Somatic mutation analysis of IgH variable regions reveals that tumor cells of most parafollicular (monocytoid) B-cell lymphoma, splenic marginal zone B-cell lymphoma, and some hairy cell leukemia are composed of memory B lymphocytes

doi:10.1016/S0046-8177(99)90010-2

Human Pathology

Volume 30, Issue 3, March 1999, Pages 306-312

https://doi.org/10.1016/S0046-8177(99)90010-2 Get rights and content

Abstract

The cell of origin of parafollicular (monocytoid) B cell lymphoma (PBCL), splenic marginal zone lymphoma (SMZL), and hairy cell leukemia (HCL) is controversial. To better understand the relationship between these low-grade B-cell neoplasms, we analyzed the nucleotide sequences of the rearranged immunoglobulin heavy (IgH) chain variable (V) region of the clonal population of cells in five cases of PBCL, four cases of SMZL, and seven cases of HCL to determine whether these neoplasms could be differentiated by the degree of somatic mutation in the IgH V gene or by the IgH V gene family usage. DNA was extracted from diagnostic material and clonality confirmed by PCR. The DNA was reamplified using V heavy chain family specific primers, and the amplicons were sequenced. Sequences were compared with germline IgH V gene sequences, and base changes were determined to be silent or to represent amino acid replacements by using three different methods. Four of five (80%) cases of PBCL, three of four (75%) cases of SMZL, and three of seven (43%) cases of HCL showed evidence of antigen selection, suggesting that these neoplasms involved clonal expansions of postgerminal center memory lymphocytes. Only SMZL showed a preferential usage of V_H1 family genes.

References (37)

AK Stewart et al.
Immunoglobulin V regions and the B cell
Blood
(1994)
SD Wagner et al.
Similar patterns of V_κ gene usage but different degrees of somatic mutation in hairy cell leukemia, prolymphocytic leukemia, Waldenstrom's macroglobulinemia, and myeloma
Blood
(1994)
MHC Bakkus et al.
Evidence that multiple myeloma Ig heavy chain VDJ genes contain somatic mutations but show no intraclonal variation
Blood
(1992)
R Kuppers et al.
B cells of chronic lymphatic leukemia express V genes in unmutated form
Leukemia Res
(1991)
NL Harris et al.
A revised European-American classification of lymphoid neoplasms: A proposal from the International Lymphoma Study Group
Blood
(1994)
ST Traweek et al.
Monocytoid B-cell lymphoma: Its evolution and relationship to other low-grade B-cell neoplasms
Blood
(1989)
F Fend et al.
Monocytoid B-cell lymphoma: Its relationship to and possible cellular origin from marginal zone cells
Hum Pathol
(1993)
GH Segal et al.
Optimal primer selection for clonality assessment by polymerase chain reaction analysis: I. Low grade B-cell lymphoproliferative disorders of nonfollicular center cell type
Hum Pathol
(1994)
MJ Campbell et al.
Use of family specific leader region primers for PCR amplification of the human heavy chain variable region gene repertoire
Mol Immunol
(1992)
SF Altschul et al.
Basic local alignment search tool
J Mol Biol
(1990)

F Davi et al.

High frequency of somatic mutations in the V_H genes expressed in prolymphocytic leukemia

Blood

(1996)

B Chang et al.

The CDR1 sequences of a major proportion of human germline IgV_H genes are inherently susceptible to amino acid replacement

Immunol Today

(1994)

N Maizels

Somatic hypermutation: How many mechanisms diversify V region sequences?

Cell

(1995)

Y Qin et al.

Somatic hypermutation in low-grade mucosa-associated lymphoid tissue-type B-cell lymphoma

Blood

(1995)

D Zhu et al.

Splenic lymphoma with villous lymphocytes involves B cells with extensively mutated Ig heavy chain variable region genes

Blood

(1995)

RJ Lukes et al.

Tumors of the hematopoietic system

RA Warnke et al.

Tumors of the lymph nodes and spleen

RA Insel et al.

Somatic mutation of human immunoglobulin V genes: Bias, rate and regulation

Ann NY Acad Sci

(1995)

Cited by (56)

