Immunophenotypic evaluation of the Bone Marrow in non-Hodgkin's lymphoma

doi:10.1016/0046-8177(93)90244-B

Human Pathology

Volume 24, Issue 6, June 1993, Pages 636-642

https://doi.org/10.1016/0046-8177(93)90244-B Get rights and content

Abstract

Immunophenotypic evaluations of the bone marrow (BM) are reported on 69 aspirates from 58 patients who had non-Hodgkin's lymphoma or chronic lymphocytic leukemia involving the BM. Using flow cytometry and immunofluorescence microscopy on density gradient isolated BM mononuclear cells, the neoplasm could be identified and characterized in 59 aspirates (86%) from 49 patients (84%). Using International Working Formulation guidelines the neoplasms were classified on the basis of prior or subsequent histopathology of lymph node, spleen, skin, or other soft tissue site, or by evaluation of peripheral blood in chronic lymphocytic leukemia. In nine cases the lymphoma could not be completely classified according to International Working Formulation guidelines because only BM was available for evaluation. The neoplasm in the BM was identified and characterized immunophenotypically in all 29 cases of chronic lymphocytic leukemia/well-differentiated lymphocytic lymphoma (WDLL) (100%), in 11 of 12 cases of low-grade lymphoma other than WDLL (92%), in 11 of 15 cases of intermediate-grade lymphoma (73%), and in two of four cases of high-grade lymphoma (50%). Six of the nine cases not classified by International Working Formulation guidelines could be characterized immunophenotypically. In 10 cases immunophenotypic studies revealed negative findings, although the concurrent core biopsy specimens were positive. In two cases immunophenotypic studies with positive findings accompanied a negative core biopsy specimen. A panel of immunohistochemical reagents reactive with fixative/paraffin-resistant antigens was used for a retrospective evaluation of the 69 core biopsy specimens. When compared with the immunophenotypic data obtained from the marrow aspirates these results proved to be only moderately reliable in B-lineage neoplasmas and unreliable in T-cell neoplasms. Thus, immunophenotyping of aspirated marrow by flow cytometry was found to be the most reliable method for determining the antigenic profiles of BM-based lymphomas.

References (20)

GS Wood et al.
The immunologic phenotyping of bone marrow biopsies and aspirates: Frozen section techniques
Blood
(1982)
P Clark et al.
Lymphocyte subsets in normal bone marrow
Blood
(1986)
KA Foon et al.
Immunologic classification of leukemia and lymphoma
Blood
(1986)
K Foucar
Bone marrow examination in the diagnosis of acute and chronic leukemias
Hematol Oncol Clin North Am
(1988)
SS Shin et al.
Immunoarchitecture of normal human bone marrow: A study of frozen and fixed tissue sections
Hum Pathol
(1992)
LJ Picker et al.
Immunophenotypic criteria for the diagnosis of non-Hodgkin's lymphoma
Am J Pathol
(1987)
M Chilosi et al.
Routine immunofluorescent and histochemical analysis of bone marrow involvement of lymphoma/leukaemia: The use of cryostat sections
Br J Cancer
(1983)
HD Feiner et al.
IgM monoclonal gammopathy/Waldenstrom's macroglobulinemia: A morphological and immunophenotypic study of the bone marrow
Mod Pathol
(1990)
H Thaler et al.
Bone marrow diagnosis in lymphoproliferative disorders: Comparison of results obtained from conventional histomorphology and immunohistology
Histopathology
(1991)
H Mullink et al.
Influence of fixation and decalcification on the immunohistochemical staining of cell-specific markers in paraffin-embedded human bone marrow biopsies
J Histochem Cytochem
(1985)

There are more references available in the full text version of this article.

Cited by (26)

