6
views
0
recommends
+1 Recommend
1 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      A importância da citometria de fluxo no diagnóstico raro de mieloma mielomonocítico Translated title: The importance of flow cytometry in the rare diagnosis of myelomonocytic myeloma

      case-report

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          O mieloma múltiplo consiste na proliferação de células plasmáticas. Raramente apresenta, ao diagnóstico, morfologia de células imaturas com citoplasma amplo basofílico, sugerindo quadro leucêmico inicial. O objetivo deste artigo é mostrar a importância da imunofenotipagem na elucidação destes achados morfológicos pela expressão simultânea de antígenos plasmocíticos, mielomonocíticos e de linhagem linfóide T, confirmando a hipótese diagnóstica de mieloma mielomonocítico. Apresentamos dois casos de mieloma mielomonocítico através de análise morfológica (coloração pancromática de Romanovsky), citoquímica (PAS, peroxidase, sudan black, alfanaftil acetato esterase e oil red), citogenética e imunofenotipagem por citometria de fluxo em sangue periférico e medula óssea, de acordo com as técnicas recomendadas. Foram utilizados os anticorpos monoclonais: CD2, CD3, CD4, CD5, CD7, CD10, CD13, CD15, CD19, CD20, CD25, CD33, CD34, CD38, CD45, CD56, CD71, HLA-DR, TCR alfa/beta, TCR gama/delta, kappa, lambda, IgM e IgD de superfície e intracitoplasmática, assim como MPO, CD79a e CD3 intracitoplasmático. Utilizamos as técnicas de banda G e FISH nas análises citogenéticas. Foram observadas alterações clonais em ambos os casos, sendo uma com trissomia do cromossomo 8 e outro caso com deleção do braço longo do cromossomo 7 e do braço curto do cromossomo 6. Os percentuais de positividade encontrados nos anticorpos monoclonais CD4, CD7, CD10, CD13, CD14, CD15, CD33, CD38 e CD56 de forte expressão, HLA-DR, TCR gama/delta, MPO e IgM intracitoplasmático no histograma de volume x complexidade e no histograma de CD45 x complexidade permitiram concluir este diagnóstico em ambos os casos, demonstrando a importância do método.

          Translated abstract

          Multiple myeloma is a malignant disease caused by unregulated proliferation of mature plasma cells. Very few cases present as blast cells in the bone marrow with intense basophylic cytoplasm and monocytoid morphology, suggesting acute leukemia. This paper presents immonophenotypying in 2 cases with the diagnostic hypoteses of myelomonocytic myeloma, showing the simultaneous expression of plasmocytic, myelomonocytic and T cell antigens. Immonophenotypying was done in peripheral blood and bone marrow with classical techniques, using the following monoclonal antibodies: CD2, CD3, CD4, CD5, CD7, CD10, CD19, CD20, CD25, CD33, CD34, CD38, CD41, CD45, CD56, CD71, HLA-DR, TCR alpha/beta, DCR gama/delta, kappa, lambda, surface IGM and IGG, intracitoplasmic kappa, lambda, IGM and IGD, MPO, CD79a and intracitoplasmic CD3. Morphology was evaluated with pan-chromatic Romanovsky stain and cytochemical stains - PAS, peroxidase, Sudan Black B, alpha-naphtil acetate esterase and Oil Red O. Cytogenetic analysis was done with banding techniques and FISH. Clonal cytogenetic alterations were present in both cases, one with chromosomo 8 trysomy and the other with long arm delection of chromosome 7 and short arm delection of chromosome 6. The percentage positivity of specific markers were clean cut making the diagnosis clear.

          Related collections

          Most cited references19

          • Record: found
          • Abstract: found
          • Article: not found

          Immunophenotypic characterization of plasma cells from monoclonal gammopathy of undetermined significance patients. Implications for the differential diagnosis between MGUS and multiple myeloma.

          Although the immunophenotype of plasma cells (PCs) from multiple myeloma (MM) patients has been extensively explored, information on the phenotypic characteristics of PCs in monoclonal gammopathy of undetermined significance (MGUS) patients is scanty and frequently controversial. Thus, the question of whether or not PCs are phenotypically different in the two disorders and whether this criteria could be useful for the differential diagnosis between MGUS and MM remains to be explored. In the present study, the immunophenotypic profile of bone marrow PCs (BMPCs) from a group of 76 MGUS patients has been analyzed by flow cytometry and compared with that of BMPCs present in both MM patients (n = 65) and control subjects (n = 10). For that purpose, a large panel of monoclonal antibodies against PC-related antigens was used together with a sensitive methodology in which a minimum of 10(3) PCs were studied. In all MGUS cases studied, two clearly defined and distinct PC subpopulations could be identified. One PC subpopulation, population A (33 +/- 31% of total PCs), constantly displayed a high CD38 expression with low forward light scatter (FSC)/side light scatter (SSC) and was positive for CD19 and negative for CD56 (only a small proportion of these PCs were weakly positive for CD56). The other PC subpopulation, population B (67 +/- 31% of total PCs), showed the opposite pattern; the antigen CD56 was strongly positive and CD19 was constantly negative, and it showed a lower CD38 expression and higher FSC/SSC values than population A. Clonality studies (cytoplasmic light chain restriction, DNA content studies, and polymerase chain reaction assessment) confirmed the clonal nature of PCs from population B and the polyclonal origin of PCs from population A. Moreover, the polyclonal PCs from MGUS displayed a phenotypic profile identical to that found in PCs from healthy individuals. By contrast, clonal PCs from all MGUS patients displayed a similar antigenic profile to myelomatous PCs, with clear phenotypic differences with respect to normal PCs: lower intensity of CD38 expression and a variable reactivity for markers that were not expressed in normal PCs, such as CD28, CD117, and sIg. Although the presence of residual polyclonal PCs was a constant finding in MGUS patients, it was a rare event in MM and, when present (only 22% of MM cases), its frequency was significantly lower than that observed in MGUS (0.25% versus 32.9%, respectively; P < 0.0001). Only 1.5% of patients with MM had more than 3% of normal PCs, whereas 98% of patients with MGUS had more than 3%. Moreover, as shown by multivariate analysis, the number of residual polyclonal PCs was the most powerful single parameter for the discrimination between MGUS and MM patients at diagnosis, even when only stage I MM cases were considered.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Cell surface markers in multiple myeloma.

