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1.
Summary Using ferritin as a marker of reactive microglia, we demonstrated a close association between proliferation of reactive microglia and expression of human immunodeficiency virus type 1 (HIV-1) in brain tissue from autopsied cases of acquired immunodeficiency syndrome (AIDS). An increased number of ferritin-positive reactive microglia was observed in formalin-fixed paraffin-embedded brain sections from all 13 AIDS cases examined. Similar findings were observed in brain tissue from other neurological diseases (subacute sclerosing penencephalitis, herpes simplex encephalitis and multiple sclerosis). Multinucleated giant cells were found in 7 of the AIDS cases which were also intensely labeled for ferritin. Dual-label immunohistochemistry using anti-ferritin and cell-specific markers showed that ferritin-positive cells were distinct from astrocytes, neurons and endothelia using anti-glial fibrillary acidic protein (anti-GFAP), anti-neurofilament protein and Ulex europaeus agglutinin 1, respectively. In 5 AIDS brains, only ferritin-positive cells were shown to contain HIV-1 gp41 antigen using dual-label immunohistochemistry. In addition, HIV-1 RNA was localized in territin-positive reactive microglia but not in GFAP-positive astrocytes using immunohistochemistry combined with in situ hybridization. Ferritin-positive reactive microglia and multinucleated giant cells were colabeled with the microglial marker, Ricinus communis agglutinin 1 (RCA-1). Howerver, RCA-1 also extensively stained resting microglia only a few of which were colabeled for ferritin. The density of ferritin-positive cells was correlated with the presence of HIV-1 RNA-positive cells in AIDS brain. Thus, ferritin immunoreactivity can be used as an activation marker of microglia in archival paraffin sections and reflects the extent of inflammation in HIV-1-infected brain.Supported in part by NIH Grants RO1 DA04787, RO1 HD26621, PO1 NS25569, the Biopsychosocial Center for the Study of AIDS (NIMH P50 MH 43455), the Department of Veterans Affairs, the Mary Jane Crowe Foundation, the Swedish Society of Medicine (Stockholm, Sweden), and the Multiple Sclerosis Society of Göteborg (Göteborg, Sweden)  相似文献   

2.
The lesions in periventricular leukomalacia (PVL) comprise necrosis and a glial reaction in the deep cerebral white matter of fetuses and neonates. The purpose of this study was to elucidate the role of glial cells in the formation of the lesions in PVL. Ten PVL brains and 22 control brains were immunohistochemically compared using anti-glial fibrillary acidic protein (GFAP) and anti-ferritin antibodies, and lectin-Ricinus communis agglutinin (RCA-1). The numbers of GFAP-positive glia and RCA-1 positive glia increased in the whole white matter and the periventricular white matter, respectively. Ferritin was predominantly stained in the cytoplasm of oligodendrocytes, and the number of ferritin-positive oligodendrocytes gradually increased with age in normal brains. However, ferritin was stained in microglia and partially reactive astrocytes, instead of ferritin-positive oligodendrocytes, in PVL brains. A relationship of the glial cellular reaction with the time scale of histologic change in PVL was shown by the appearance of RCA-1-positive microglia around fresh necrotic regions, accumulation of RCA-1-positive macrophages in necrotic regions, and then proliferation of GFAP-positive reactive astrocytes with long processes outside the microglial reaction sites. On cavity formation, the end-stage of PVL, rough walls of moderately dense gliosis consisted of slightly GFAP-positive fibrillary astrocytes. PVL involves glial cellular reactions not only in regional necrotic lesions, but also in the whole cerebral white matter. The decrease in ferritin-positive oligodendrocytes in PVL brains may be related to the delayed myelination in the brains of long-surviving infants with PVL.  相似文献   

3.
We have used an immunocytochemical approach to investigate the inter-relationships between astrocytes, macrophages and microglia and the extracellular matrix components fibronectin and laminin, in 27 gliomas. Using recently available markers to macrophages and microglia (monoclonals Mac387, KP1 and the lectin RCA-1) and antisera to GFAP, fibronectin and laminin, we have described the reactions of these cells and the extracellular matrix. We found RCA-1 to be the superior marker for detecting most macrophages and microglia. There were more macrophages and microglia in high-grade gliomas than in low-grade. RCA-1 also reacted with endothelial cells in normal and reactive brain but showed markedly less affinity for endothelium in an close to gliomas. A possible role for the extracellular matrix in the astrocyte, macrophage and microglial reactions is discussed in the broader context of their role in the immune response.  相似文献   

