B-cell gene rearrangement in benign and malignant lymphoid proliferations of mucosa-associated lymphoid tissue and lymph nodes

Torlakovic, Emina; Cherwitz, David L.; Jessurun, José; Scholes, John V.; McGlennen, Ronald C.

doi:10.1016/s0046-8177(97)90101-5

Cited by 62 publications

(53 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Nevertheless, some studies have demonstrated instances of false positivity, particularly when small or microdissected specimens are used, 18 and in reactive conditions involving extranodal sites such as the stomach and salivary gland. 19,20 The results of our study clearly show a relationship between a paucity of B cell targets and a tendency for generating pseudomonoclonal or pseudo-oligoclonal bands by IgH PCR. Using DNA obtained from reactive lymph nodes and tonsils, we found that polyclonal results were reliably obtained when the starting quantity of template DNA was 50 ng or more.…”

Section: Discussionmentioning

confidence: 54%

“…In this regard, others have shown that non-reproducible pseudomonoclonal bands can also be generated in DNA samples obtained from specimens containing abundant lymphocytic aggregates. 19,24 We recommend that for reliable assessment of IgH PCR-based clonality studies using DNA obtained from fixed, paraffin-embedded tissue extracted samples, no less than 50 ng of DNA should be used as the starting quantity of template DNA, of which B cells should account for at least 5 ng (10% of the total cells). We also recommend that multiple (at least two) aliquots of a DNA sample be subjected to IgH PCR analysis, and that specimens that produce pseudomonoclonal bands be categorized as indeterminate.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

PCR Analysis of the Immunoglobulin Heavy Chain Gene in Polyclonal Processes Can Yield Pseudoclonal Bands as an Artifact of Low B Cell Number

Elenitoba-Johnson¹,

Bohling²,

Mitchell³

et al. 2000

The Journal of Molecular Diagnostics

View full text Add to dashboard Cite

Polymerase chain reaction (PCR)-based analysis for detecting immunoglobulin heavy chain gene (IgH)rearrangements in lymphoproliferative disorders is well established. The presence of one or two discrete bands is interpreted as a monoclonal proliferation, whereas a smear pattern represents a polyclonal population. Prompted by our observation of discrete bands in histologically reactive processes with a relative paucity of B cells, we sought to determine whether low numbers of B cells in biopsy specimens could artifactually produce pseudomonoclonal bands. We performed IgH PCR analysis on serially diluted DNA samples from 5 B cell non-Hodgkin's lymphomas (B-NHLs), 5 reactive lymph nodes, 5 reactive tonsils and 10 microdissected germinal centers from a lymph node with follicular hyperplasia. We also assessed multiple aliquots of DNA samples from small biopsy specimens of reactive lymphocytic processes from the stomach (5 cases). PCR products were evaluated using high resolution agarose or polyacrylamide gels, and DNA sequencing was performed on IgH PCR products from two reactive germinal centers, which yielded monoclonal bands of identical size. The utility of molecular techniques such as Southern blot hybridization (SBH) analysis and polymerase chain reaction (PCR) in the assessment of clonality in lymphoproliferative disorders is well established. Although SBH is a fairly sensitive and specific method, its utility as a routine diagnostic tool is hampered by its requirement for high quality DNA, its labor-intensive nature, and its consequent long turnaround time. On the other hand, the rapid and less labor-intensive PCR continues to gain popularity as a technique for the assessment of clonality in lymphoproliferative disorders. 1,2 The results obtained with antigen receptor gene PCR correlate well with the results of SBH. [3][4][5] PCR-based assays have additional advantages over SBH analysis. PCR requires smaller quantities of DNA, and high molecular weight DNA is not necessary. Therefore, PCR has been applied to small biopsies and fixed paraffin-embedded tissues, which are generally unsuitable for SBH analysis. 6 PCR is extremely sensitive and can detect 1 positive cell in a background of 10 5 negative cells. Immunoglobulin heavy chain (IgH) PCR is capable of detecting 1 monoclonal B cell in a background of 10 2 to 10 3 polyclonal B cells. 7 The extreme sensitivity of PCR also constitutes a potential source for pitfalls in the interpretation of PCR-based antigen receptor gene rearrangement studies. Clearly, contamination is a frequent concern. Additionally, we have observed discrete bands in samples obtained from small biopsy specimens in which histological and immunophenotypic evaluation revealed a reactive process. We believe that the discrete bands generated in this situation are related to the paucity of B

show abstract

Section: Discussionmentioning

confidence: 54%

Section: Discussionmentioning

confidence: 99%

PCR Analysis of the Immunoglobulin Heavy Chain Gene in Polyclonal Processes Can Yield Pseudoclonal Bands as an Artifact of Low B Cell Number

Elenitoba-Johnson¹,

Bohling²,

Mitchell³

et al. 2000

The Journal of Molecular Diagnostics

View full text Add to dashboard Cite

show abstract

“…This may explain why most previous studies found evidence for the presence of only a single tumor cell population in gastric lymphomas (Aiello et al, 1999;Nakamura et al, 1998;Peng et al, 1997). Also, interpretation of PCR products of differing sizes as being polyclonal (Torlakovic et al, 1997) may explain why oligoclonality has not been detected more frequently in gastric lymphomas.…”

