Immunophenotypic Analysis of Human Mast Cells by Flow Cytometry

Escribano, Luís; Navalón, Raquel; Núñez, Rosa; Agustín, Beatriz Díaz; Bravo, Pilar

doi:10.1002/0471142956.cy0606s12

Cited by 13 publications

(17 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Multiparameter flow cytometry immunophenotype of BMMC Multiparameter flow cytometry immunophenotypic studies were performed on BM aspirate samples, processed as previously described, 14,16 using either a FACSCalibur (Becton Dickinson--BD, San Jose, CA, USA) or a FACSCanto II flow cytometer (BDB), equipped with the CellQUEST and FACSDiva software programs (BDB), respectively. Four-color, fluorescein isothiocyanate (FITC)/phycoerythrin (PE)/peridinin chlorophyll protein (PerCP)--cyanin 5.5 (Cy5.5)/allophycocyanin, combinations of monoclonal antibodies (MAb) were used to stain BM cells for a broad set of proteins ( Table 1).…”

Section: Materials and Methods Patientsmentioning

confidence: 99%

An immature immunophenotype of bone marrow mast cells predicts for multilineage D816V KIT mutation in systemic mastocytosis

et al. 2011

View full text Add to dashboard Cite

D816V KIT mutation of bone marrow (BM) mast cells (MC) is a common feature to systemic mastocytosis (SM) patients. Nevertheless, occurrence of the KIT mutation in BM cell compartments other than MC is associated with progression to more aggressive forms of the disease and poor outcome in indolent SM (ISM). Here, we assessed the potential association between the immunophenotype of MC and multilineage KIT mutation in the BM of SM patients through the investigation of the flow cytometric protein expression profile (PEP) of bone marrow mast cells (BMMC) from 70 control individuals and 206 SM patients, classified according to the WHO (World Health Organization), and the degree of involvement of BM hematopoiesis by the D816V KIT mutation; additionally, we developed a score-based class prediction algorithm for the detection of SM cases with multilineage mutation. Our results show that aberrant expression of CD25 with a FceRI lo , FSC lo , SSC lo and CD45 lo immature phenotype of BMMC, in the absence of coexisting normal MC in the BM, was associated with multilineage involvement by the D816V KIT mutation, regardless of the diagnostic subtype of the disease (for example, indolent vs aggressive SM), which supports the utility of the immunophenotype of BMMC as a surrogate marker to screen for multilineage KIT mutation in ISM.Leukemia (2012) 26, 951 --958;

show abstract

Section: Materials and Methods Patientsmentioning

confidence: 99%

An immature immunophenotype of bone marrow mast cells predicts for multilineage D816V KIT mutation in systemic mastocytosis

et al. 2011

View full text Add to dashboard Cite

show abstract

“…The CELLQuest PRO (BD Biosciences) and Paint-A-Gate PRO software programs (BD Biosciences) were used for data acquisition and analysis as described elsewhere. 11,24,28,29 KIT D816V mutation and other KIT mutations localized at codons 814-819 (exon 17) were detected on highly purified (Z97% purity) bone marrow cell populations, as previously described. 27,30 In turn, identification of KIT mutations at exons 2, 9, 10, 11, 13, 14, and 15 was performed on genomic DNA by direct sequencing of the amplified PCR products in both directions, using the dye-deoxy terminator method, in an ABI Prism 3100 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA).…”

Section: Diagnostic Work-up For Mastocytosismentioning

confidence: 99%

Evaluation of the WHO criteria for the classification of patients with mastocytosis

et al. 2011

Self Cite

View full text Add to dashboard Cite

Diagnosis and classification of mastocytosis is currently based on the World Health Organization (WHO) criteria. Here, we evaluate the utility of the WHO criteria for the diagnosis and classification of a large series of mastocytosis patients (n ¼ 133), and propose a new algorithm that could be routinely applied for refined diagnosis and classification of the disease. Our results confirm the utility of the WHO criteria and provide evidence for the need of additional information for (1) a more precise diagnosis of mastocytosis, (2) specific identification of new forms of the disease, (3) the differential diagnosis between cutaneous mastocytosis vs systemic mastocytosis, and (4) improved distinction between indolent systemic mastocytosis and aggressive systemic mastocytosis. Based on our results, a new algorithm is proposed for a better diagnostic definition and prognostic classification of mastocytosis, as confirmed prospectively in an independent validation series of 117 mastocytosis patients.

show abstract

“…These specific antigen stainings, when associated with light scatter parameters, allow for the specific and sensitive identification of MCs in a tissue sample, even when their frequency is as low as 10 Ϫ4 to 10 Ϫ5 (one MC among 10,000 to 100,000 cells) provided that an appropriate combination of fluorochrome-conjugated monoclonal antibodies is used in a direct immunofluorescence assay (11,14). Further, MCs display strong CD33 expression and they are Fc⑀RI ϩ and CD34 Ϫ ; whenever necessary, these markers may improve the identification of CD117 high , CD45…”

Section: Panels Of Fluorochrome-conjugated Monoclonal Antibody Reagentsmentioning

confidence: 99%

“…low MCs in BM and other tissues (11,14). However, simultaneous staining for additional markers is required on most occasions to clearly distinguish between normal and pathologic MCs in mastocytosis, which typically restricts the use of more than two antibody reagents (CD117 and CD45) for the identification of MC.…”

Section: Panels Of Fluorochrome-conjugated Monoclonal Antibody Reagentsmentioning

confidence: 99%

See 1 more Smart Citation

Immunophenotypic analysis of mast cells in mastocytosis: When and how to do it. Proposals of the Spanish Network on Mastocytosis (REMA)

Escribano¹,

Díaz‐Agustín²,

López

et al. 2004

Cytometry Part B Clinical

Self Cite

136

131

View full text Add to dashboard Cite

Background: Mastocytosis is a term used for a heterogeneous group of disorders characterized by an abnormal proliferation and accumulation of mast cells (MCs) in one or multiple tissues including skin, bone marrow, liver, spleen, and lymph nodes, among others.Methods: In recent years, multiparameter flow cytometric studies have shown that pathologic MCs from patients with mastocytosis display unique aberrant immunophenotypic characteristics as compared with normal MCs.Results: Among other features, pathologic MCs show aberrant expression of CD25 and CD2 antigens and abnormally high levels of the CD11c and CD35 complement receptors, the CD59 complement regulatory molecule, the CD63 lysosomal membrane antigen, and the CD69 early-activation antigen. In addition, MCs from mastocytosis express abnormally low levels of CD117 and unexpectedly high light scatter and autofluorescence characteristics.Conclusions: These aberrant immunophenotypic features are of great relevance for the assessment of tissue involvement in mastocytosis with consequences in the diagnosis, classification, and follow-up of the disease and in its differential diagnosis with other entities. In this paper we provide the reader with information for the objective and reproducible identification of pathologic MCs by using quantitative multiparametric flow cytometry, information for their phenotypic characterization, and the criteria currently used for a correct interpretation of the immunophenotypic results obtained.

show abstract

Immunophenotypic Analysis of Human Mast Cells by Flow Cytometry

Cited by 13 publications

References 24 publications

An immature immunophenotype of bone marrow mast cells predicts for multilineage D816V KIT mutation in systemic mastocytosis

An immature immunophenotype of bone marrow mast cells predicts for multilineage D816V KIT mutation in systemic mastocytosis

Evaluation of the WHO criteria for the classification of patients with mastocytosis

Immunophenotypic analysis of mast cells in mastocytosis: When and how to do it. Proposals of the Spanish Network on Mastocytosis (REMA)

Contact Info

Product

Resources

About