Receptors for the Chemoattractants C5a and IL-8 Are Clustered on the Surface of Human Neutrophils
We have used high-resolution field emission scanning electron microscopy with backscatter electron imaging to detect immunogold-labeled C5a and interleukin-8 (IL-8) receptors on human blood neutrophils. The receptors were labeled with receptor-specific antibodies in combination with secondary antibo...
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Published in | The journal of histochemistry and cytochemistry Vol. 45; no. 11; pp. 1461 - 1467 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
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Los Angeles, CA
Histochemical Soc
01.11.1997
SAGE Publications |
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Abstract | We have used high-resolution field emission scanning electron microscopy with backscatter electron imaging to detect immunogold-labeled C5a and interleukin-8 (IL-8) receptors on human blood neutrophils. The receptors were labeled with receptor-specific antibodies in combination with secondary antibody conjugated to immunogold. When neutrophils were isolated in a “nonactivated” state, both of these receptor populations were expressed primarily in clusters on nonprojecting domains of the cell membrane. When these cells were double labeled for C5a and IL-8 receptors, intermixing of these receptor species in a common cluster was not found. When neutrophils were isolated in an “activated” state, by mixing the blood with N-formylmethionyl-leucyl-phenylalanine, the cells were seen to be elongated and ruffled at their anterior pole, but the C5a receptors did not disperse or redistribute on the surface of the peptide-activated cells. Analysis of the distribution of human C5a receptors expressed by transfected mouse L-cell fibroblasts showed the C5a receptors to be clustered, but expressed on nonprojecting and projecting domains of the cell surface. These observations provide new information on the topographical expression of leukocyte receptors involved in directing cell migration. (J Histochem Cytochem 45:1461–1467, 1997) |
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AbstractList | We have used high-resolution field emission scanning electron microscopy with backscatter electron imaging to detect immunogold-labeled C5a and interleukin-8 (IL-8) receptors on human blood neutrophils. The receptors were labeled with receptor-specific antibodies in combination with secondary antibody conjugated to immunogold. When neutrophils were isolated in a "nonactivated" state, both of these receptor populations were expressed primarily in clusters on nonprojecting domains of the cell membrane. When these cells were double labeled for C5a and IL-8 receptors, intermixing of these receptor species in a common cluster was not found. When neutrophils were isolated in an "activated" state, by mixing the blood with N-formylmethionyl-leucyl-phenylalanine, the cells were seen to be elongated and ruffled at their anterior pole, but the C5a receptors did not disperse or redistribute on the surface of the peptide-activated cells. Analysis of the distribution of human C5a receptors expressed by transfected mouse L-cell fibroblasts showed the C5a receptors to be clustered, but expressed on nonprojecting and projecting domains of the cell surface. These observations provide new information on the topographical expression of leukocyte receptors involved in directing cell migration. We have used high-resolution field emission scanning electron microscopy with backscatter electron imaging to detect immunogold-labeled C5a and interleukin-8 (IL-8) receptors on human blood neutrophils. The receptors were labeled with receptor-specific antibodies in combination with secondary antibody conjugated to immunogold. When neutrophils were isolated in a “nonactivated” state, both of these receptor populations were expressed primarily in clusters on nonprojecting domains of the cell membrane. When these cells were double labeled for C5a and IL-8 receptors, intermixing of these receptor species in a common cluster was not found. When neutrophils were isolated in an “activated” state, by mixing the blood with N-formylmethionyl-leucyl-phenylalanine, the cells were seen to be elongated and ruffled at their anterior pole, but the C5a receptors did not disperse or redistribute on the surface of the peptide-activated cells. Analysis of the distribution of human C5a receptors expressed by transfected mouse L-cell fibroblasts showed the C5a receptors to be clustered, but expressed on nonprojecting and projecting domains of the cell surface. These observations provide new information on the topographical expression of leukocyte receptors involved in directing cell migration. (J Histochem Cytochem 45:1461–1467, 1997) |
Author | Hasslen, Sharon R Carney, David F Gray, Gary D Herron, Michael J Nelson, Robert D Erlandsen, Stanley L Ember, Julia A |
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Cites_doi | 10.1016/0006-291X(91)91394-R 10.1111/j.1365-2818.1983.tb04170.x 10.1016/0167-5699(94)90314-X 10.1146/annurev.iy.12.040194.004015 10.1002/ar.1092410410 10.4049/jimmunol.151.1.377 10.1083/jcb.99.4.1461 10.1177/41.3.7679125 10.1021/bi00225a002 10.1073/pnas.85.15.5649 10.1177/44.8.8756756 10.1083/jcb.128.4.661 10.1177/44.10.8813076 10.4049/jimmunol.153.12.5682 10.1016/0092-8674(95)90278-3 10.1007/BF02388754 10.1016/0955-0674(94)90132-5 10.1016/0092-8674(95)90491-3 10.1016/0092-8674(91)90438-5 |
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Keywords | chemoattractant receptor chemokine IL-8 receptor immunocytochemistry field emission scanning electron microscopy (FESEM) mouse L-cells complement neutrophil C5a receptor (CD88) |
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SubjectTerms | Animals Antigens, CD - analysis Antigens, CD - genetics Female Fibroblasts - chemistry Humans Male Mice Microscopy, Electron, Scanning Microscopy, Immunoelectron Neutrophil Activation - immunology Neutrophils - chemistry Neutrophils - ultrastructure Receptor, Anaphylatoxin C5a Receptors, Complement - analysis Receptors, Complement - genetics Receptors, Interleukin - analysis Receptors, Interleukin-8A Transfection |
Title | Receptors for the Chemoattractants C5a and IL-8 Are Clustered on the Surface of Human Neutrophils |
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