The expression of Vpre-B/λ5 surrogate light chain in early bone marrow precursor B cells of normal and B cell-deficient mutant mice

Cell ◽  
1994 ◽  
Vol 77 (1) ◽  
pp. 133-143 ◽  
Author(s):  
Hajime Karasuyama ◽  
Antonius Rolink ◽  
Yoichi Shinkal ◽  
Faith Young ◽  
Frederick W. Alt ◽  
...  
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
B Cell ◽  
Light Chain ◽  
Mutant Mice ◽  
Deficient Mutant ◽  
Surrogate Light Chain ◽  
Bone Marrow Precursor ◽  
Precursor B Cells

Interleukin-3 and interleukin-7 are alternative growth factors for the same B-cell precursors in the mouse

Blood ◽  
1995 ◽  
Vol 85 (8) ◽  
pp. 2045-2051 ◽  
Author(s):  
TH Winkler ◽  
F Melchers ◽  
AG Rolink
Keyword(s):  
Bone Marrow ◽  
Growth Factors ◽  
B Cells ◽  
B Cell ◽  
Light Chain ◽  
Stromal Cells ◽  
Fetal Liver ◽  
Surrogate Light Chain ◽  
Interleukin 7 ◽  
Precursor B Cells

Clones and lines of precursor (pre) B cells can be established by limiting dilutions of unseparated cell suspensions of fetal liver or bone marrow on stromal cells in the presence of interleukin (IL)-7. When IL-3 is used instead of IL-7, cultures are regularly overgrown by different precursor cells of the myeloid lineage, as well as by adherent cells that inhibit pre-B-cell expansion. However, in the presence of either IL-7 or IL-3, clones of pre-B cells can be established on stroma cells at frequencies near one in one when the cultures are initiated with cell sorter purified CD45RO (B220)+/c-kit+ fetal liver or bone marrow derived pre-B cells. Clones grown on stromal cells in the presence of IL-7 can be regrown in IL-3, and vice versa. Pre-B cells that proliferate on stromal cells in the presence of IL-7 or IL-3 have the same phenotype, ie, are B220+ c-kit+, CD43+, and surrogate light chain+. Removal of the growth factors (IL-7, respectively IL-3) from the cultures results in differentiation to surface immunoglobulin (slg) positive, c-kit-, CD43-, surrogate light chain-B cells, a fraction of which is lipopolysaccharide (LPS) responsive as shown by IgM secretion. These results show that IL-7 and IL-3 stimulate largely overlapping populations of precursor B cells from bone marrow to proliferate for long periods of time in the presence of stromal cells. Thus, IL-7 and IL-3 are alternative growth factors for the same pre-BI cell.

Precursor B cells of mouse bone marrow express two different complexes with the surrogate light chain on the surface

1995 ◽  
Vol 25 (2) ◽  
pp. 446-450 ◽  
Author(s):  
Thomas H. Winkler ◽  
Antonius Rolink ◽  
Fritz Melchers ◽  
Hajime Karasuyama
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
Light Chain ◽  
Mouse Bone Marrow ◽  
Surrogate Light Chain ◽  
Precursor B Cells

scholarly journals B lymphocytes express and lose syndecan at specific stages of differentiation.

1989 ◽  
Vol 1 (1) ◽  
pp. 27-35 ◽  
Author(s):  
R D Sanderson ◽  
P Lalor ◽  
M Bernfield
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
B Cell ◽  
B Lymphocytes ◽  
B Lymphocyte ◽  
Plasma Cells ◽  
Receptor Expression ◽  
Cell Stage ◽  
Precursor B Cells

