Identification of Basogranulin (BB1) as a Novel Immunohistochemical Marker of Basophils in Normal Bone Marrow and Patients With Myeloproliferative Disorders

Department of Internal Medicine I, Division of Hematology and Hemostaseology, Medical University of Vienna, Vienna, Austria.
American Journal of Clinical Pathology (Impact Factor: 3.01). 03/2006; 125(2):273-81. DOI: 10.1309/M9FQ-MQGF-6616-7N2X
Source: PubMed

ABSTRACT In myeloproliferative disorders (MPDs), basophils typically increase in number in the bone marrow (BM) and blood. In chronic myeloid leukemia (CML), basophilia is a diagnostic and prognostic variable. However, no reliable approach for routine detection and enumeration of basophils in BM sections is available. We applied the antibasogranulin antibody BB1 on paraffin-embedded BM sections in 21 control samples (normal BM), 45 patients with CML, 9 with chronic idiopathic myelofibrosis, 11 with polycythemia vera, 19 with essential thrombocythemia, and 7 with indolent systemic mastocytosis. As assessed by immunostaining of serial BM sections, BB1+ cells coexpressed myeloperoxidase, histidine decarboxylase, and leukosialin but did not express B- or T-cell-restricted antigens. BB1+ BM cells were found to be highly elevated in patients with CML compared with normal BM or other MPDs, with maximum counts found in accelerated phase CML (median, 160 cells/mm(2)). In summary, BB1 (basogranulin) is a new immunohistochemical basophil marker that should allow quantification of basophils in CML at diagnosis and during therapy.

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    • "In 16 patients with CP CML, 14 with AP, 6 with BP, and 5 with normal BM, expression of HGF was analyzed by IHC on serial sections (2 μm) prepared from formalin-fixed, paraffin-embedded, BM specimens. IHC was performed by the indirect immunoperoxidase staining technique as described [29] using a polyclonal anti-HGF antibody (work dilution, 1:50) and the anti-basophil mAb BB1 (1:300). After incubation (overnight) slides were washed, incubated with biotinylated second step goat anti-rabbit or horse anti-mouse antibodies (30 minutes), washed, and stained using 3-amino-9-ethyl-carbazole (Sigma-Aldrich). "
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    ABSTRACT: Chronic myeloid leukemia (CML) is a hematopoietic neoplasm characterized by the Philadelphia chromosome and the related BCR-ABL1 oncoprotein. Acceleration of CML is usually accompanied by basophilia. Several proangiogenic molecules have been implicated in disease acceleration, including the hepatocyte growth factor (HGF). However, little is known so far about the cellular distribution and function of HGF in CML. We here report that HGF is expressed abundantly in purified CML basophils and in the basophil-committed CML line KU812, whereas all other cell types examined expressed only trace amounts of HGF or no HGF. Interleukin 3, a major regulator of human basophils, was found to promote HGF expression in CML basophils. By contrast, BCR-ABL1 failed to induce HGF synthesis in CML cells, and imatinib failed to inhibit expression of HGF in these cells. Recombinant HGF as well as basophil-derived HGF induced endothelial cell migration in a scratch wound assay, and these effects of HGF were reverted by an anti-HGF antibody as well as by pharmacologic c-Met inhibitors. In addition, anti-HGF and c-Met inhibitors were found to suppress the spontaneous growth of KU812 cells, suggesting autocrine growth regulation. Together, HGF is a BCR-ABL1-independent angiogenic and autocrine growth regulator in CML. Basophils are a unique source of HGF in these patients and may play a more active role in disease-associated angiogenesis and disease progression than has so far been assumed. Our data also suggest that HGF and c-Met are potential therapeutic targets in CML.
    Neoplasia (New York, N.Y.) 07/2012; 14(7):572-84. DOI:10.1593/neo.12724 · 5.40 Impact Factor
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    • "For immunocytochemical (ICC) evaluation, primary MC were spun on cytospin slides and fixed in acetone. The ICC technique was performed as described previously (Agis et al., 2006) using a polyclonal rabbit-anti-human Kit antibody (Dako, "
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    ABSTRACT: Systemic mastocytosis (SM) in felines is a rare neoplasm defined by increased growth and accumulation of immature mast cells (MC) in various organs including the spleen. Although in many cases splenectomy is an effective approach, relapses may occur. In these patients, treatment options are limited. Recent data suggest that various Kit tyrosine kinase inhibitors (TKI) interfere with growth of neoplastic MC in humans. In the current study, we examined the effects of four TKI, imatinib, midostaurin, nilotinib, and dasatinib, on growth of spleen-derived feline neoplastic MC in three SM patients. Expression of Kit in neoplastic MC was confirmed by flow cytometry and/or Western blotting. In all three cases, a 12-bp internal tandem duplication in exon 8, resulting in a four amino acid-insertion between residues Thr418 and His419 in Kit, was detectable. As assessed by (3)H-thymidine incorporation experiments, all four TKI were found to inhibit the growth of feline neoplastic MC in a dose-dependent manner. The growth-inhibitory TKI effects were found to be associated with morphologic signs of apoptosis in MC. In conclusion, various Kit-targeting TKI can inhibit the in vitro growth and survival of feline neoplastic MC in SM.
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    • "Basophils normally are present in lower in numbers in normal bone marrow in comparison with peripheral blood and characteristically are increased in MPD, in particular chronic myeloid leukemia and myelofibrosis [23]. Occasionally, basophilia also is seen in association with MDS [24] or AML and infrequently in the absence of neoplasia. "
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    ABSTRACT: As hematopoietic cells proceed in differentiation from stem cells to committed progenitors to later stage mature forms, they undergo a sequence of morphologic, immunophenotypic, and functional changes that are a consequence of interaction between the underlying cellular genetic program and environmental cues, are linear for each cell lineage, and result in a pattern of antigenic expression related to lineage and stage of maturation. The antigenic patterns characteristic of normal maturation have been elucidated systematically and found invariant between individuals. Deviation from this pattern is a hallmark of hematopoietic neoplasia. Application of these principles to myelodysplastic syndromes, myeloproliferative disorders, and acute myeloid leukemia is presented and illustrated.
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