to DOCK10, DOCK11 was expressed more prominently in placenta, thyroid and PB monocytes, and less significantly in brain and lymph nodes. Therefore, each of the Zizimin family members had a specific tissue distribution. Among the three genes, only DOCK10 was induced by IL4 in CLL cells in vitro. Induction of DOCK10 by IL4 was a common event in CLL, since it was observed in 10 out of 10 cases. IL4 also induced DOCK10 expression in normal PB B lymphocytes, suggesting that DOCK10 induction by IL4 in CLL cells may be normal, rather than pathological. Western blot analysis using a polyclonal antibody raised against a peptide which mapped at the N terminus of DOCK10, detected a band of the expected size of 250K. Interestingly, IL4 did not induce DOCK10 expression in CD4 or CD8 T lymphocytes in vitro. Expression of DOCK10 was also studied in 4 B-ALL, 2 T-ALL, and 1 T-CLL. DOCK10 neither was expressed at significant levels nor induced by IL4 in vitro in these patients, except for a weak induction in a common B-ALL case, suggesting that expression of DOCK10, and its induction with IL4, may be restricted to certain stages of B cell differentiation, and/or certain B cell malignancies. DOCK10 was distributed both in cytosolic and nuclear extracts of CLL cells, and IL4 increased its expression in both compartments. K562 clones stably transfected with DOCK10 using the inducible tet-off expression system showed significantly higher levels of DOCK10 in cytoplasm than in nucleus. Immunofluoresce analysis of HA-tagged DOCK10 K562 clones showed preferent staining of the cytoplasm, and dotted structures were frequently observed. GST-pulldown assays showed that DOCK10 bound to nucleotide-free (nf) Cdc42, but not to GTP- or GDP-loaded Cdc42. In addition, DOCK10 bound to nf Rac1, albeit with less affinity than to Cdc42. DOCK10 did not bind to RhoA. These results suggest that, like DOCK9 and DOCK11, DOCK10 may act as a novel Cdc42 guanine-nucleotide exchange factor (GEF) and, in addition, as a Rac1 GEF.

Trisomy of the Down Syndrome Critical Region Suppresses Precursor B-Cell Differentiation and Promotes B-Cell Transformation Associated with Altered Expression of Polycomb Repressor Complex 2 Targets

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 115-115
Author(s):  
Andrew A. Lane ◽  
Diederik van Bodegom ◽  
Bjoern Chapuy ◽  
Gabriela Alexe ◽  
Timothy J Sullivan ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Differentiation ◽  
B Cells ◽  
B Cell ◽  
Cell Transformation ◽  
B Cell Differentiation ◽  
Wild Type ◽  
Repressor Complex