Lymphopoietic cells require interactions with bone marrow stroma for normal maturation and show changes in adhesion to matrix during their differentiation. Syndecan, a heparan sulfate-rich integral membrane proteoglycan, functions as a matrix receptor by binding cells to interstitial collagens, fibronectin, and thrombospondin. Therefore, we asked whether syndecan was present on the surface of lymphopoietic cells. In bone marrow, we find syndecan only on precursor B cells. Expression changes with pre-B cell maturation in the marrow and with B-lymphocyte differentiation to plasma cells in interstitial matrices. Syndecan on B cell precursors is more heterogeneous and slightly larger than on plasma cells. Syndecan 1) is lost immediately before maturation and release of B lymphocytes into the circulation, 2) is absent on circulating and peripheral B lymphocytes, and 3) is reexpressed upon their differentiation into immobilized plasma cells. Thus, syndecan is expressed only when and where B lymphocytes associate with extracellular matrix. These results indicate that B cells differentiating in vivo alter their matrix receptor expression and suggest a role for syndecan in B cell stage-specific adhesion.

Impaired Precursor B-Cell Maturation/Production In The Bone Marrow: Association With Molecular and Immunophenotypic Markers Of Myeloid Dysplasia In Suspected Low-Grade MDS

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 4943-4943
Author(s):  
Charles Repetti ◽  
Hsueh-Hua Chen ◽  
Yongbao Wang ◽  
Vanessa A Jones ◽  
Albert K Ho ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
B Cells ◽  
B Cell ◽  
Somatic Mutation ◽  
Conflicts Of Interest ◽  
Statistical Significance ◽  
Sequence Variant ◽  
Low Grade ◽  
Precursor B Cells

Abstract Rationale Myelodysplastic syndromes (MDS) are clonal stem cell disorders that disrupt orderly maturation of multiple hematopoietic lineages. Several studies have suggested that maturation of precursor B cells (hematogones) is also abnormal in MDS. As a result, the presence of normal numbers or increased precursor B cells in bone marrow (BM) is frequently used as a diagnostic feature arguing against a diagnosis of MDS. We compared the presence of myeloid-associated gene mutations and myeloid maturation abnormalities with qualitative and quantitative precursor B cell findings in BM samples submitted for workup of cytopenias or MDS. Methods Seventeen BM aspirate samples with <5% blasts submitted for cytopenia or MDS evaluation were compared with 10 samples having 5% or more blasts and changes diagnostic of MDS or AML. Mutation analysis was performed on genomic DNA using a targeted exome sequencing assay. This assay employs a TruSeq custom amplicon design on the MiSeq platform (Illumina, San Diego, CA). The assay covers the commonly mutated areas of 19 myeloid-associated genes. Somatic mutation status was assigned based on mutation levels, previous association with myeloid neoplasia, and no prior identification in public or internal databases as a normal sequence variant. Flow cytometry using 6-color (CD19/CD34) and 8-color (CD19/10) formats was used to assess lymphoblasts; CD34/13 was used to assess myeloblasts; and CD11b, CD13, CD16, and CD38 were used to assess abnormalities in myelopoiesis. Results  Among the 17 BM samples submitted for cytopenia or MDS evaluation that had <5% blasts, 7 (41%) had immunophenotypic myeloid maturation abnormalities. Ten (59%) of the 17 cases had at least one myeloid-associated somatic mutation, with TET2 and ASXL1being the most commonly mutated genes. The ratio of myeloblasts to B-lymphoblasts, calculated using either CD10 or CD19, was >10:1 in 10/17 (59%) cases. Nine of the 17 (53%) cases had virtually no precursor B cells detected. Discrete abnormalities in more mature myeloid forms were seen in 7/10 (70%) cases with low numbers of B-lymphoblasts but in none of the 7 cases with significant numbers of B-lymphoblasts. MDS-associated mutations were more common in cases with rare B-lymphoblasts (7/9) than in those with higher percentages of precursor B cells (3/8), but the difference did not reach statistical significance (P = 0.15).  Genes mutated in the group with B-lymphoblasts present included ASXL1 (3 cases), DNMT3A (2), TET2 (1) and TP53 (2). Two of these mutated cases presented with isolated thrombocytopenia. By comparison, myeloblast/lymphoblast ratios were >50:1 in all 10 unequivocal MDS/AML samples (>5% blasts); 8 (80%) of these cases had MDS-associated mutations, and 4 (50%) had mutations in multiple genes. Conclusions Decreases in BM precursor B cells in cases of possible low-grade MDS were usually, but not always, associated with the presence of MDS-associated mutations. However, cases with normal or increased precursor B cell numbers also showed MDS-associated mutations although immunophenotypic evidence of myeloid maturation abnormalities was not seen in this group. The identification of a subgroup of cytopenic patients with likely pathogenic mutations in bone marrow precursors but minimal phenotypic evidence of myeloid dysplasia may indicate clonal abnormalities primarily located outside the granulocyte or common stem precursor populations, e.g. restricted to the megakaryocytic lineage. Therefore, the presence of intact precursor lymphoblast and myeloid maturation by higher-dimensional flow cytometry as a primary criterion to argue against a diagnosis of low-grade MDS needs further evaluation, especially when granulocytopenia is absent. Disclosures: No relevant conflicts of interest to declare.

