Automated kappa and lambda light chain mRNA expression for the assessment of B‐cell clonality in cutaneous B‐cell infiltrates: its utility and diagnostic application

Magro, Cynthia M.; Crowson, A. Neil; Porcu, Pierluigi; Nuovo, Gerard J.

doi:10.1034/j.1600-0560.2003.00102.x

Cited by 32 publications

(51 citation statements)

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“…To detect B cells/plasma cells, we stained tissue specimens for Ig kappa light chain, which was shown to be a reliable marker for B cells and terminally differentiated Ab-secreting plasma cells (30, 38). As a positive control, we used human tonsil specimens, which revealed the presence of immunopositive cells within the sub-epithelial layer most likely representing a germinal center (Fig.…”

Section: Resultsmentioning

confidence: 99%

The B Cell–Stimulatory Cytokines BLyS and APRIL Are Elevated in Human Periodontitis and Are Required for B Cell–Dependent Bone Loss in Experimental Murine Periodontitis

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AlSarhan

Benakanakere

et al. 2015

The Journal of Immunology

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B-lineage cells (B lymphocytes and plasma cells) predominate in the inflammatory infiltrate of human chronic periodontitis. However, their role in disease pathogenesis and the factors responsible for their persistence in chronic lesions are poorly understood. In this regard, two cytokines of the TNF ligand superfamily, namely a proliferation-inducing ligand (APRIL) and B-lymphocyte stimulator (BLyS), are important for the survival, proliferation, and maturation of B cells. We thus hypothesized that APRIL and/or BLyS are upregulated in periodontitis and contribute to induction of periodontal bone loss. This hypothesis was addressed in both human and mouse experimental systems. We show that, relative to healthy controls, the expression of APRIL and BLyS mRNA and protein was upregulated in natural and experimental periodontitis in humans and mice, respectively. The elevated expression of these cytokines correlated with increased numbers of B cells/plasma cells in both species. Moreover, APRIL and BLyS partially colocalized with kappa light chain-expressing B lineage cells at the epithelial-connective tissue interface. Ligature-induced periodontitis resulted in significantly less bone loss in B cell-deficient mice compared to wild-type controls. Ab-mediated neutralization of APRIL or BLyS diminished the number of B cells in the gingival tissue and inhibited bone loss in wild-type but not in B cell-deficient mice. In conclusion, B cells and specific cytokines involved in their growth and differentiation contribute to periodontal bone loss. Moreover, APRIL and BLyS have been identified as potential therapeutic targets in periodontitis.

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Section: Resultsmentioning

confidence: 99%

The B Cell–Stimulatory Cytokines BLyS and APRIL Are Elevated in Human Periodontitis and Are Required for B Cell–Dependent Bone Loss in Experimental Murine Periodontitis

Abe

AlSarhan

Benakanakere

et al. 2015

The Journal of Immunology

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“…While many methods can be used, the most common (flow cytometry) requires fresh tissue, or does not take into account morphologic context (PCR), or is insufficiently sensitive for the majority of B cell NHL (current IHC and ISH methods) [5,6]. In this study, we describe the feasibility and application of dual-color CISH technology for the evaluation of clonality of mature B cell populations in tissue sections.…”

Section: Resultsmentioning

confidence: 99%

“…In situ hybridization (ISH) for mRNA is therefore used in many laboratories. However, current methods allow visualization of mRNA levels of B cells only in the later stages of differentiation and so do not have the dynamic range of flow cytometry (see Figure 1) [5,6] Finally, molecular methods aimed at amplification of KAPPA and LAMBDA light chain genes and examination for a predominant rearrangement motif are also used. These techniques usually performed in molecular laboratories, require longer time as nucleic acid extraction and amplification are needed and do not retain morphologic context i.e.…”

