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  • 1
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background Mast cells (MCs) arise from haematopoietic stem cells. We have recently reported that CD34+ progenitors derived from human bone marrow (BM) develop into tryptase+, chymase+ MCs when cultured in the presence of recombinant human stem cell factor (rhSCF) and recombinant human IL-6 (rhIL-6). In an experiment for the expression of chymase during differentiation, chymase+ cells were detected in human BM, but tryptase+ cells were not found.Objective The purpose of this study was to show the appearance of chymase+ cells in CD34+ cells with an origin different from MC differentiation.Methods CD34+ cells from human BM were sorted with anti-CD117 monoclonal antibody (mAb), and cytospins of CD34+, CD34+CD117+, or CD34+CD117− were prepared. These cells were cultured with rhSCF+rhIL-6 for 12 weeks. Some of the cells were subjected to either histological stain with Wright–Giemsa or immunocytochemistry with anti-chymase mAb. Real-time RT-PCR was also performed to compare the transcriptional level of chymase from each cell preparation.Results Chymase was expressed in CD34+ cells as well as human MCs by immunocytochemistry. Substantial CD34+CD117− cells, but not CD34+CD117+ cells, were stained immunocytochemically with anti-chymase mAb. For 1 week culture with rhSCF+rhIL-6, no cells expressed chymase in any preparation. Real-time RT-PCR revealed positivity for chymase mRNA in CD34+ cells, but it reduced at 1 week of culture, and increased as cells developed into MCs. Chymase mRNA in CD34+CD117+ cells was negligible compared with that in CD34+CD117−. Tryptase mRNA was below the detectable level in CD34+ cells, and increased along with MC differentiation. After 12 weeks of culture, CD34+CD117+ developed predominantly into MCs, whereas CD34+CD117− developed into monocytes/macrophages.Conclusion Our findings suggested that chymase is present not only in MCs but also in CD34+CD117− BM progenitors, but that its origin is different from the MC lineage.
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  • 2
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background In vitro-derived human mast cells exhibit different properties, depending in part on the source of progenitor cells. Most investigations have used fetal liver, cord blood or peripheral blood. Few have used adult bone marrow.Objective Human mast cells derived in vitro from the CD34+ progenitors in bone marrow and cord blood that had been cultured with recombinant human stem cell factor (rhSCF) and recombinant human interleukin-6 (rhIL-6) were compared.Methods and results After 12 weeks of culture, nearly all of the cells were mast cells, and nearly all of these had cytoplasmic granules containing both tryptase and chymase (MCTC type), stained metachromatically with acidic toluidine blue, and expressed CD117 on the cell surface. Both tryptase protein and mRNA were detected by two weeks of culture. Chymase mRNA and protein were detected at 4 weeks but not at 2 weeks of culture. By 12 weeks, chymase content per cell, measured by ELISA, was significantly higher (P 〈 0.05) in human bone marrow-derived mast cells (HBMMC) (5.6 ± 0.9 pg) than in cord blood-derived mast cells (CBMC) (2.4 ± 0.9 pg), whereas histamine and tryptase levels were not significantly different. Of the cluster designations tested, CD29, CD49d, CD51 and CD61 were strongly expressed on HBMMC. CD54 and FcεRIα also were expressed constitutively. Approximately half of CD34-sorted cells at day 0 were CD13+ and this diminished as mast cell maturation occurred. Electron microscopy revealed that 12-week-old HBMMC had many secretory granules that contained spherical electron dense cores surrounded by electron lucent space, consistent with previous reports of immature MCTC cells developing in vivo.Conclusions CD34+ progenitors of human bone marrow are a rich source of mast cell progenitors capable of expressing granule and surface markers of mature mast cells in the presence of rhSCF and rhIL-6.
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  • 3
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background CD34+ progenitor cells develop into tryptase+, CD117+ mast cells when cultured in the presence of recombinant human stem cell factor (rhSCF). However, spontaneous IgE receptor (FcɛRI) expression during human mast cell development is not well examined.Objective Here, the expression and function of FcɛRI in and on human bone marrow-derived mast cells (HBMMCs) during development were investigated.Methods and Results At 4 weeks of culture, predominant cells expressed high-affinity IgE receptor α chain (FcɛRIα) on the cell surface determined by flow cytometry, but CD117 was less expressed. Immunocytochemistry with antitryptase mAb and anti-FcɛRIα mAb revealed intracellular and surface expression of FcɛRIα at 2 weeks of culture, but tryptase was less expressed. FcɛRIα mRNA transcript preceded that of tryptase mRNA at 2 weeks of culture determined by real-time RT-PCR, and FcɛRIα, FcɛRIβ, FcɛRIγ, and tryptase mRNA increased along with differentiation. FcɛRIα cross-link on HBMMC and 4-week-old mast cells/mast cell precursors induced the release of IL-5 and granulocyte macrophage-colony stimulating factor, which was enhanced by rhSCF.Conclusion These data indicated that HBMMC constitutively and spontaneously expressed functional FcɛRI subunits at the early stage of differentiation, probably because of the differences in the ability and functional property of progenitors.
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  • 4
    ISSN: 1437-7772
    Keywords: Key words Humoral hypercalcemia of malignancy (HHM) ; Lung cancer ; Psoas abscess-like metastasis ; Parathyroid hormone-related protein (PTH-rP)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We report a patient with lung cancer associated with psoas abscess-like metastasis and humoral hypercalcemia of malignancy (HHM). A 71-year-old man was hospitalized with exacerbating pain on the left-side of his back and thigh. A computed tomography (CT) scan of the abdomen demonstrated a tumor in the left iliopsoas muscle, first diagnosed as psoas abscess. A chest roentgenogram showed right-sided pleural effusion, and CT scan of the chest demonstrated a tumor in the right lower lobe of the lung. His serum calcium level was 12.8 mg/dl, and parathyroid hormone-related protein (PTH-rP) level was elevated, to 11.1 pmol/l. The patient died of progressive disease, and necropsy specimens were diagnosed as poorly differentiated squamous cell carcinoma. Immunohistochemical staining for PTH-rP was positive in the tumor cells, including the primary and the metastatic lesions. A review of this unusual tumor type is also presented.
    Type of Medium: Electronic Resource
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