Small cell lymphocytic variant of marginal zone lymphoma: A distinct form of marginal zone lymphoma derived from naïve B cells as a cutaneous counterpart to the naïve marginal zone lymphoma of splenic origin
2018, Annals of Diagnostic Pathology
Citation Excerpt :
In general, female patients were younger (median age = 58 years) than males (median age = 75 years). Follow up was obtained on 9 of the 11 cases [1,3-5,7-11]. At the time of the initial presentation, all patients were feeling well without constitutional symptoms and presented with limited disease in the context of a single lesion in 6 cases, 2 lesions in one anatomic region on the face in 1 case and multiple lesions of discontiguous regions in two cases.
Treatment of splenic marginal zone lymphoma
2017, Best Practice and Research: Clinical Haematology
Splenic marginal zone lymphoma (SMZL) is a distinct lymphoma entity characterized by an indolent clinical course and prolonged survival. Treatment is not standardized, since there are no prospective randomized trials in large series of SMZL patients. Splenectomy and rituximab represent the most effective treatment strategies used so far. The addition of chemotherapy to rituximab has not further improved the outcome, although this issue requires further investigation. Rituximab monotherapy has been associated with high response rates (∼90%), with approximately half of these responses being complete, even at the molecular level. More importantly, many of these responses are long-lasting, with a reported 7-year progression-free survival (PFS) at the rate of 69%. Maintenance rituximab treatment has been associated with further improvement of the quality of response as well as longer response duration in studies derived from one group of investigators. Based on its high efficacy and the good safety profile, rituximab represent one of the best treatment options for SMZL patients. Moreover, rituximab retains its efficacy in the relapse setting in most cases. Splenectomy is a meaningful alternative to rituximab in patients with bulky splenomegaly and cytopenias, without extensive bone marrow infiltration, who are fit for surgery. However splenectomy cannot completely eradicate the disease and it is also associated with greater morbidity or even mortality compared to rituximab. The choice of one of these two treatment approaches (rituximab or splenectomy) should mainly be based on patient's characteristics and on the disease burden. Novel agents are currently testing in low grade lymphomas including a small number of SMZL patients with promising results.
Evidence of canonical somatic hypermutation in hairy cell leukemia
2011, Blood
Citation Excerpt :
The presence of canonical SHM in both normal and malignant B cells has been considered as strong evidence of an antigen-driven mutational process.5,6 We and others have previously analyzed SHM in HCL and in other situations in terms of related factors to detect an antigen-driven process, including total R/S ratio, CDR/FR ratio of R-mutations, and p-multinomial and p-binomial statistics.11,14,18,31-37 Our previous study of these statistics in 23 HCL patients identified only 2 patients with strong evidence (P = .001-.003) of antigen-driven SHM.11
To compare hairy cell leukemia (HCL) with chronic lymphocytic leukemia (CLL) and normal B cells with respect to their B-cell receptors, somatic hypermutation (SHM) features in HCL were examined in a series of 130 immunoglobulin gene heavy chain rearrangements, including 102 from 100 classic (HCLc) and 28 from 26 variant (HCLv) patients. The frequency of unmutated rearrangements in HCLc was much lower than that in HCLv (17% vs 54%, P < .001) or historically in CLL (17% vs 46%, P < .001), but HCLv and CLL were similar (P = .45). As previously reported for CLL, evidence of canonical SHM was observed in HCLc rearrangements, including: (1) a higher ratio of replacement to silent mutations in the complementarity determining regions than in the framework regions (2.83 vs 1.41, P < .001), (2) higher transition to transversion ratio than would be expected if mutations were random (1.49 vs 0.5, P < .001), and (3) higher than expected concentration of mutations within RGYW hot spots (13.92% vs 3.33%, P < .001). HCLv met these 3 criteria of canonical SHM to a lesser extent. These data suggest that, whereas HCLc cells may recognize antigen-like CLL and normal B cells before malignant transformation, HCLv cells from some patients may originate differently, possibly without undergoing antigen recognition.
VH4-34<sup>+</sup> hairy cell leukemia, a new variant with poor prognosis despite standard therapy
2009, Blood
Hairy cell leukemia variant (HCLv) presents with high disease burden, lack of typical antigens like CD25, and poor response to standard treatments like cladribine. Occasionally, patients with classic HCL respond poorly. Clinical and molecular features of HCL and HCLv has not been compared. Rearrangements expressing immunoglobulin VH chain were sequenced, including 22 from 20 patients with HCLv and 63 from 62 patients with classic HCL. Most patients were seeking relapsed/refractory trials, representing a poor-prognosis population. VH4-34, a gene commonly used in autoimmune disorders, was observed in 8 (40%) HCLv and 6 (10%) classic (P = .004) HCL patients. Compared with 71 VH4-34⁻ rearrangements, 14 VH4-34⁺ rearrangements were more frequently (P < .001) unmutated, defined as greater than 98% homologous to germline sequence. VH4-34⁺ patients had greater white blood cell counts at diagnosis (P = .002), lower response rate (P < .001) and progression-free survival (P = .007) after initial cladribine, and shorter overall survival from diagnosis (P < .001). Response and survival were more closely related to VH4-34 status than to whether or not patients had HCLv. VH4-34⁺ HCL is an important disorder that only partly overlaps with the previously described HCLv. Response to initial single-agent cladribine therapy is suboptimal; these patients should be considered for alternative approaches, including antibody-related therapy.
