Analysis of Immunoglobulin Gene Mutation Status in Lymphoplasmacytic and Splenic Marginal Zone Lymphomas with Bone Marrow Infiltration.

Abstract Splenic Marginal-Zone lymphomas (SMZL) and Lymphoplasmacytic lymphomas (LPL) correspond to entities that shared clinical, morphological and immunohistochemical features and are not always easy to discriminate. Immunoglobulin variable heavy chain gene (VH) usage and somatic mutation pattern were analyzed to clarify the relationship and the pathogenesis of these 2 entities. Bone marrow frozen samples from 27 patients, 16 LPL and 11 SMZL, were studied. B-cell clonality was first analysed using the standard PCR method with the Biomed 2 primers. Informative PCR products were then purified, sequenced and analyzed on IMGT and Ig-Blast websites. Overall, median rate of somatic mutations was 9% in LPL and 4% in SMZL (p=0.013). All cases of LPL were mutated. Mutation rate was always higher than 5%. A preferential usage of VH3 gene was observed in 13/16 cases (80%), with a predominance of VH3–23 and VH3–74, both representing 60% of cases. VH4 gene was used only in 2/16 cases (13%). Regarding JH usage, a predominance of JH4 gene rearrangement was observed in 11/16 cases (69%). JH5 and JH6 were used in 20% and 13% of cases respectively. In the group of SMZL, mutation rates and usage of VH and JH genes were more heterogeneous. Thirty six percent of SMZL were unmutated and 19% had a mutation rate lower than 5%. VH3 was used in 4/11 cases (36%), VH4 in 3/11 cases (27%) and VH1 in 4/11 cases (36%). JH4 and JH6 were used in 4/11 cases (36%) and the JH5 in 1/11 cases (9%). Therefore, the profile of somatic mutations and the VH and JH segment preferentially used are different in SMZL and LPL, suggesting a different process of antigen-driven selection and of transformation of the initial B-cell clone. These findings consolidate the existence of LPL as a genuine entity, different of SMZL despite their histopathological overlaps.

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A single pre-B cell can give rise to antigen-specific B cells that utilize distinct immunoglobulin gene rearrangements.

Journal of Experimental Medicine ◽

10.1084/jem.172.3.815 ◽

1990 ◽

Vol 172 (3) ◽

pp. 815-825 ◽

Cited By ~ 8

Author(s):

A J Caton

Keyword(s):

B Cells ◽

B Cell ◽

Cell Clone ◽

Gene Segment ◽

Chain Gene ◽

Gene Rearrangements ◽

Immunoglobulin Gene ◽

Immune Repertoire ◽

Influenza Virus Hemagglutinin ◽

Immunoglobulin Gene Rearrangements

A group of hybridomas that express antibodies with related specificities for the influenza virus hemagglutinin (HA), that represent B cells that were the clonal progeny of a single pre-B cell, and that utilized distinct L chain gene rearrangements have been characterized. The clonal relationship was established by the sharing of H chain gene rearrangements at both the productive and the nonproductive alleles. Among these hybridomas, one group had rearranged only one of its kappa alleles, having joined a V kappa 24 gene to the J kappa 2 gene segment. The other group utilized the same V kappa 24 gene segment in productive rearrangement to the J kappa 5 gene segment, and shared an aberrant rearrangements among members of the same B cell clone can normally occur, and can contribute to the generation and diversification of the immune repertoire that is available for the recognition of foreign antigens. Mechanisms by which the distinct rearrangements expressed by the hybridomas might have been generated are discussed.

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Acute undifferentiated leukemia: implications for cellular origin and clonality suggested by analysis of surface markers and immunoglobulin gene rearrangement

Blood ◽

10.1182/blood.v68.3.658.658 ◽

1986 ◽

Vol 68 (3) ◽

pp. 658-662 ◽

Cited By ~ 47

Author(s):

A Raghavachar ◽

CR Bartram ◽

A Ganser ◽

G Heil ◽

E Kleihauer ◽

...