Abstract Abstract 115 Extra copies of chromosome 21 (polysomy 21) is the most common somatic aneuploidy in B-cell acute lymphoblastic leukemia (B-ALL), including >90% of cases with high hyperdiploidy. In addition, children with Down syndrome (DS) have a 20-fold increased risk of developing B-ALL, of which ∼60% harbor CRLF2 rearrangements. To examine these associations within genetically defined models, we investigated B-lineage phenotypes in Ts1Rhr mice, which harbor triplication of 31 genes syntenic with the DS critical region (DSCR) on human chr.21. Murine pro-B cell (B220+CD43+) development proceeds sequentially through “Hardy fractions” defined by cell surface phenotype: A (CD24−BP-1−), B (CD24+BP-1−) and then C (CD24+BP-1+). Compared with otherwise isogenic wild-type littermates, Ts1Rhr bone marrow harbored decreased percentages of Hardy fraction B and C cells, indicating that DSCR triplication is sufficient to disrupt the Hardy A-to-B transition. Of note, the same phenotype was reported in human DS fetal liver B-cells, which have a block between the pre-pro- and pro-B cell stages (analogous to Hardy A-to-B). To determine whether DSCR triplication affects B-cell proliferation in vitro, we analyzed colony formation and serial replating in methylcellulose cultures. Ts1Rhr bone marrow (B6/FVB background) formed 2–3-fold more B-cell colonies in early passages compared to bone marrow from wild-type littermates. While wild-type B-cells could not serially replate beyond 4 passages, Ts1Rhr B-cells displayed indefinite serial replating (>10 passages). Ts1Rhr mice do not spontaneously develop leukemia, so we utilized two mouse models to determine whether DSCR triplication cooperates with leukemogenic oncogenes in vivo. First, we generated Eμ-CRLF2 F232C mice, which express the constitutively active CRLF2 mutant solely within B-cells. Like Ts1Rhr B-cells, (but not CRLF2 F232C B-cells) Ts1Rhr/CRLF2 F232C cells had indefinite serial replating potential. In contrast with Ts1Rhr B-cells, Ts1Rhr/CRLF2 F232C B-cells also engrafted into NOD.Scid.IL2Rγ−/− mice and caused fatal and serially transplantable B-ALL. Second, we retrovirally transduced BCR-ABL1 into unselected bone marrow from wild-type and Ts1Rhr mice and transplanted into irradiated wild-type recipients. Transplantation of transduced Ts1Rhr cells (106, 105, or 104) caused fatal B-ALL in recipient mice with shorter latency and increased penetrance compared to recipients of the same number of transduced wild-type cells. By Poisson calculation, the number of B-ALL initiating cells in transduced Ts1Rhr bone marrow was ∼4-fold higher than in wild-type animals (1:60 vs 1:244, P=0.0107). Strikingly, transplantation of individual Hardy A, B, and C fractions after sorting and BCR-ABL1 transduction demonstrated that the increased leukemia-initiating capacity almost completely resides in the Ts1Rhr Hardy B fraction; i.e., the same subset suppressed during Ts1Rhr B-cell differentiation. To define transcriptional determinants of these phenotypes, we performed RNAseq of Ts1Rhr and wild-type B cells in methylcellulose culture (n=3 biologic replicates per genotype). As expected, Ts1Rhr colonies had ∼1.5-fold higher RNA abundance of expressed DSCR genes. We defined a Ts1Rhr signature of the top 200 genes (false discovery rate (FDR) <0.25) differentially expressed compared with wild-type cells. Importantly, this Ts1Rhr signature was significantly enriched (P=0.02) in a published gene expression dataset of DS-ALL compared with non-DS-ALL (Hertzberg et al., Blood 2009). Query of >2,300 signatures in the Molecular Signatures Database (MSigDB) C2 Chemical and Genetic Perturbations with the Ts1Rhr signature identified enrichment in multiple gene sets of polycomb repressor complex (PRC2) targets and H3K27 trimethylation. Most notably, SUZ12 targets within human embryonic stem cells were more highly expressed in Ts1Rhr cells (P=1.2×10−6, FDR=0.003) and the same SUZ12 signature was enriched in patients with DS-ALL compared to non-DS-ALL (P=0.007). In summary, DSCR triplication directly suppresses precursor B-cell differentiation and promotes B-cell transformation both in vitro and by cooperating with proliferative alterations such as CRLF2 activation and BCR-ABL1 in vivo. Pharmacologic modulation of H3K27me3 effectors may overcome the pro-leukemogenic effects of polysomy 21. Disclosures: No relevant conflicts of interest to declare.

The development of functional B lymphocytes in conditional PU.1 knock-out mice

Blood ◽  
2005 ◽  
Vol 106 (6) ◽  
pp. 2083-2090 ◽  
Author(s):  
Matthew Polli ◽  
Aleksandar Dakic ◽  
Amanda Light ◽  
Li Wu ◽  
David M. Tarlinton ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
B Lymphocytes ◽  
Fetal Liver ◽  
Cell Receptor ◽  
Interleukin 4 ◽  
Knock Out ◽  
B Lineage ◽  
And Function

Abstract An abundance of research has entrenched the view that the Ets domain containing transcription factor PU.1 is fundamental to the development and function of B lymphocytes. In this study, we have made use of a conditional PU.1 allele to test this notion. Complete deletion of PU.1 resulted in the loss of B cells and all other lineage-positive cells in the fetal liver and death between E18.5 and birth; however, specific deletion of PU.1 in the B lineage had no effect on B-cell development. Furthermore, deletion of PU.1 in B cells did not compromise their ability to establish and maintain an immune response. An increased level of apoptosis was observed in vitro upon B-cell receptor (BCR) cross-linking; however, this was partially rescued by interleukin-4 (IL-4). These findings suggest that PU.1 is not essential for the development of functional B lymphocytes beyond the pre-B stage. (Blood. 2005;106:2083-2090)

scholarly journals Immunoglobulin recombinase gene activity is modulated reciprocally by interleukin 7 and CD19 in B cell progenitors.

