In vivo labeling of resident peritoneal macrophages.
نویسندگان
چکیده
A novel method for labeling resident peritoneal macrophages (M phi) by injection of a dye into the peritoneal cavity is described. The dye, which fluoresces green, is selectively taken up by the resident M phi. Dye labeled cells can be further characterized by labeling of cell surface antigens with monoclonal antibodies (Mabs) and phycoerythrin conjugated second antibody. After such labeling with the Mabs F4/80 or Mac 1 the resident M phi were labeled by both the green dye and the red Mab markers, while recruited M phi 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 M phi in each animal without the use of radioisotopes, 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 M phi mediated cytotoxicity. We did find evidence that the i.p. injection induced 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 M phi, 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 M phi labeling occurred by a combination of phagocytosis of dye aggregates and endocytosis of labeled plasma membrane.
منابع مشابه
HETEROGENEITY OF MACROPHAGE POPULATIONS: ANTIBODIES DETECTING VARIOUS POPULATIONS OF MACROPHAGES
Two groups of antibodies against mouse resident alveolar and peritoneal macrophages have been raised individually in rabbits by immunization with whole cell suspensions of lavaged alveolar and peritoneal macrophages. The whole antisera reacted positively with neutrophils and lymphocytes. However, upon extensive absorptions with these cells the activity became restricted to macrophages. The...
متن کاملGlycolipids of the mouse peritoneal macrophage. Alterations in amount and surface exposure of specific glycolipid species occur in response to inflammation and tumoricidal activation
We have characterized the major glycolipid constituents of the mouse peritoneal macrophage, and have demonstrated that alterations in the amount and in the accessibility of specific glycolipid species to galactose oxidase/NaB3H4 labeling, an indicator of glycolipid surface exposure, occur in response to inflammation and as a consequence of activation to a tumoricidal state. The key findings are...
متن کاملCHROMOSOMAL PROTEINS IN PULMONARY ALVEOLAR MACROPHAGES
In this study the nature of chromosomal proteins, his tones and nonhistone in resident alveolar macrophages was investigated in comparison to peritoneal neutrophils and calf thymus proteins. Cells were obtained by lavaging the lung and after purity determination they were subjected to fractional extraction procedures. Proteins were then analysed on SDS polyacrylamide gels and densitometric...
متن کاملMacrophage origin limits functional plasticity in helminth-bacterial co-infection
Rapid reprogramming of the macrophage activation phenotype is considered important in the defense against consecutive infection with diverse infectious agents. However, in the setting of persistent, chronic infection the functional importance of macrophage-intrinsic adaptation to changing environments vs. recruitment of new macrophages remains unclear. Here we show that resident peritoneal macr...
متن کاملEffect of Peganum Harmala Seeds Extract on Nitric Oxide in U937 Monocytes and Macrophages
Background and Aims: Nitric oxide (NO) has an essential role in inflammation and has been related to pathogenesis and the progress of numerous inflammatory-based diseases, including some cancers. Peganum harmala (P. harmala) is a medicinal plant used for the treatment of numerous diseases such as several infections. Also, anti-inflammatory effects of P. harmala extracts and its derivatives (har...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Journal of leukocyte biology
دوره 43 5 شماره
صفحات -
تاریخ انتشار 1988