Determination of B-Cell Clonality in Paraffin-Embedded Lymph Nodes Using the Polymerase Chain Reaction

Reed, Thomas J; Reid, Ann; Wallberg, Karen; O??Leary, Timothy J.; Frizzerà, Glauco

doi:10.1097/00019606-199303000-00007

Cited by 83 publications

(30 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…[45][46][47][48] DNA was extracted from formalinfixed, paraffin-embedded tissues in all cases and purified according to previously published methods. 46,48 Consensus primers were used for detection of IgH chain (FR3A V region and CFW1 J region), TCRb chain (VJ1, VJ2, D1J2, and D2J2), and TCRg chain (V2, V3, V4, V8, V9, JGT3, JGT4, and JGT12) gene rearrangements.…”

Section: Gene Rearrangement Studiesmentioning

confidence: 99%

Histiocytic sarcoma: a study of five cases including the histiocyte marker CD163

et al. 2005

View full text Add to dashboard Cite

Section: Gene Rearrangement Studiesmentioning

confidence: 99%

Histiocytic sarcoma: a study of five cases including the histiocyte marker CD163

et al. 2005

View full text Add to dashboard Cite

“…Immunoglobulin heavy chain (IgH) gene rearrangement studies were performed by PCR from formalinfixed paraffin-embedded tissue using primers to the V H -FRIII/J H region as previously described 17 and amplified products were separated by capillary electrophoresis with ABI 3100 (Applied Biosystems, Foster City, CA, USA).…”

Section: Gene Rearrangement Studiesmentioning

confidence: 99%

t(11;18)(q21;q21) in extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue in stomach: a study of 48 cases

et al. 2009

View full text Add to dashboard Cite

Gastric extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MZL-MALT) is speculated to be immune mediated and is notable for responding to treatment by Helicobacter pylori eradication. However, the gastric MZL-MALT with t(11;18)(q21;q21) has been shown to be resistant to treatment by H. pylori eradication. We studied the molecular, immunohistochemical, and histological aspects of 48 cases of gastric MZL-MALT and used a reverse transcription real-time PCR assay to assess the presence of a t(11;18)(q21;q21) in formalin-fixed, paraffin-embedded tissue. Florescence in situ hybridization for t(11:18)(q21;q21) was used to confirm the real-time PCR results. Three distinct morphological subtypes were recognized: monocytoid, small lymphocytic, and plasmacytoid. Morphology, immunophenotype, and immunoglobulin heavy chain (IgH) gene rearrangement were correlated with the results of the t(11:18)(q21;q21) assay. Of the 48 analyzed cases, 15 (31%) were positive for t(11;18)(q21;q21) and 33 (69%) were monoclonal for IgH gene rearrangement. Of the 15, 13 (87%) cases with t(11;18)(q21;q21) translocation showed IgH gene rearrangement by PCR. Of the 33 t(11;18)(q21;q21)-negative cases tested, 20 cases (61%) showed IgH gene rearrangement. The 15 t(11;18)(q21;q21) translocation-positive cases had either monocytoid (12 of 15) or small lymphocytic morphology (3 of 15). Aberrant expression of CD43 was observed in 8 of 15 (53%) t(11;18)(q21;q21)-positive cases and 21 of 31 (68%) t(11;18)(q21;q21)-negative cases. Our data show that t(11;18)(q21;q21)-positive MZL-MALTs frequently show monocytoid morphology, less often small lymphocytic morphology, and not purely plasmacytoid morphology. Identification of a t(11;18)(q21;q21) by reverse transcription real-time PCR is highly specific for MZL-MALT and helps in the diagnosis of MZL-MALT. Studying the correlation between this translocation and morphological features may increase our understanding of the role of this translocation in the pathogenesis and the clinical behavior of gastric MZL-MALT.

show abstract

“…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.…”

Section: Polymerase Chain Reaction (Pcr)-based Analysis For Detectingmentioning

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

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

Determination of B-Cell Clonality in Paraffin-Embedded Lymph Nodes Using the Polymerase Chain Reaction

Cited by 83 publications

References 0 publications

Histiocytic sarcoma: a study of five cases including the histiocyte marker CD163

Histiocytic sarcoma: a study of five cases including the histiocyte marker CD163

t(11;18)(q21;q21) in extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue in stomach: a study of 48 cases

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

Contact Info

Product

Resources

About