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  • 1
    ISSN: 1432-0533
    Keywords: Key words Cell culture ; Cell line ; Glioma ; Calcium-binding proteins ; Microglia enzymology ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Allograft inflammatory factor-1 (AIF-1) is a Ca2+-binding peptide that constitutes a potential modulator of macrophage activation and function during the immune response of the brain. Peptides termed microglia response factor-1 or ionized calcium-binding adaptor molecule-1 have been reported to be identical with AIF-1. We have investigated the expression of AIF-1 in the rat C6 glioblastoma and 9L gliosarcoma tumor models and additionally assessed AIF-1 expression in a diverse range of human astrocytomas by immunohistochemistry. AIF-1 was expressed by activated microglial cells and a subset of infiltrating macrophages in areas of infiltrative tumor growth and in compact tumor areas in both rat and human gliomas. Double-labeling experiments in rats and humans characterized the nature and the functional status of AIF-1+ cells. AIF-1 expression was detected in cells expressing major histocompatibility complex class II molecules and in a subset of activated macrophages/microglial cells. All MRP-8+ cells coexpressed AIF-1. In humans, there was a strong correlation of AIF-1-expressing activated macrophages/microglial cells with tumor malignancy (P 〈 0.0001). These results suggest that AIF-1 defines a distinct subset of tumor-associated activated macrophages/ microglial cells.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Histochemistry and cell biology 110 (1998), S. 425-430 
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Here we demonstrate a simple and reliable multiple epitope labeling technique based exclusively on the alkaline phosphatase (AP) enzyme-linked visualization method. AP is functionally blocked by ethylenediaminetetraacetic acid (EDTA), which allows the repeated use of AP conjugates in immunohistochemistry with different substrates. We found that reactivation of AP function following EDTA incubation is dependent on EDTA concentration and incubation time. While incubation times of up to 2 h in 0.25 M EDTA, pH 6, exhibit a resumption of AP enzyme function, more than 2 h of incubation irreversibly blocks AP enzyme activity. Surprisingly, EDTA incubation also results in considerable but not complete inhibition of antibody crossreactivity during immunohistochemistry. Thus, this technique is suitable for single-layer, multiple-staining experiments with AP-linked primary antibodies or multilayer labeling with antibodies of various species for sequential staining rounds. We demonstrate the applicability of this technique in immunohistochemistry by double-labeling experiments using the monoclonal antibodies anti-glial fibrillary acidic protein, anti-leucocyte common antigen, anti-CD43/CD45RA (pan-human leucocyte), and anti-migration inhibitory factor-related protein-8 in combination with an in situ nick translation assay to characterize differentiating antigens of apoptotic cells in human glioblastoma paraffin sections.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0533
    Keywords: Key words Glioma ; Cyclooxygenase ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Cyclooxygenases (COX, prostaglandin endoperoxide synthases, PGG/H synthases) are potent mediators of inflammation. While COX-1 is constitutively expressed in a wide range of tissues, COX-2 is cytokine inducible. Although COX-1 expression is observed in normal tissue, enhanced COX-2 expression has been attributed a key role in the development of edema, impeding blood flow and immunomodulation observed in pathologically altered tissues. Here, we have analyzed the expression of COX-1 and COX-2 in 50 gliomas and 10 control brains with no neuropathological alterations by immunohistochemistry; 22 glioblastoma multiforme, 9 anaplastic astrocytomas, 5 protoplasmic astrocytomas, 1 gemistocytic astrocytoma and 13 fibrillary astrocytomas were included in the study. Compared with control brains, accumulation of COX-1 was detected in 20–50% of all cells in both low- and high-grade gliomas. Double-labeling experiments revealed COX-1 expression in subsets of macrophages/ microglial cells within the tumor parenchyma and in areas of infiltrative tumor growth. Of the COX-1-positive cells, 90% expressed MHC class II antigens. No COX-1 immunoreactivity was observed in tumor cells. COX-2-positive cells accumulated in tumor cells and in single macrophages/microglial cells in the immediate vicinity of necroses. Further studies are required to determine whether COX-2 is involved in the development of necrosis or, more likely, whether COX-2 is a part of the tumor tissue response to necrosis.
    Type of Medium: Electronic Resource
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