Section: Introductionmentioning

confidence: 99%

Kappa and lambda light chain mRNA in situ hybridization compared to flow cytometry and immunohistochemistry in B cell lymphomas

et al. 2014

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BackgroundDetection of B cell clonality is useful for assisting in the diagnosis of B cell lymphomas. Clonality assessment can be accomplished through evaluation of KAPPA and LAMBDA light chain expression. Currently, only slide based methods are available for the majority of patient biopsies and do not detect light chain protein or mRNA in many B-cell lymphomas. Herein we evaluated a new method, known as colorimetric in situ hybridization (CISH), with improved sensitivity and multiplexing capacity, for its usefulness in clonality detection in mature B cell malignancies.MethodsThe KAPPA and LAMBDA ISH was performed on a Ventana Benchmark XT utilizing two color chromogenetic detection. The probes comprised 2 haptenated riboprobes each approximately 500 base pairs long directed against the conserved regions of either KAPPA or LAMBDA mRNA. The dual colors consisted of silver deposition (black) for KAPPA light chain and a novel (pink) chromogen for LAMBDA light chain. Following optimization, CISH allowed visualization of mRNA in benign B cells in reactive tissues including germinal center, mantle zone, and post-germinal center cells. We then identified 79 cases of B cell lymphoma with formalin-fixed paraffin-embedded (FFPE) biopsies including: follicular (36 cases), mantle cell (6 cases), marginal zone (12 cases), lymphoplasmacytic (6 cases), small lymphocytic (4 cases), and diffuse large B cell (15 cases), which were selected on the basis of either prior flow cytometry or immunohistochemistry (IHC) results to serve as the predicate, "gold standard," comparator.Results39/79 (49.4%) cases were classified as KAPPA and 29/79 (36.7%) as LAMBDA light chain restricted; while 9/79 (11.3%) cases were classified as indeterminate. Of the 70 cases with KAPPA or LAMBDA light chain restricted CISH, 69/70 (98.6%) were concordant with the reference method, while 1/70 (1.4%) was discordant.ConclusionsOptimized CISH detected lower levels of mRNA than can be visualized with current slide based methods, making clonality assessment in FFPE biopsies possible for mature B cell neoplasms. In this preliminary study, CISH was highly accurate compared to flow cytometry or IHC. CISH offers the possibility of wider applicability of light chain ISH and is likely to become a useful diagnostic tool.Virtual SlidesThe virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1430491067123856

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“…Das primär kutane Marginalzonenlymphom ist geprägt durch kleine Marginalzonenzellen, die histomorphologisch an Zentrozyten erinnern, sich jedoch immunhistologisch eindeutig abgrenzen lassen: Expression von BCL2 bei Negativität für den Keimzentrumsmaker BCL6. Daneben sind die Infiltrate reich an lymphoplasmazytoiden Zellen und Plasmazellen mit zytoplasmatischem Immunglobulin, oft IgA, und häufig monoklonaler Expression von κ-oder λ-LeichtkettenProteinen [14]. Reaktive Keimzentren kommen vor und lassen sich immunhistologisch mit Antikör-pern gegen CD21-oder Ki-67-Antigen leicht von malignen Keimzentren des primär kutanen Keimzentrumlymphoms abgrenzen.…”

Section: Primär Kutanes Marginalzonenlymphomunclassified

Maligne Lymphome der Haut

et al. 2011

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The diagnosis of primary cutaneous B-cell lymphoma is made based principally on the results of histological investigations and staging. For an exact staging abdominal sonography and chest X-ray examinations and for appropriate clinical symptoms special investigations as well as radiological imaging procedures including PET are indicated in addition to conventional laboratory investigations. For therapy rituximab is normally administered as monotherapy in order to avoid over therapy of indolent lymphoma. Further options are radiotherapy and new approaches with electrochemotherapy as well as pegylated doxorubicin.

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Automated kappa and lambda light chain mRNA expression for the assessment of B‐cell clonality in cutaneous B‐cell infiltrates: its utility and diagnostic application

Abstract: The Ventana kappa/lambda assay is a reliable, quick, and inexpensive way to determine B-cell clonality in cutaneous lymphoid infiltrates in paraffin-embedded formalin-fixed tissue.

Cited by 32 publications

References 29 publications

The B Cell–Stimulatory Cytokines BLyS and APRIL Are Elevated in Human Periodontitis and Are Required for B Cell–Dependent Bone Loss in Experimental Murine Periodontitis

The B Cell–Stimulatory Cytokines BLyS and APRIL Are Elevated in Human Periodontitis and Are Required for B Cell–Dependent Bone Loss in Experimental Murine Periodontitis

Kappa and lambda light chain mRNA in situ hybridization compared to flow cytometry and immunohistochemistry in B cell lymphomas

Maligne Lymphome der Haut

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