Keyword(s):

T Cell ◽

B Cell ◽

Cell Clone ◽

Cell Lineage ◽

Surface Marker ◽

Leukemic Cells ◽

Chain Gene ◽

Surface Markers ◽

Immunoglobulin Gene ◽

Cellular Origin

Abstract Although B cell leukemias and, recently, T cell leukemias can be identified both by surface marker and molecular analysis, there remains a population of acute undifferentiated leukemias (AUL) that cannot be allocated definitively to a single cell lineage. AUL was diagnosed in nine patients according to stringent criteria. We combined both immunologic and molecular approaches to analyze further the ambiguous origin of AUL cells. Southern blot analysis revealed rearranged Ig heavy-chain genes in seven patients and indicated a biclonal or oligoclonal leukemic cell population in three of them, including one case of AUL with translocation (4;11). Analysis of cell surface markers showed expression of at least one early B cell-associated antigen (BA- 1, BA-2, B4, UL-38) in six of these seven patients, with coexpression of a myeloid antigen (VIM-2) in three patients. Leukemic cells of two other patients neither exhibited Ig chain gene rearrangements nor expressed B cell-associated antigens. T cell receptor beta-chain genes showed germline configuration in all nine cases. Our results demonstrate heterogeneity among AUL patients based on molecular and surface marker analyses and suggest that most AUL blast cells are derived from a precursor cell that shares phenotypic and genotypic characteristics of early B cells with certain surface antigens of myeloid cells, in some cases of AUL more than one abnormal cell clone or subclone may exist, and the cellular origin, at least of some AULs exhibiting t(4;11), may be truly B cell lineage committed.

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Acute undifferentiated leukemia: implications for cellular origin and clonality suggested by analysis of surface markers and immunoglobulin gene rearrangement

Blood ◽

10.1182/blood.v68.3.658.bloodjournal683658 ◽

1986 ◽

Vol 68 (3) ◽

pp. 658-662

Author(s):

A Raghavachar ◽

CR Bartram ◽

A Ganser ◽

G Heil ◽

E Kleihauer ◽

...

Keyword(s):

T Cell ◽

B Cell ◽

Cell Clone ◽

Cell Lineage ◽

Surface Marker ◽

Leukemic Cells ◽

Chain Gene ◽

Surface Markers ◽

Immunoglobulin Gene ◽

Cellular Origin

Although B cell leukemias and, recently, T cell leukemias can be identified both by surface marker and molecular analysis, there remains a population of acute undifferentiated leukemias (AUL) that cannot be allocated definitively to a single cell lineage. AUL was diagnosed in nine patients according to stringent criteria. We combined both immunologic and molecular approaches to analyze further the ambiguous origin of AUL cells. Southern blot analysis revealed rearranged Ig heavy-chain genes in seven patients and indicated a biclonal or oligoclonal leukemic cell population in three of them, including one case of AUL with translocation (4;11). Analysis of cell surface markers showed expression of at least one early B cell-associated antigen (BA- 1, BA-2, B4, UL-38) in six of these seven patients, with coexpression of a myeloid antigen (VIM-2) in three patients. Leukemic cells of two other patients neither exhibited Ig chain gene rearrangements nor expressed B cell-associated antigens. T cell receptor beta-chain genes showed germline configuration in all nine cases. Our results demonstrate heterogeneity among AUL patients based on molecular and surface marker analyses and suggest that most AUL blast cells are derived from a precursor cell that shares phenotypic and genotypic characteristics of early B cells with certain surface antigens of myeloid cells, in some cases of AUL more than one abnormal cell clone or subclone may exist, and the cellular origin, at least of some AULs exhibiting t(4;11), may be truly B cell lineage committed.

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Aggressive B-cell lymphomas in patients with myelofibrosis receiving JAK1/2 inhibitor therapy

Blood ◽

10.1182/blood-2017-10-810739 ◽

2018 ◽

Vol 132 (7) ◽

pp. 694-706 ◽

Cited By ~ 59

Author(s):

Edit Porpaczy ◽

Sabrina Tripolt ◽

Andrea Hoelbl-Kovacic ◽

Bettina Gisslinger ◽

Zsuzsanna Bago-Horvath ◽

...