4.
For comparison with an earlier paper on gliomas (Morris and Esiri, J. Neurol. Sci., 101 (1991) 47-58), we have used an immunocytochemical approach to examine the reactions of astrocytes, macrophages and microglia and alterations to the extracellular matrix components fibonectin and laminin in 15 cases of non-neoplastic CNS disease. We compared recently available markers for the detection of macrophages and microglia (monoclonals Mac387, KP1 and the lectin RCA-1) and also used antisera to GFAP, fibronectin and laminin. RCA-1 was the superior marker for detecting macrophages and microglia but it also reacted with endothelial cells in normal and reactive brain. The numbers of macrophages and microglia were highly variable, depending on the type of lesion. Fibronectin and laminin were largely confined to the vasculature and leptomeninges. The relationship between the extracellular matrix and the astrocyte, macrophage and microglial reactions is discussed.  相似文献   

5.
In neuropathological studies it is important to detect both resting and reactive microglia in paraffin sections. We examined the usefulness of human (h) GLUT5, a glucose transporter, as a microglial marker. We produced an hGLUT5 antibody against its C-terminal sequence and stained human brain tissue sections. The hGLUT5 antibody consistently stained microglia in cryostat sections. In paraffin sections fixed with formalin, paraformaldehyde or ethanol, both resting and reactive microglia were stained; the latter were stained more intensely than the former. The hGLUT5 and glial fibrillary acidic protein labeling did not overlap each other in double immunofluorescence analyses. Oligodendrocytes, perivascular cells, choroid plexus epithelium and ependymal cell were negative for hGLUT5. Even after 1-month fixation in formalin, the hGLUT5 antibody stained microglia well. Microwave pretreatment enhanced the immunoreactivity of hGLUT5. As compared with other microglial markers, KP-1, KiM1p, CR3.43 and RCA-1, the hGLUT5 antibody could be considered good morphological marker. hGLUT5 immunolabeling clearly showed the detailed microglial processes, whereas immunolabeling with Ki-M1P and KP-1 showed cytoplasmic granules, and it was difficult to trace the microglial processes. The hGLUT5 antibody stained both resting and reactive microglia, whereas CR3.43 stained only reactive microglia, and RCA-1 labeled microvessels more intensely than microglia. Thus, hGLUT5 is a marker that is suitable for routine histopathological staining procedures.  相似文献   

6.
Brain hemosiderin and superficial siderosis of the central nervous system   总被引:7,自引:0,他引:7  
Brain tissue from five patients with superficial siderosis of the central nervous system was examined by immunocytochemistry for ferritin, glial fibrillary acidic protein (GFAP), alpha 1-antitrypsin, and alpha 1-antichymotrypsin, and by lectin affinity cytochemistry with biotinylated Ricinus communis agglutinin-1 (RCA-1). The sections were pretreated with 2,2'-dipyridyl and sodium hydrosulfite to remove iron and to reveal the antigenic sites. In siderotic cerebellar cortex, ferritin reaction product occurred in the hemosiderin matrix, the cell bodies and processes of Bergmann glia, and in microglia. Astrocytes other than Bergmann glia did not contain ferritin reaction product. RCA-1 stained microglia and hemosiderin whereas antisera to alpha 1-antitrypsin and alpha 1-antichymotrypsin only reacted with iron-depleted granules. The selective vulnerability of the eighth cranial nerve was explained by the presence of ferritin-reactive and lectin-positive microglia. Hemosiderin isolated from frozen cerebellum contained ferritin, GFAP, and vimentin. The presence of the intermediate filament proteins was likely due to co-localization with hemosiderin granules in Bergmann glia. The ability of the brain to biosynthesize ferritin in response to prolonged contact with hemoglobin iron is thought to be the most important factor in the pathogenesis of superficial siderosis. The great severity of the lesion in the exposed cerebellar cortex is readily explained by accelerated ferritin biosynthesis in Bergmann glia.  相似文献   