Splenic red pulp lymphoma with numerous basophilic villous lymphocytes: A distinct clinicopathologic and molecular entity?
2008, Blood
Citation Excerpt :
Although the present series does not resemble HCL, some overlaps exist. Indeed, common features include the male predominance, the massive congested red pulp pattern, the rarity of clonal chromosomal abnormalities, and the IgH mutational pattern (most cases mutated, low degree of intraclonal heterogeneity, and similar overrepresentation of VH3-23 and VH4-3426–30). In addition, immunophenotypic similarities include the expression of classical HCL markers (ie, CD11c, CD103, and CD123) but with a lower staining intensity than in HCL (Table 2) and a possible coexpression of preswitched (IgM/IgD) and postswitched (IgG) isotypes by the same cells (Table 2).
The presence of circulating villous lymphocytes (VLs) in lymphoma patients usually points to splenic marginal zone B-cell lymphoma (SMZL), even if the VLs can be found occasionally in other small B-cell lymphomas. However, those cells are variably described, and detailed cytologic characterization is often lacking. We identified lymphoma cases with numerous basophilic VLs among the large group of splenic lymphoma with VLs, and for further delineation, 37 cases with this particular cytology were analyzed. Patients, predominantly older men, presented with moderate lymphocytosis and splenomegaly without pancytopenia. The monoclonal B cells expressed IgM + D, IgM + G, IgM or IgG, as well as CD76 and CD11c, frequently CD103, and rarely CD123. Spleen sections were peculiar, with atrophic white pulp and a monomorphic diffuse lymphoma infiltration in a congested red pulp. Bone marrow infiltration was interstitial and intrasinusoidal without extensive fibrosis. Cytogenetic analysis showed a frequent absence of clonal aberrations (68%). Most cases (79%) were IgH mutated, with an overrepresentation of V_H3 and V_H4 gene families. These results, as well as the clinical evolution, show that those lymphoma cases represent a homogeneous group distinct from SMZL and reminiscent of hairy cell leukemia variant, perhaps corresponding to a separate lymphoma entity.
Molecular characterization of complete and incomplete immunoglobulin heavy chain gene rearrangements in hairy cell leukemia
2007, Clinical Lymphoma and Myeloma
We analyzed patients with hairy cell leukemia (HCL) to achieve a better understanding of the differentiation stage reached by HCL cells and to define the key role of the diversification of cell surface makers, especially CD25 expression.
We analyzed 38 previously untreated patients with HCL to characterize their complete (VDJ_H) and incomplete (DJ_H) immunoglobulin (Ig) heavy chain (IgH) rearrangements, including somatic hypermutation pattern and gene segment use.
A correlation between immunophenotypic profile and molecular data was seen. All 38 cases showed monoclonal amplifications: VDJ_H in 97%, DJ_H in 42%, and both in 39%. Segments from the D_H3 family were used more in complete compared with incomplete rearrangements (45% vs. 12%; P < .005). Furthermore, comparison between molecular and immunophenotypic characteristics disclosed differences in the expression of CD25 antigen; CD25^– cases, a phenotype associated with HCL variant, showed complete homology to the germline in 3 of 5 cases (60%), whereas this characteristic was never observed in CD25⁺ cases (P < .005). Moreover, V_H4-34, V_H1-08, and J_H3 segments appeared in 2, 1, and 2 CD25^– cases, respectively, whereas they were absent in all CD25⁺ cases.
These results support that HCL is a heterogeneous entity including subgroups with different molecular characteristics, which reinforces the need for additional studies with a larger number of patients to clarify the real role of gene rearrangements in HCL.

View all citing articles on Scopus

View full text

Original contributionSomatic mutation analysis of IgH variable regions reveals that tumor cells of most parafollicular (monocytoid) B-cell lymphoma, splenic marginal zone B-cell lymphoma, and some hairy cell leukemia are composed of memory B lymphocytes

Abstract

Blood

Blood

Blood

Leukemia Res

Blood

Blood

Hum Pathol

Hum Pathol

Mol Immunol

J Mol Biol

Blood

Immunol Today

Cell

Blood

Blood

Tumors of the hematopoietic system

Tumors of the lymph nodes and spleen

Somatic mutation of human immunoglobulin V genes: Bias, rate and regulation

Ann NY Acad Sci

Original contribution
Somatic mutation analysis of IgH variable regions reveals that tumor cells of most parafollicular (monocytoid) B-cell lymphoma, splenic marginal zone B-cell lymphoma, and some hairy cell leukemia are composed of memory B lymphocytes