Rapid identification of monocytes in a mixed mononuclear cell preparation

Tucker, Stephen B.; Pierre, Robert V.; Jordon, Robert E.

doi:10.1016/0022-1759(77)90137-5

Cited by 304 publications

(90 citation statements)

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“…In a volume of 50 ml the cell suspension was added to Teflon culture bags and the cells were cultured for 10 days at 37" C in 5 % C02/95 % air. After 10 days in culture the macrophages were washed and again examined for nonspecific esterase activity and in some experiments for peroxidase activity (16). More than 95% of the macrophages were viable by a trypan blue exclusion test.…”

Section: Methodsmentioning

confidence: 99%

Phagocytosis-Associated Functions in Neonatal Monocyte-Derived Macrophages

et al. 1988

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A B S T R A n . Using a Teflon culture system we analyzed different aspects of cell maturation and phagocytic activities in neonatal monocyte-derived macrophages. Morphological and cytochemical characteristics as well as the protein composition of neonatal macrophages were identical with those of adult controls; actin binding protein (265,000 Da), myosin (210,000), a-actinin (102,000) and actin (42,000) could be identified in cells from either source: ~l l phagocytic functions were shown to be perfectly normal in neonatal macrophages when compared with adult cells: random migration, chemotactic response to zymosanactivated serum and formyl-methionyl-leucyl-phenylalanin, ingestion, and killing of Staphylococcus aureus, phagocytosis-associated chemiluminescence, production of oxygen intermediates (superoxide anion, 0;; hydrogenperoxide, H202). Phorbol myristate acetate-stimulated 0;-generation by 1 x lo5 macrophages was 11.8 f 4.7 nmol/h for neonates, and 10.2 f 3.9 for controls; production of H 2 0 2 was 7.6 f 3.5 nmol/h in neonatal macrophages and 6.4 +. In newborns, who suffer an increased susceptibility to serious systemic infections, subnormal phagocyte functions have been described. Neutrophils from neonates showed a decreased deformability (1) and a marked deficiency in movement toward different chemotactic stimuli (2-6). In addition generation of .OH by neonatal neutrophils was shown to be decreased (7). However, neonatal monocytes exhibited basically normal phagocytic functions when compared with adult cells (8). The functions of neonatal macrophages have not been fully explored. Macrophages from newborns synthesize humoral host defense factors such as complement components C2, C4 to a similar extent as phagocytes from adults (9, 10); fibronectin production Received October 12, 1987; accepted April 7, 1988. Correspondence Christian P. Speer, M.D., Universitats-Kinderklinik, RobertKoch-Str. 40, D-3400 Gottingen, Federal Republic of Germany.Supported by Grant Sp 239/2-2 from the Deutsche Forschungsgemeinschaft.2 1 was shown to be decreased in neonatal macrophages (9). Macrophages from newborns who had been activated by interferon-7 effectively killed or inhibited replication of toxoplasma gondii (1 I), presentation of bacterial Ag, and production of interleukin 1 was normal in cord blood monocytes/macrophages (12, 13). However, it has been suggested, that the decreased production of interferon-y by human neonatal cells (13) could be due to a functionally immature macrophage (14). Our study was initiated to examine the principal phagocyte activities of monocyte-derived macrophages in response to inflammation, i.e. chemotaxis, phagocytosis, killing, and generation of oxygen metabolites. Momholonical studies included a detailed analysis of the protein compos~ion of macrophages. MATERIALS AND METHODS Cell separation. Umbilical venous blood was withdrawn fromthe placenta into heparin (1-20 U/ml) immediately after delivery of healthy term newborns who were products of an uncomplicated pregnancy, normal labor, and...