scholarly journals In aged mice, low surrogate light chain promotes pro‐ B ‐cell apoptotic resistance, compromises the P re BCR checkpoint, and favors generation of autoreactive, phosphorylcholine‐specific B cells

Aging Cell ◽  
2015 ◽  
Vol 14 (3) ◽  
pp. 382-390 ◽  
Author(s):  
Michelle Ratliff ◽  
Sarah Alter ◽  
Kelly McAvoy ◽  
Daniela Frasca ◽  
Jacqueline A. Wright ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
Light Chain ◽  
Aged Mice ◽  
Surrogate Light Chain

Quantitative analysis of clonal bone marrow CD19+ B cells: Use of B cell lineage trees to delineate their role in the pathogenesis of light chain amyloidosis

2006 ◽  
Vol 120 (1) ◽  
pp. 106-120 ◽  
Author(s):  
Michelle K. Manske ◽  
Neta S. Zuckerman ◽  
Michael M. Timm ◽  
Stephanie Maiden ◽  
Hanna Edelman ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Quantitative Analysis ◽  
B Cells ◽  
B Cell ◽  
Light Chain ◽  
Cell Lineage ◽  
Light Chain Amyloidosis ◽  
Lineage Trees

Pre-B cell receptor expression in B-lineage acute lymphoblastic leukemia: A report from the Children’s Oncology Group.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. e19006-e19006
Author(s):  
Stuart S. Winter ◽  
Amanda McCaustland ◽  
No'eau Simeona ◽  
Andrew J. Carroll ◽  
Nyla A. Heerema ◽  
...  
Keyword(s):  
Gene Expression ◽  
B Cells ◽  
B Cell ◽  
Light Chain ◽  
Lymphoblastic Leukemia ◽  
Surface Expression ◽  
Oncology Group ◽  
Surrogate Light Chain ◽  
Cell Ablation ◽  
B Lineage