Keyword(s):

Bone Marrow ◽

B Cell ◽

Cell Clone ◽

Myeloproliferative Neoplasms ◽

Fold Increase ◽

Inhibitor Therapy ◽

Janus Kinase ◽

Immunoglobulin Gene ◽

B Cell Lymphomas ◽

Increased Risk

Inhibition of Janus-kinase 1/2 (JAK1/2) is a mainstay to treat myeloproliferative neoplasms (MPN). Sporadic observations reported the co-incidence of B-cell non-Hodgkin lymphomas during treatment of MPN with JAK1/2 inhibitors. We assessed 626 patients with MPN, including 69 with myelofibrosis receiving JAK1/2 inhibitors for lymphoma development. B-cell lymphomas evolved in 4 (5.8%) of 69 patients receiving JAK1/2 inhibition compared with 2 (0.36%) of 557 with conventional treatment (16-fold increased risk). A similar 15-fold increase was observed in an independent cohort of 929 patients with MPN. Considering primary myelofibrosis only (N = 216), 3 lymphomas were observed in 31 inhibitor-treated patients (9.7%) vs 1 (0.54%) of 185 control patients. Lymphomas were of aggressive B-cell type, extranodal, or leukemic with high MYC expression in the absence of JAK2 V617F or other MPN-associated mutations. Median time from initiation of inhibitor therapy to lymphoma diagnosis was 25 months. Clonal immunoglobulin gene rearrangements were already detected in the bone marrow during myelofibrosis in 16.3% of patients. Lymphomas occurring during JAK1/2 inhibitor treatment were preceded by a preexisting B-cell clone in all 3 patients tested. Sequencing verified clonal identity in 2 patients. The effects of JAK1/2 inhibition were mirrored in Stat1−/− mice: 16 of 24 mice developed a spontaneous myeloid hyperplasia with the concomitant presence of aberrant B cells. Transplantations of bone marrow from diseased mice unmasked the outgrowth of a malignant B-cell clone evolving into aggressive B-cell leukemia-lymphoma. We conclude that JAK/STAT1 pathway inhibition in myelofibrosis is associated with an elevated frequency of aggressive B-cell lymphomas. Detection of a preexisting B-cell clone may identify individuals at risk.

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IRF4 Has a Unique Role in Early B Cell Development and Acts Prior to CD21 Expression to Control Marginal Zone B Cell Numbers

Frontiers in Immunology ◽

10.3389/fimmu.2021.779085 ◽

2021 ◽

Vol 12 ◽

Author(s):

Kristina Ottens ◽

Anne B. Satterthwaite

Keyword(s):

Bone Marrow ◽

B Cells ◽

B Cell ◽

Marginal Zone ◽

Cell Development ◽

B Cell Development ◽

Chain Gene ◽

Unique Role ◽

Cell Numbers ◽

Transitional B Cells

Strict control of B lymphocyte development is required for the ability to mount humoral immune responses to diverse foreign antigens while remaining self-tolerant. In the bone marrow, B lineage cells transit through several developmental stages in which they assemble a functional B cell receptor in a stepwise manner. The immunoglobulin heavy chain gene is rearranged at the pro-B stage. At the large pre-B stage, cells with a functional heavy chain expand in response to signals from IL-7 and the pre-BCR. Cells then cease proliferation at the small pre-B stage and rearrange the immunoglobulin light chain gene. The fully formed BCR is subsequently expressed on the surface of immature B cells and autoreactive cells are culled by central tolerance mechanisms. Once in the periphery, transitional B cells develop into mature B cell subsets such as marginal zone and follicular B cells. These developmental processes are controlled by transcription factor networks, central to which are IRF4 and IRF8. These were thought to act redundantly during B cell development in the bone marrow, with their functions diverging in the periphery where IRF4 limits the number of marginal zone B cells and is required for germinal center responses and plasma cell differentiation. Because of IRF4’s unique role in mature B cells, we hypothesized that it may also have functions earlier in B cell development that cannot be compensated for by IRF8. Indeed, we find that IRF4 has a unique role in upregulating the pre-B cell marker CD25, limiting IL-7 responsiveness, and promoting migration to CXCR4 such that IRF4-deficient mice have a partial block at the pre-B cell stage. We also find that IRF4 acts in early transitional B cells to restrict marginal zone B cell development, as deletion of IRF4 in mature B cells with CD21-cre impairs plasma cell differentiation but has no effect on marginal zone B cell numbers. These studies highlight IRF4 as the dominant IRF family member in early B lymphopoiesis.