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Section: Methodsmentioning

confidence: 99%

Phagocytosis-Associated Functions in Neonatal Monocyte-Derived Macrophages

et al. 1988

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“…Monocytes were purified from the mononuclear cell layer by adherence to plastic in RPMI-1640 medium with 10% FBS, I h, 37°C as described by Johnson et al (58). The monocytes were >70% pure as assessed by morphology and nonspecific esterase staining (59). Lymphocytes were purified from the nonadherent mononuclear cells using an additional overnight adherence step at 37°C in RPMI-1640 (Whittaker, M. A. Bioproducts) with 10% FBS; the nonadherent cells were >90% pure lymphocytes.…”

Section: Cell Types Evaluatedmentioning

confidence: 99%

Expression of the alpha-1-antitrypsin gene in mononuclear phagocytes of normal and alpha-1-antitrypsin-deficient individuals.

Mornex¹,

Chytil-Weir²,

Martinet³

et al. 1986

J. Clin. Invest.

166

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To evaluate the contribution of mononuclear phagocytes, and particularly alveolar macrophages, to alpha-l-antitrypsin (alAT) production in normal and alAT-deficient individuals, Northern analysis with a human alAT complementary DNA was used to demonstrate that alAT messenger RNA (mRNA) can be detected in liver, blood monocytes, and alveolar macrophages.Quantification of alAT mRNA expression demonstrated that: (a) type PiMM monocytes and alveolar macrophages expressed, respectively, 200-fold and 70-fold less alAT mRNA per cell than the liver, (b) the level of expression of the alAT gene was increased during the in vitro maturation of blood monocytes; and (c) blood monocyte and alveolar macrophage levels of expression of the alAT gene were the same in PiMM and PiZZ individuals. However, the amount of newly synthesized alAT secreted by ZZ alveolar macrophages was 10 times lower than that secreted by MM alveolar macrophages. Thus, mononuclear phagocytes of PiZZ individuals express a secretory defect in alAT in a fashion similar to hepatocytes. Not only do mononuclear phagocytes provide a readily accessible cell to evaluate the regulation of alAT gene expression, but these cells may contribute to the levels of alAT present in the lower respiratory tract in the nornul and ZZ states.

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“…They were designated as patients [21][22][23][24][25][26] to correspond with our ongoing series. Details on the other patients in the series can be found as follows: patients 1-9, [1][2][3][4][5][6][7]; patient 10, [8]; patients 11-16, [9]; patient 17, [16]; and patients 18-20, [17].…”

Section: Subjectsmentioning

confidence: 99%

“…Individual colonies were plucked from the cultures with a micropipette and spread on glass slides, and the cellular composition was determined microscopically using Wright's stain and nonspecific (a-naphthyl butyrate) esterase staining with and without treatment with fluoride [21]. Staining with benzidine was used to confirm intracellular hemoglobin in colonies scored as BFU-E.…”

Section: Clonogenic Assaysmentioning

confidence: 99%

Juvenile Chronic Myelogenous Leukemia Multilineage CD34⁺Cells: Aberrant Growth and Differentiation Properties

et al. 1996

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Rapid identification of monocytes in a mixed mononuclear cell preparation

Cited by 304 publications

References 2 publications

Phagocytosis-Associated Functions in Neonatal Monocyte-Derived Macrophages

Phagocytosis-Associated Functions in Neonatal Monocyte-Derived Macrophages

Expression of the alpha-1-antitrypsin gene in mononuclear phagocytes of normal and alpha-1-antitrypsin-deficient individuals.

Juvenile Chronic Myelogenous Leukemia Multilineage CD34⁺Cells: Aberrant Growth and Differentiation Properties

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Rapid identification of monocytes in a mixed mononuclear cell preparation

Cited by 304 publications

References 2 publications

Phagocytosis-Associated Functions in Neonatal Monocyte-Derived Macrophages

Phagocytosis-Associated Functions in Neonatal Monocyte-Derived Macrophages

Expression of the alpha-1-antitrypsin gene in mononuclear phagocytes of normal and alpha-1-antitrypsin-deficient individuals.

Juvenile Chronic Myelogenous Leukemia Multilineage CD34+Cells: Aberrant Growth and Differentiation Properties

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Juvenile Chronic Myelogenous Leukemia Multilineage CD34⁺Cells: Aberrant Growth and Differentiation Properties