7.
Mistletoe lectin-1 (ML-1) and Ricinus communis agglutinin-120 (RCA-1) both possess D-galactose-specific surface-binding sites. They were used to selectively identify microglial populations in aldehyde-fixed normal brain tissue by lectin immunohistochemistry on paraffin and frozen sections. Mistletoe lectin-1 was superior to RCA-1 in labelling microglia in the rat brain, whereas RCA-1 labelled human microglia better than ML-1. Thus, RCA-1 and ML-1 supplement each other for identifying microglial in human and rodent central nervous system tissues. The high reproducibility of the results and the applicability of the technique to routine histology, using formalin-fixed tissue, should facilitate study of the histogenesis and role of microglia in the CNS.  相似文献   

8.
Differential morphologic subtypes of microglia have been identified in the human fetal frontal cerebrum using a lectin, Ricinus communis agglutinin 1 (RCA-1), and a monoclonal antibody, EBM-11. In this report, microglia were characterized in the human fetal cervical spinal cord. RCA-1-positive microglia were ramified in the developing gray matter while in the developing white matter they had a less differentiated (ameboid) appearance. EBM-11, a monoclonal antibody that recognizes CD68 on human macrophages, and microglia labeled only ameboid-type microglia in the developing white matter. This suggests that distinct subpopulations of microglia exist, which may represent different stages in microglial development, and that CD68 may be a differentiation marker for less mature forms. Therefore, cytologically less differentiated forms of microglia appear to be associated with myelination.  相似文献   

9.
Our previous studies indicate that glucose transporter 5 (GLUT5) is a microglial marker in routine paraffin sections, and is rarely present in monocytes/macrophages of the peripheral organs. We examined the expression of GLUT5 in 91 cases of human gliomas to characterize the microglial phenotype in glioma tissues. Immunohistochemistry was performed on formalin-fixed, paraffin-embedded sections using such antibodies as a GLUT5 antibody, two markers for activated microglia: major histocompatibility complex (MHC) class II Ag and macrophage scavenger receptor class A (MSR-A), and MIB-1 antibody. The immunoreactivity of GLUT5 was present in three microglial phenotypes: ramified (resting), activated, and ameboid (macrophagic) microglia in most of the cases. A double-labelling study of astrocytic tumours using GLUT5 and MIB-1 antibodies demonstrated a proportion of proliferating microglia. However, no morphological difference between MIB-1-positive, microglial cells and MIB-1-negative, microglial cells was found. The number of GLUT5-positive microglia was significantly (P < 0.001) higher in astrocytic tumours than in oligodendroglial tumours. Many GLUT5-positive microglia (up to 52% in total cells) were often observed in pilocytic astrocytomas, where microglial cells were predominantly ramified, and the number of MHC class II- or MSR-A-positive microglia was less than GLUT5-positive microglia. Thus, the present study indicated that intrinsic microglia can be a source of microglia/macrophages cell populations in astrocytic tumours, and that pilocytic astrocytomas often have a high proportion of microglial cells with mild activation.  相似文献   

10.
A new immunoperoxidase marker for microglia in paraffin section   总被引:2,自引:0,他引:2  
We tested antisera against monocytic and lymphocytic lineages on 27 formalin-fixed, paraffin-embedded brains, obtained at autopsy, using the avidin-biotin-complex immunoperoxidase method. Patients ranged from one day to 69 years with 12 cases under the age of two. LN-1-positive microglia were present in 22 brains. LN-1 did not stain any other glial or neuronal cells. Five negative brains included two irradiated gliomas, two cases of multiple sclerosis and one normal case. Immunostaining was confined to cells with bipolar processes and rod-shaped nuclei recapitulating the characteristic features of microglia in silver-impregnated sections. LN-1-positive microglia were most prominent in the grey matter and in the grey-white junction with fewer positive cells seen in the white matter. Double immunostaining with LN-1 and glial fibrillary acidic protein (GFAP) clearly distinguished LN-1-positive microglia and GFAP-positive astrocytes. The expression of LN-1, a B-lymphocyte antigen, by microglia contradicts the macrophage derivation theory and supports data indicating a functional role of microglia in immune processes. LN-1, while not specific for microglia, should be considered a useful marker, more reliable than silver impregnation, for detecting microglia in paraffin section.  相似文献   

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