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Journal of Leukocyte Biology 43:387-397 (1988) © 1988 Alan R. Liss, Inc. In Vivo Labeling of Resident Peritoneal Macrophages
BibTeX
@MISC{Melnicoff_journalof,
author = {Meryle J. Melnicoff and Bruce D. Jensen and Elizabeth W. Breslin and Paul K. Horan and Department Of Immunology},
title = {Journal of Leukocyte Biology 43:387-397 (1988) © 1988 Alan R. Liss, Inc. In Vivo Labeling of Resident Peritoneal Macrophages},
year = {}
}
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Abstract
A novel method for labeling resident peritoneal macrophages (MO) by injection of a dye into the peritoneal cavity is described. The dye, which fluoresces green, is selectively taken up by the resident MO. Dye labeled cells can be further characterized by labeling of cell surface antigens with monoclonal antibodies (Mabs) and phycoerythrln conju-gated second antibody. After such labeling with the Mabs F4180 or Mac 1 the resident MQ were labeled by both the green dye and the red Mab markers, while recruited MQ or neutrophils were labeled with just the red Mab; the two populations of cells were readily distinguished by two.color flow cytometry. This technique enabled identification of resident and recruited MO in each animal without the use of radloisotopes, irradiation, or bone marrow ablation. Sufficient numbers of cells can be analyzed from each animal so that individual animals could be evaluated. We found no adverse effects of this labeling technique on expression of cell surface antigens or MO mediated cytotoxicity. We did find evidence that the i.p. injection in-duced a mild inflammation in the peritoneal cavities of animals injected with either the dye or the balanced salt solution vehicle. Examination of the intracellular staining pattern indicated that the label rapidly sequestered in the cytoplasm of the MO, possibly in the lysosomes. Dye solubility studies showed that the dye was partially soluble at the concentration used for in vivo labeling. We hypothesize that the MQ labeling occurred by a combination of phagocytosis of dye aggregates and endocytosis of labeled plasma membrane.
Keyphrases
resident peritoneal macrophage vivo labeling leukocyte biology cell surface peritoneal cavity resident mo intracellular staining pattern labeling technique resident mq balanced salt solution vehicle labeled plasma membrane bone marrow ablation mq labeling novel method monoclonal antibody green dye phycoerythrln conju-gated second antibody dye solubility study individual animal color flow cytometry mabs f4180 red mab adverse effect sufficient number red mab marker mild inflammation dye aggregate