e19006 Background: The surface expression of mature B-cell markers have led to the development of immunotherapies against B-lineage lymphoblastic leukemia/lymphoma (B-ALL/B-LLy). Relapsing clones that have altered surface antigen expression are common means of treatment failure with immunotherapies. The elimination of the pan-B cell repertoire by current B-cell immunotherapies contributes to immune-compromise. A promising target is the pre-BCR surrogate light chain, comprised of the VpreB1 (CD179a) and Lamda5 (CD179b) subunits. Surrogate light chain is expressed on pro- and pre-B cells where it governs preBCR-mediated autonomous survival during B-cell maturation. Gene expression analyses have shown that CD179a is expressed in a sub-set of 10 to 15% of B-ALL cases. Because immunotherapies targeted to restricted stages of B-cell development may overcome the limitations of pan B-cell ablation, we tested the hypothesis that CD179a is more commonly expressed on B-lymphoblasts than previously thought. Methods: Utilizing an annotated set of 36 standard (AALL0331) and high-risk (AALL0232) B-ALL cases accrued to Children’s Oncology Group AALL03B1, we adapted the COG minimal residual disease (MRD) flow panel to include two additional PE- and FITC-conjugated mAbs against CD179a (Biolegend and i2Pharma). We assessed CD179a expression in 16 cases for which we had Day 28 end-induction samples, pre-selected to have ≥1% MRD, as determined by the COG Reference laboratories. Cases with ≥20% CD179a surface expression were determined to be positive for statistical comparisons. All analyses were performed on a 6-color Becton-Dickinson flow cytometer in a CLIA/CAP certified laboratory. Results: Thirty-four cases were arrested at the CD10-positive pre-B stage, and two cases at the CD10-negative pro-B stage. One or both mAbs showed that CD179a was present in ≥20% of the B-lymphoblast population, ranging from 20.2% to 90.6% for all 36 diagnostic samples. All cases expressed CD179a in the end-induction B-lymphoblast population. Compared to gene-expression based predictions, we found a significant difference between expected versus observed flow-based CD179a positivity (two-sided Fisher’s exact test, P< 0.001). We found that CD179a expression was observed in cases having E2A-PBX3, KMT2A, BCR-ABL1 and other re-arrangements that typify mixed phenotype acute leukemias (MPALs). Conclusions: Our results show that CD179a is commonly expressed in B-ALL, regardless of stage, NCI risk features, or molecular aberrations. Because the productively assembled preBCR mediates autonomous survival signaling in pro- and pre-B cells, it may also contribute to the mechanistic basis of MRD in B-ALL. Immunotherapies directed against the CD179a component of the preBCR may spare the immune-compromise that occurs with pan B-cell ablation, and prevent the emergence of therapy-resistant disease in B-ALL/B-LLy.

New Immunophenotypic Profile of Waldenstrom’s Macroglobulinemia: Interest of CD80 and CD86 Staining

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 5277-5277
Author(s):  
Andrea Toma ◽  
Magali Le Garff-Tavernier ◽  
Martine Brissard ◽  
Patrick Bonnemye ◽  
Lucille Musset ◽  
...  
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
B Cell ◽  
Cell Population ◽  
Light Chain ◽  
Waldenström’S Macroglobulinemia ◽  
Waldenstrom's Macroglobulinemia ◽  
Waldenstrom’S Macroglobulinemia ◽  
Waldenström's Macroglobulinemia ◽  
Immunophenotypic Analysis

