The detection of B-cell monoclonal populations by polymerase chain reaction: Accuracy of approach and application in gastric endoscopic biopsy specimens

doi:10.1016/0046-8177(93)90214-2

Human Pathology

Volume 24, Issue 11, November 1993, Pages 1184-1188

https://doi.org/10.1016/0046-8177(93)90214-2 Get rights and content

Abstract

The recently developed strategy for the detection of monoclonal B-cell populations, based on the selective amplification of predominant immunoglobulin heavy chain gene (IgH) rearrangement, is seen to be a suitable alternative to Southern blot analysis. The new technique uses a pair of consensus primers for variable (V_H) and joining (J_H) regions. We first tested the accuracy of this new approach on a broad series of 67 samples that had been well characterized by both Southern blot and immunohistochemical techniques before being subjected to blind testing. Our results show that this technique gave 100% specificity (absence of false-positive results) and approximately 70% sensitivity (30% false-negative results). The only exception was the presence of an IgH polymerase chain reaction ambiguous result in a case of Sezary's syndrome. The polymerase chain reaction technique was then applied to a panel of 27 frozen gastric endoscopy biopsy specimens following previous clinical suspicion of lymphoma. Monoclonality was detected in nine of 13 samples previously diagnosed as lymphomas and in one of two carcinomas. Further examination of the gastrectomy specimen in the latter case disclosed a B-cell lymphoma associated with the carcinoma. In spite of its limited sensitivity, the high specificity attained by this technique in the detection of monoclonality makes it a useful adjunct to routine morphologic criteria, as it is sometimes capable of detecting true positive cases that conventional morphology studies show to be negative.

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Cited by (39)