Systematic review of staging bone marrow involvement in B cell lymphoma by flow cytometry
2021, Blood Reviews
Citation Excerpt :
The latter is particularly seen when BMI is paratrabecular, which is most common in FL [19,28,36,40–42]. Conversely, sampling-related false negatives in DLBCL and other large cell lymphomas may be more likely secondary to their greater cellular fragility and tissue necrosis [17,43–45], as these have been reported as common sources of false negatives in lymph node samples [46–48]. Lack of observer experience or proficiency can also lead to false negative results of FC assessment of BMI, and a number of potential pitfalls should be borne in mind, even when an optimal antibody tube or panel is used.
The clinical relevance of flow cytometry (FC)-based bone marrow involvement (BMI) in B cell non-Hodgkin lymphoma (B-NHL) is not well established. We conducted a systematic review of MEDLINE regarding the use of FC to establish BMI in B-NHL to determine the prevalence of BMI by FC, to understand the interrelation between FC and bone marrow biopsy (BMB), and to explore the prognostic impact of BMI by FC. Relevant exclusion criteria included publication before 2010. Eleven publications (of 18 screened) were included, with 2803 patients involved. Relevant methodological details were often unreported. The prevalence of BMI by FC varied based on histological subtypes included. The median kappa agreement between BMB and FC was 0.68 and the type of discordance (FC+/BMB- vs. FC-/BMB+) was highly variable across studies. Only 4 studies (all in diffuse large B cell lymphoma) assessed the prognostic impact of BMI by FC. Two found a worse prognosis for patients with FC+/BMB- than those without BMI. To conclude, studies assessing BMI by FC are retrospective, of low methodological quality and with heterogeneous findings.
Bone marrow
2009, Modern Surgical Pathology
Bone Marrow
2009, Modern Surgical Pathology
Magnetic resonance imaging of bone marrow versus bone marrow biopsy in malignant lymphoma
1999, Pathology and Oncology Research
Bone marrow involvement is a frequent finding in malignant lymphoma. Bone marrow biopsy of the posterior iliac crest is routinely performed for staging. Abnormal magnetic resonance imaging (MRI) signals of bone marrow was also reported to be indicative of bone marrow involvement. This study included 60 patients with malignant lymphoma. Unilateral bone marrow biopsy of the posterior iliac crest was performed. MRI of lumbar spine was studied within 24 hours of bone marrow biopsy. 22 healthy controls were used for the detection of MRI objectivity during visual evaluation. In 83% of patients (50/60), biopsy and MRI results agreed completely. In two patients, histologic sections failed to show any evidence of bone marrow involvement despite abnormal MRI signals suggestive of involvement. In three patients, MRI was completely normal despite biopsy proven bone marrow infiltration. False negativity (3/60) and false positivity (2/60) rates were very low. Negative biopsy findings with positive or equivocal MRI results should not exclude bone marrow involvement and need further evaluation with bilateral or guided biopsy. Thus, we conclude that MRI of bone marrow is a fairly sensitive, noninvasive modality and might be of potential value in detecting bone marrow infiltration in malignant lymphoid neoplasms which can be utilized as a useful adjunct to standard staging procedures. Copyright 1999 W.B. Saunders Company Ltd on behalf of the Arányi Lajos Foundation
Cold acetone fixation and methacrylate embedding. A suitable method for routine processing of bone marrow biopsies
1998, Pathology Research and Practice
Here we report that acetone fixation at −18 °C with subsequent embedding in methyl-/butylmethacrylate is a reliable method for the routine processing of bone marrow biopsies. This method allows good conventional histological visualization of morphological details, which is comparable with other fixation procedures. The essential advantage of this method is that a wide range of monoclonal antibodies and polyclonal antisera can be used for immunohistochemical investigations for diagnostic and scientific purposes. The addition of 5% polyethylene glycol 400 to the acetone minimizes freeze-related artefacts. The immunohistochemical demonstration of a number of antigens is mostly affected by the medium used for slide preparation and to a lesser extent by the concentration of benzoylperoxide used for polymerization. Performing polymerization at 4 °C and using N, N-dimethyl-p-toluidine as accelerator allows the concentration of benzoylperoxide to be reduced to 0.2 g% (8.3 mmol). Under these conditions the methacrylate embedding procedure has only minimal effects on the quality of immuno- and enzyme histochemistry. Additionally, the simplified method for removing the polymerization inhibitor from the methacrylate components and the shortened impregnation step are further advantages of the embedding method described here.
Bone marrow one step fixation-decalcification in Lowy FMA solution: An immimohistological and in situ hybridization study
1994, Pathology Research and Practice
The immunoreactivity of paraffin embedded bone marrow biopsies (BMB) was studied following a one step 20-hour-fixation-decalcification in Lowy formalin mercuric chldrid acid solution wbich permits excellent histological stainings.
Antibodies reactive with myeloid, megakaryocyte, erythroid cells, T and B lymphocytes, mastocytes and metastatic cells were compared. Nearly all antibodies working on paraffin sections were demonstrated on Lowy FMA fixed BMB. Special care was taken to define an optimal working dilution. Trypsinization was not necessary. A slide microwave pre-treatment appeared essential before testing CD20 L26, CD8, CD3, CD34, MB1 Kappa and Lambda antibodies. It was suitable for UCHL1, LN2, CD30 antibodies.
The same fixative allowed an m RNA Kappa or Lambda in myeloma and EBER 1 EB V RNAs in HIV lymphoma visualization by in situ hybridization. The safety handling of the toxic mercuric chloride component is discussed.

View all citing articles on Scopus

^☆: Presented in part at the 72nd Annual Meeting of the US and Canadian Academy of Pathology, Chicago, IL, March 1991.

View full text

Original contributionImmunophenotypic evaluation of the Bone Marrow in non-Hodgkin's lymphoma☆

Abstract

Blood

Blood

Blood

Hematol Oncol Clin North Am

Hum Pathol

Immunophenotypic criteria for the diagnosis of non-Hodgkin's lymphoma

Am J Pathol

Routine immunofluorescent and histochemical analysis of bone marrow involvement of lymphoma/leukaemia: The use of cryostat sections

Br J Cancer

IgM monoclonal gammopathy/Waldenstrom's macroglobulinemia: A morphological and immunophenotypic study of the bone marrow

Mod Pathol

Bone marrow diagnosis in lymphoproliferative disorders: Comparison of results obtained from conventional histomorphology and immunohistology

Histopathology

Influence of fixation and decalcification on the immunohistochemical staining of cell-specific markers in paraffin-embedded human bone marrow biopsies

J Histochem Cytochem

Original contribution
Immunophenotypic evaluation of the Bone Marrow in non-Hodgkin's lymphoma☆