Abstract The immunophenotypic characterization is an essential tool in the diagnosis of hematological malignancies but the immunophenotypic features in Waldenstrom’s macroglobulinemia (WM) remain not clearly defined. We studied 96 cases of WM diagnosed by monoclonal IgM in the serum and morphological lymphoplasmacytic bone marrow infiltration, and we compared results to 33 cases of other chronic B-cell lymphoproliferative disorders (LPD), including marginal zone (MZL)(n=23), mantle cell (MCL)(n=8) and follicular (FL)(n=2) lymphomas. Patients with a Matutes score &gt;3 (chronic lymphocytic leukemia) and with pathognomonic immunophenotype (hairy cell leukemia) were excluded. Immunophenotypic analysis was performed by flow cytometry using six-colour staining (FACS Canto II, Becton Dickinson). In WM and LPD groups, a monoclonal B-cell population was identified in blood (31 and 28 patients, respectively), blood and bone marrow (28 and 4 patients) or bone marrow samples (23 and 1 patients). Overall, 61% of WM patients showed a monoclonal B-cell population in blood. Neoplastic cells of WM and LPD patients with blood and/or bone marrow involvement expressed a monoclonal immunoglobulin light chain kappa (in 70% and 73% of cases respectively) or lambda (30% and 27%). The intensity of expression of the light chain was heterogeneous in both groups (high, normal or low expression in 43%, 27% or 30% of WM, and in 52%, 33% or 15% of LPD, respectively). All pan-B antigens (CD20, CD19, CD79b) were positive for at least 97% of patients. Results obtained with other antigens in WM compared to LPD were: CD10 = 10% vs 7% of patients, CD23 = 33% vs 56%, CD5 = 14% vs 26%, FMC7 = 76% vs 89%, CD38 = 56% vs 41%, CD25 = 86% vs 84%, CD43 = 12% vs 16%, and CD11c = 10% vs 36%. The intensity of expression of these antigens was heterogeneous in both groups. Among the antigens only tested in the WM group, CD1c and CD27 were positive for 70% of patients, IgM and IgD for 95% of patients, and CD103 as well as CD117 were negative in all cases. No difference was found between blood and bone marrow for all previous antigens. Plasma cells (CD38/CD138 positive cells) were found at low levels (less than 2.5% of B-cells) for 46% of WM in blood and/or bone marrow samples. Among the 10 WM patients tested for ZAP-70 expression, 9 were negative and 1 showed a low intensity expression. These results confirm that the immunophenotypic analysis usually performed with standard antigens does not allow defining a typical profile of WM. In order to tentatively identify the WM among the B-cell malignancies, we studied the expression of molecules known to be involved in B-cell development or in costimulatory pathways of antigenic activation, namely CD69, CD83, CD80 and CD86. We first analyzed blood samples of 24 WM patients showing a peripheral monoclonal B-cell population. CD80 was positive (&gt; 20% of B-cells) in all cases and CD83, CD69 and CD86 were always negative. Among these WM patients, 13 were also studied for the bone marrow phenotype. No difference was found between blood and bone marrow phenotype in 11/13 WM cases. We then studied 11 LPD with blood tumoral involvement (MZL(n=7), MCL(n=2) and FL(n=2)). In these LPD, CD69 and CD83 were always negative and, in most cases (9/11 patients), CD80 and CD86 were also negative. Interestingly, CD80 was found positive in 2 patients with MZL, but the CD80 positivity was always associated to the CD86 positivity. Altogether, these data suggest that the inclusion of CD80 and CD86 in the panel of cytometric analysis allow to discriminate WM from other B-LPD with peripheral blood involvement.

Extensive in vivo self-renewal, long-term reconstitution capacity, and hematopoietic multipotency of Pax5-deficient precursor B-cell clones

Blood ◽  
2002 ◽  
Vol 99 (8) ◽  
pp. 2760-2766 ◽  
Author(s):  
Christoph Schaniel ◽  
Marie Gottar ◽  
Eddy Roosnek ◽  
Fritz Melchers ◽  
Antonius G. Rolink
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
B Cell ◽  
Telomerase Activity ◽  
Self Renewal ◽  
Cell Clones ◽  
Precursor B Cells

Abstract Self-renewal, pluripotency, and long-term reconstitution are defining characteristics of single hematopoietic stem cells.Pax5−/− precursor B cells apparently possess similar characteristics. Here, using serial transplantations, with in vitro recloning and growth of the bone marrow–homed donor cells occurring after all transplantations, we analyzed the extent of self-renewal and hematopoietic multipotency ofPax5−/− precursor B-cell clones. Moreover, telomere length and telomerase activity in these clones was analyzed at various time points. Thus far, 5 successive transplantations have been performed. Clones transplanted for the fifth time, which have proliferated for more than 150 cell divisions in vitro, still repopulate the bone marrow with precursor B cells and reconstitute these recipients with lymphoid and myeloid cells. During this extensive proliferation, Pax5−/− precursor B cells shorten their telomeres at 70 to 90 base pairs per division. Their telomerase activity remains at 3% of that of HEK293 cancer cells during all serial in vivo transplantations/in vitro expansions. Together, these data show thatPax5−/− precursor B-cell clones possess extensive in vivo self-renewal capacity, long-term reconstitution capacity, and hematopoietic multipotency, with their telomeres shortening at the normal rate.

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