Helicobacter pylori-negative low-grade extranodal B-cell primary gastric mucosa-associated lymphoid tissue lymphoma
2007, Journal of the Chinese Medical Association
Helicobacter pylori-negative primary mucosa-associated lymphoid tissue (MALT) lymphoma of the stomach-a primary gastric lymphoma (PGL)-is not very common. Herein, we report a 77-year-old man with the chief complaint of intermittent epigastric pain. Initial upper gastrointestinal (UGI) endoscopy showed multiple irregular ulcers (GUs) over the antrum with poor distention of the stomach, and gastric adenocarcinoma (Borrmann type IV, linitis plastica) was suspected. But the biopsy specimens of the GUs showed chronic inflammation without malignant change or H. pylori infection. Nevertheless, delayed healing of the GUs, specific endoscopic findings and enlarged paragastric lymph nodes on computed tomography of the abdomen alerted us to the possibility of PGL. After repeat UGI endoscopies, the patient was finally diagnosed with extranodal B-cell PGL. The patient received radical subtotal gastrectomy and Billroth II operation; the surgical margins were free of tumor cells. The patient was followed up in the outpatient department for 20 months, and his postoperative condition was good.
Gastric mucosa-associated lymphoid tissue lymphoma
2003, American Journal of Gastroenterology
Citation Excerpt :
Brush cytology may provide some additional benefit (47). Other methods to improve the diagnostic yield of endoscopic biopsy specimens include the use of immunohistochemical techniques (48), as well as PCR assays to detect monoclonal B cell populations (49). Since the introduction of these techniques, problems in differentiating high grade lymphoma from adenocarcinoma have largely been solved.
The connection between Helicobacter pylori and gastric mucosa-associated lymphoid tissue (MALT) lymphoma is well established. H. pylori infection causes an immunological response, leading to chronic gastritis with formation of lymphoid follicles within the stomach. These lymphoid follicles resemble nodal tissues found throughout the body and are composed of reactive T cells and activated plasmal cells and B cells. The B cells are responsible for initiating a clonal expansion of centrocyte-like cells that form the basic histology of MALT lymphoma. Early diagnosis of MALT lymphoma is difficult but essential for adequate treatment. Clinical symptoms are vague and varied, with abdominal pain being a common presenting complaint. The endoscopic appearance of this tumor is varied and can be infiltrative, exophytic, or ulcerative. In addition, the tumor can have a multifocal distribution, and therefore aggressive tissue sampling is crucial for diagnosis. Endoscopic ultrasound is essential to document the extent of disease and is more accurate than CT scan in detection of spread to perigastric lymph nodes. Lesions that are confined to the mucosa or submucosa of the gastric wall are believed to be dependent on H. pylori stimulation and therefore can be successfully treated with H. pylori eradication. Those MALT lymphomas that present at more advanced stages require more aggressive management and can be treated with surgical resection, radiation, or chemotherapy. Follow-up is critical in all patients who have been treated with H. pylori eradication and consists of multiple endoscopic biopsies for histological and molecular studies as well as endoscopic ultrasound at 3, 6, and 12 months after treatment. The reappearance of MALT lymphomas has been seen years after treatment, and therefore follow-up of these patients should be indefinite.
Molecular diagnostics in low-grade gastric marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue type after Helicobacter pylori eradication therapy
2001, Clinical Lymphoma
The primary gastric lymphomas are extranodal non-Hodgkin's lymphomas that likely originate from the mucosa-associated lymphoid tissue (MALT). Data suggest that chronic infection with Helicobacter pylori (H pylori) is significantly associated with the pathogenesis of low-grade gastric MALT lymphomas. This is in keeping with the observation that many patients with early low-grade MALT lymphomas have complete remissions after H pylori eradication therapy. However, the stability of these remissions remains unclear and relapses have been reported. It can be difficult to distinguish between early malignant and benign disorders of the gastric mucosa. A polymerase chain reaction (PCR) assay can detect rearrangements of the variable region of immunoglobulin heavy chains. This assay can be used to distinguish the clonality of B lymphocytes and has been investigated as a test for differential diagnosis of MALT lymphomas. Monoclonality is observed in the majority of MALT-lymphoma samples at diagnosis but has been found in gastritis samples as well. Whether the presence of monoclonal B cells is associated with the risk of lymphoma progression remains unclear. As many as 50% of patients who have complete histologic remissions of MALT lymphoma after H pylori eradication therapy have persisting monoclonal bands in follow-up PCR monitoring. Although it is unclear as to whether monoclonality indicates the presence of minimal residual disease, patients who have persistent monoclonal bands during follow-up should be considered at risk for relapse. The PCR assay for rearrangements of the variable region of the immunoglobulin heavy-chain gene appears to be of low value in the diagnosis of B-cell malignancies but could provide a useful tool in the follow-up of patients who achieve remissions after H pylori eradication.
Low prevalence of monoclonal B cells in Helicobacter pylori gastritis patients with duodenal ulcer
1998, Human Pathology
We have studied the prevalence of B-cell clonality among a large group of 320 patients with Helicobacter pylori gastritis and duodenal ulcer. These patients underwent endoscopic examination with multiple gastric biopsies at diagnosis and were followed 2 and 12 months after therapy. Histopathologic examination of 809 sets of biopsy specimens showed lymphoid gastritis with lymphoid aggregates or follicles, but without lymphoepithelial lesion, in 302 samples corresponding to initial biopsy specimens (n = 130) or to posttreatment biopsy specimens (n = 172). DNA extracted from fresh antral specimens allowed the amplification of Helicobacter pylori DNA in all cases before therapy. The arrangement of the immunoglobulin heavy chain gene was studied by polymerase chain reaction (PCR) in the 302 selected lymphoid gastritis samples. Single or dominant bands were seen only in four specimens from three patients (1.3%), whereas a polyclonal pattern was seen in the other 298 samples. The detection threshold of our PCR technique was approximately 3% of clonal B cells diluted in a polyclonal population. This threshold appeared to be a reliable cutoff between polyclonal gastritis and clonal MALT lymphoma. In our experience, Helicobacter pylori lymphoid gastritis appeared mainly as a benign polyclonal condition.
The value of EUS in combination with cytological, flow cytometry, and gene rearrangement in the diagnosis of gastrointestinal lymphoma
2017, Hematological Oncology
Clonal immunoglobulin heavy chain and T-cell receptor γ gene rearrangements in primary gastric lymphoma
2013, World Journal of Gastroenterology

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^☆: Supported by a grant from the Fondo de Investigaciones Sanitarias (FIS), Spain.

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Original contributionThe detection of B-cell monoclonal populations by polymerase chain reaction: Accuracy of approach and application in gastric endoscopic biopsy specimens☆

Abstract

Blood

Anal Biochem

Blood

Immunol Today

Hum Pathol

Immunoglobulin gene rearrangements as unique clonal markers in human lymphoid neoplasms

N Engl J Med

Somatic generation of antibody diversity

Nature

Detection of minimal disease in hematopoietic malignancies of the B-cell lineage by using third-complementarity-determining region (CDR-III)-specific probes

Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase

Science

Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes), and Fast red as chromogen

J Histochem Cytochem

The Kiel classification

Original contribution
The detection of B-cell monoclonal populations by polymerase chain reaction: Accuracy of approach and application in gastric endoscopic biopsy specimens☆