            To describe the immunophenotype of normal and myelomatous plasma cells (PCs) and the changes in immunoregulatory nonmyelomatous cells in multiple myeloma (MM). The cell surface markers (antigens) associated with this common cancer were reviewed. Immunophenotypic characterization of both normal PCs and their counterpart malignant hematopoietic cells can be achieved by using monoclonal antibodies and either flow cytometry or immunocytochemical techniques. Normal PCs are heterogeneous and express, in addition to cytoplasmic immunoglobulins, the antigens CD9, CD10, CD13, CD19, CD20, CD33, CD38, and D-related human leukocyte antigen (HLA-DR). This heterogeneity also occurs in malignant PCs. Myelomatous PCs may express, in addition to CD38 (the most typical PC marker), the antigens CD9, CD10, HLA-DR, and CD20. Other non-B-cell lineage markers such as myeloid (CD13, CD14, CD15, CD33, CD41, and glycophorin A), T-cell (CD2 and CD4), and natural killer-associated (CD56) antigens, as well as CD23, CD24, CD25, CD37, CD39, CDw40, CD45R, CD71, and certain unclustered antigens (R1-3, PCA-1, PCA-2, PC1, 62B1, 8A, 8F6, and MM4), have been noted in myelomatous PCs. The presence of these antigens in the myeloma cells may have a prognostic value--for example, the expression of CD20 and of myelomonocytic antigens (CD11b, CD13, CD14, CD15, and CD33) may be related to a poor prognosis. The adverse prognostic implication of the expression of CD10 initially described in MM has not been subsequently confirmed. Patients with MM may have mononuclear cells in their peripheral blood that express the same antigens as those expressed by the myeloma cells in their bone marrow. The presence of such cells or their therapy-associated decrease or disappearance may be related to the prognosis of patients with MM. The presence of cell surface markers on PCs and their prognostic significance in patients with MM warrant further investigation.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              Characterization of aberrant phenotypes in acute myeloblastic leukemia.

              The existence of leukemic-associated phenotypes has been suggested to be a valuable tool for the detection of minimal residual disease (MRD) in AML patients, as they would allow to distinguish leukemic blast cells from normal hematopoietic progenitors. The present study was designed to analyze in which proportion of AML patients the immunological detection of MRD is feasible, based on the presence of aberrant phenotypes that allow the distinction of leukemic from normal cells. For this purpose we have prospectively investigated the blast cells from 40 AML patients at diagnosis with a large panel of MoAb in double and triple staining combinations analyzed at flow cytometry, in order to detect aberrant phenotypes on blast cells (lineage infidelity, antigenic overexpression, and asynchronous antigenic expression, as well as aberrant light-scatter pattern). In the analysis of the 40 AML cases more than one blast cell subset, distinguished by its different antigenic expression, was detected in 85% of the patients: five different phenotypic blast cell subsets were observed in six cases, four in 13 patients, three subsets in three cases, and two in 12 patients; only six cases showed a homogeneous phenotypical blast cell population. Twenty-nine of the 40 AML cases analyzed (73%) showed the existence of at least one aberrant phenotype: in 15 cases the myeloid blast cells co-expressed lymphoid-associated antigens (CD2, CD5, CD7, and/or CD19)--lineage infidelity--; asynchronous antigen expression was detected in 25 patients (CD34+CD56+, CD34+CD11b+, CD34+CD14+, CD117+CD15+, CD33-CD13+, CD13-CD15+, HLADR + CD15 , HLADR-CD14+CD11b+ CD4+); seven cases displayed antigen overexpression (CD13, CD33, CD15, or CD14); and in 13 patients leukemic cells had an abnormal FSC/SSC distribution according to their phenotype. These results suggest that immunological methods for the detection of MRD based on the existence of aberrant phenotypes could be used in the majority of AML patients.
                Bookmark

                Author and article information

                Journal
                jbpml
                Jornal Brasileiro de Patologia e Medicina Laboratorial
                J. Bras. Patol. Med. Lab.
                Sociedade Brasileira de Patologia Clínica (Rio de Janeiro, RJ, Brazil )
                1676-2444
                1678-4774
                January 2002
                : 38
                : 1
                : 25-31
                Affiliations
                [02] orgnameHIAE
                [07] orgnameHIAE orgdiv1Laboratório de Técnicas Especiais
                [04] orgnameHIAE orgdiv1Laboratório Clínico
                [09] orgnameSanta Casa de São Paulo orgdiv1Faculdade de Ciências Médicas
                [01] orgnameHospital Albert Einstein (HIAE) orgdiv1Setor de Citometria de Fluxo
                Article
                S1676-24442002000100006 S1676-2444(02)03800106
                d7b5c850-ad50-4ad6-b29c-a20178ff8049

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 01 October 2001
                : 06 March 2001
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 19, Pages: 7
                Product

                SciELO Brazil

                Categories
                Medicina Laboratorial

                Flow cytometry,Cytogenetic analysis,Morphologic analysis,Multiple myeloma,Citogenética,Citometria de fluxo,Análise morfológica,Mieloma mielomonocítico

                Comments

                Comment on this article