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: 2.51). 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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Available from: Andrew F Walls, Sep 29, 2015
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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 · 4.25 Impact Factor
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    • "So far, only very few immunohistochemical markers sufficient for the evaluation of basophils and eosinophils in BM sections are available. For basophil evaluation and counting, 2D7 and BB1 (basogranulin) are recommended antigens (Figure 1E) [23,24] while eosinophils can be visualized using an antibody against eosinophil major basic protein (EMBP) (Figure 1F). Whereas a slight or moderate increase of eosinophils is often seen in reactive and neoplastic disease states, constant basophilia is uncommon in reactive states and thus regarded as a potential indicator for the presence of a myeloid neoplasm. "
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    ABSTRACT: The diagnosis, classification, and prognostication of patients with myelodysplastic syndromes (MDS) are usually based on clinical parameters, analysis of peripheral blood and bone marrow smears, and cytogenetic determinants. However, a thorough histologic and immunohistochemical examination of the bone marrow is often required for a final diagnosis and exact classification in these patients. Notably, histology and immunohistology may reveal dysplasia in megakaryocytes or other bone marrow lineages and/or the presence of clusters of CD34-positive precursor cells. In other cases, histology may reveal an unrelated or co-existing hematopoietic neoplasm, or may support the conclusion the patient is suffering from acute myeloid leukemia rather than MDS. Moreover, histologic investigations and immunohistology may reveal an increase in tryptase-positive cells, a coexisting systemic mastocytosis, or bone marrow fibrosis, which is of prognostic significance. To discuss diagnostic algorithms, terminologies, parameters, and specific issues in the hematopathologic evaluation of MDS, a Working Conference involving a consortium of US and EU experts, was organized in June 2010. The outcomes of the conference and resulting recommendations provided by the faculty, are reported in this article. These guidelines should assist in the diagnosis, classification, and prognostication in MDS in daily practice as well as in clinical trials.
    Oncotarget 11/2010; 1(7):483-96. DOI:10.18632/oncotarget.185 · 6.36 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.
    Veterinary Immunology and Immunopathology 06/2009; 132(2-4):243-50. DOI:10.1016/j.vetimm.2009.05.007 · 1.54 Impact Factor
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