Section: Discussionmentioning

confidence: 95%

Biclonality of Gastric Lymphomas

Cabras

Candidus

Fend

et al. 2001

Laboratory Investigation

View full text Add to dashboard Cite

SUMMARY:The pathogenesis and clonal evolution of gastric diffuse large B-cell lymphoma (DLBCL) and its relationship to extranodal marginal zone B-cell lymphoma (MZBL), mucosa-associated lymphoid tissue (MALT) type, are still controversial. The aim of this study was to establish the clonality of morphologically distinct areas of gastric lymphomas as well as their genetic relationship to each other. Six gastric lymphomas, consisting of two MZBL, MALT type, two DLBCL, and two "composite" lymphomas were subjected to laser capture microdissection and subsequent PCR-based amplification of the immunoglobulin heavy chain gene. One DLBCL showed a biclonal pattern of rearranged immunoglobulin heavy chain (IgH) genes of two different areas without evidence of a common origin. Two composite DLBCL with areas of extranodal MZBL, MALT type, were also biclonal and displayed different IgH gene rearrangements in the small-cell and in the large-cell components, respectively. Sequencing of the CDR3 region revealed unique VH-N-D and D-N-JH junctions, thus corroborating the presence of two genuinely distinct tumor clones in each of these three cases. In contrast, the remaining three gastric lymphomas (one DLBCL and two MZBL, MALT type) showed IgH gene rearrangements in which CDR3 regions were identical in the different tumor areas. Our results suggest that gastric DLBCL may be composed of more than one tumor cell clone. Further, DLBCL may not necessarily evolve by transformation of a low-grade lymphoma, but may also originate de novo. An ongoing emergence of new tumor clones may considerably hamper molecular diagnosis and follow-up of gastric DLBCL. (Lab Invest 2001, 81:961-967).T he two main groups of gastric lymphoma are extranodal marginal zone B-cell lymphoma (MZBL), mucosa-associated lymphoid tissue (MALT) type, and diffuse large B-cell lymphoma (DLBCL) . MZBL, MALT type, is composed of a monotonous, diffuse infiltrate of small lymphoid cells. DLBCL is characterized by a diffuse infiltrate of large malignant lymphoid blasts resembling centroblasts, plasmablasts, and immunoblasts. Occasionally, DLBCL shows both small-and large-cell components. These tumors have been called "composite" lymphomas, their histologic spectrum ranging from cases with well separated small-and large-cell components, to tumors in which these components are intermingled (De Jong et al, 1997;De Wolf-Peeters and Achten, 1999).The histogenesis of gastric DLBC lymphoma, as well as its possible clonal relationship to concomitant or preceding low-grade components, has been strongly debated recently and is still not fully clarified. The clonality of B-cell non-Hodgkin's lymphomas can be assessed by the PCR analysis of rearranged immunoglobulin heavy chain (IgH) genes that vary in size in different neoplastic cell populations (Trainor et al, 1991). Several studies comparing the patterns of clonal IgH gene rearrangement between large-cell and small-cell areas in the same gastric B-cell lymphoma have demonstrated a common clonal origin of both components (Chan et al...

show abstract

“…Torlakovic et al (23) reported an indefinite pattern of oligoclonal bands in MALT lymphoma and explained these findings to be caused by a selective proliferation of relatively small number of B-cell clones responsive to H. pylori. With respect to histology, 75% of the present lesions with largecell type showed monoclonal or biclonal pattern proliferation, indicating the advanced stage of MALT lymphoma.…”

Section: Discussionmentioning

confidence: 99%

Clonal Evolution of Gastric Lymphoma of Mucosa-Associated Lymphoid Tissue Type

et al. 2001

View full text Add to dashboard Cite

Development of multiple lesions is frequent in gastric lymphoma of mucosa-associated lymphoid tissue (MALT) type. Presence of clonal components in multiple lesions was examined on the resected samples from 18 cases by using PCR-based method for immunoglobulin heavy-chain gene rearrangement. There were two or more lesions in 10 cases, and 2 to 12 samples were obtained from each lesion. The remaining eight cases had a single large lesion, from which two to six samples were collected from separate areas from each other. A total of 86 samples were analyzed. Histologic findings in each sample were categorized as follows: proliferation of exclusively centrocyte-like cells (CCL), large cells, and combined CCL and large cells. Monoclonal or biclonal pattern (single or two bands) was observed in 42 samples, oligoclonal pattern (three or more bands) in 30, polyclonal (smear) in 11, and no products in 3. Large-cell-type lesions showed fewer bands than those with other histologic types, and 75% of cases with large-cell type had mono-or biclonal proliferation. Common clones were found among lesions in about 60% of cases. Especially in 4 cases including 2 cases with large-cell type, every lesion in the same case contained the common clones. These findings suggested that gastric MALT lymphoma started as multi-or oligoclonal proliferation of cells, in which separate lesions composed of different clones from each other. As disease advances, dominant clones appear in some lesion and disseminate to other lesions via homing properties of the proliferating B lymphocytes.

show abstract

B-cell gene rearrangement in benign and malignant lymphoid proliferations of mucosa-associated lymphoid tissue and lymph nodes

Cited by 62 publications

References 44 publications

PCR Analysis of the Immunoglobulin Heavy Chain Gene in Polyclonal Processes Can Yield Pseudoclonal Bands as an Artifact of Low B Cell Number

PCR Analysis of the Immunoglobulin Heavy Chain Gene in Polyclonal Processes Can Yield Pseudoclonal Bands as an Artifact of Low B Cell Number

Biclonality of Gastric Lymphomas

Clonal Evolution of Gastric Lymphoma of Mucosa-Associated Lymphoid Tissue Type

Contact Info

Product

Resources

About