1995 ◽  
Vol 182 (4) ◽  
pp. 973-982 ◽  
Author(s):  
L G Billips ◽  
C A Nuñez ◽  
F E Bertrand ◽  
A K Stankovic ◽  
G L Gartland ◽  
...  
Keyword(s):  
Bone Marrow ◽  
B Cell ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Immunoglobulin Gene ◽  
B Cell Differentiation ◽  
Interleukin 7 ◽  
Cross Linkage ◽  
Recombinase Gene ◽  
Rag 1 ◽  
B Lineage

Bone marrow stromal cells promote B cell development involving recombinase gene-directed rearrangement of the immunoglobulin genes. We observed that the stromal cell-derived cytokine interleukin 7 (IL-7) enhances the expression of CD19 molecules on progenitor B-lineage cells in human bone marrow samples and downregulates the expression of terminal deoxynucleotidyl transferase (TdT) and the recombinase-activating genes RAG-1 and RAG-2. Initiation of the TdT downregulation on the first day of treatment, CD19 upregulation during the second day, and RAG-1 and RAG-2 downmodulation during the third day implied a cascade of IL-7 effects. While CD19 ligation by divalent antibodies had no direct effect on TdT or RAG gene expression, CD19 cross-linkage complete blocked the IL-7 downregulation of RAG expression without affecting the earlier TdT response. These results suggest that signals generated through CD19 and the IL-7 receptor could modulate immunoglobulin gene rearrangement and repertoire diversification during the early stages of B cell differentiation.

scholarly journals Oncogene cooperation in lymphocyte transformation: malignant conversion of E mu-myc transgenic pre-B cells in vitro is enhanced by v-H-ras or v-raf but not v-abl.

1989 ◽  
Vol 9 (1) ◽  
pp. 67-73 ◽  
Author(s):  
W S Alexander ◽  
J M Adams ◽  
S Cory
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
Bone Marrow Cells ◽  
Lymphocyte Transformation ◽  
Lymphoid Cells ◽  
B Lineage ◽  
B Lymphoid ◽  
Marrow Cells ◽  
B Lineage Cells

Although transgenic mice bearing a c-myc gene controlled by the immunoglobulin heavy-chain enhancer (E mu) eventually develop B-lymphoid tumors, B-lineage cells from preneoplastic bone marrow express the transgene but do not grow autonomously or produce tumors in mice. To determine whether other oncogenes can cooperate with myc to transform B-lineage cells, we compared the in vitro growth and tumorigenicity of normal and E mu-myc bone marrow cells infected with retroviruses bearing the v-H-ras, v-raf, or v-abl oncogene. The v-H-ras and v-raf viruses both generated a rapid polyclonal expansion of E mu-myc pre-B bone marrow cells in liquid culture and 10- to 100-fold more pre-B lymphoid colonies than normal in soft agar. The infected transgenic cells were autonomous, cloned efficiently in agar, and grew as tumors in nude mice. While many pre-B cells from normal marrow could also be induced to proliferate by the v-raf virus, these cells required a stromal feeder layer, did not clone in agar, and were not malignant. Most normal cells stimulated to grow by v-H-ras also cloned poorly in agar, and only rare cells were tumorigenic. With the v-abl virus, no more cells were transformed from E mu-myc than normal marrow and the proportion of tumorigenic pre-B clones was not elevated. These results suggest that both v-H-ras and v-raf, but apparently not v-abl, collaborate with constitutive myc expression to promote autonomous proliferation and tumorigenicity of pre-B lymphoid cells.

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