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Clustering of extensive somatic mutations in the variable region of an immunoglobulin heavy chain gene from a human B cell lymphoma

Cell ◽

10.1016/0092-8674(86)90488-5 ◽

1986 ◽

Vol 44 (1) ◽

pp. 97-106 ◽

Cited By ~ 173

Author(s):

Michael L. Cleary ◽

Timothy C. Meeker ◽

Shoshana Levy ◽

Elizabeth Lee ◽

Martha Trela ◽

...

Keyword(s):

B Cell ◽

Heavy Chain ◽

Somatic Mutations ◽

Cell Lymphoma ◽

Variable Region ◽

B Cell Lymphoma ◽

Chain Gene ◽

Heavy Chain Gene ◽

Immunoglobulin Heavy Chain Gene ◽

Human B Cell

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Disease-biased and shared characteristics of the immunoglobulin gene repertoires in marginal zone B cell lymphoproliferations

The Journal of Pathology ◽

10.1002/path.5209 ◽

2019 ◽

Vol 247 (4) ◽

pp. 416-421 ◽

Cited By ~ 7

Author(s):

Aliki Xochelli ◽

Vasilis Bikos ◽

Eleftheria Polychronidou ◽

Chrysi Galigalidou ◽

Andreas Agathangelidis ◽

...

Keyword(s):

B Cell ◽

Marginal Zone ◽

Immunoglobulin Gene

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Emergence of a B-cell lymphoblastic lymphoma in a patient with B-cell chronic lymphocytic leukemia: evidence for the single-cell origin of the two tumors

Blood ◽

10.1182/blood.v78.3.797.bloodjournal783797 ◽

1991 ◽

Vol 78 (3) ◽

pp. 797-804

Author(s):

V Pistoia ◽

S Roncella ◽

PF Di Celle ◽

M Sessarego ◽

G Cutrona ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

B Cell ◽

Cell Lines ◽

Peripheral Blood ◽

Chromosomal Abnormalities ◽

Cell Clone ◽

Lymphoblastic Lymphoma ◽

Lymphocytic Leukemia ◽

Terminal Deoxynucleotidyl Transferase ◽

Chain Gene

A patient is described who presented with a chronic lymphocytic leukemia (CLL) and later developed a lymphoblastic lymphoma. The cells from the CLL were typical mature B lymphocytes as could be assessed by morphologic, cytochemical, and surface marker analyses. The cells from the lymphoblastic lymphoma were immature B cells that expressed CD10, CD20, and HLA-DR markers, but not surface Ig or cytoplasmic mu chains, and were negative for terminal deoxynucleotidyl transferase (TdT). The cells of two continuous cell lines, obtained from the bone marrow and the peripheral blood of the patient, had the same phenotype as the lymphoblastic lymphoma cells, did not contain the Epstein-Barr virus genome, and displayed malignant features in vitro, including the capacity to form colonies in agar. The two cell lines also shared identical chromosomal abnormalities, a finding which suggests that they derived from the same malignant cell already present in vivo. Such chromosomal abnormalities were not seen in the karyotype of the peripheral blood cells at the onset of the disease. Analysis of the Ig heavy chain genes using a DJ-specific probe showed the very same monoclonal rearrangement in the cells from the B-CLL, the lymphoblastic lymphoma and the two cell lines, thus demonstrating their common clonal origin. By contrast, a monoclonal rearrangement of the lambda chain gene locus was found in the B-CLL cells only, a finding consistent with their exclusive capacity to express surface IgM lambda. This patient represents a rare case in whom a chronic lymphoproliferative disorder with mature malignant cells transforms into a lymphoblastic lymphoma characterized by cells frozen at a very early maturational stage. The possible mechanisms leading to such transformation within the same cell clone are discussed.

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