Different functional and morphological characteristics in a nonadherent subpopulation of human macrophages recovered by bronchoalveolar lavage

Sköld, C. Magnus; Barck, C.; Lundahl, Joachim; Johansson, Anne

doi:10.1183/09031936.95.08101719

Cited by 12 publications

(7 citation statements)

References 16 publications

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“…AM recovered from adult and senescent animals showed a higher percentage of dark blue reactive cells (colour intensity is associated with the cells' ability to produce ROS) after stimulation with TPA or LPS + TPA when compared with AM from young animals. This finding may be due partly to the heterogeneity in the adhesive properties and phagocytic capacities found in AM recovered by BAL from human healthy smokers as reported by Skold et al . (1995).…”

Section: Discussionmentioning

confidence: 56%

Age‐dependent change in reactive oxygen species and nitric oxide generation by rat alveolar macrophages*

et al. 2003

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SummaryImmunosenescence is an age-associated dysregulation of the immune function, which contributes to increased susceptibility to disease in the elderly. Alveolar macrophages (AM) are known phagocytes that generate reactive oxygen species (ROS) and nitric oxide (NO), essential mediators for host defence. We studied phagocytosis, ROS and NO production in AM obtained from young, adult and senescent rats (1-2, 9-12 and 18-24 months old, respectively) after exposure to lipopolysaccharide (LPS, 0.1-10 µ µ µ µ g mL), 12-O-tetradecanoylphorbol 13-acetate (TPA, 0.1 µ µ µ µ g mL − − − − 1 ) or LPS + TPA in culture. Phagocytosis was significantly lower in control AM from adult rats than in AM from young animals. Nevertheless, AM from adult animals pretreated with LPS exhibited higher phagocytic capacity than AM from younger animals. ROS was identified by the NBT test at single cell level and quantified by automated image analysis. When TPA was added to all three populations, AM from adult and senescent animals responded more than AM from young animals. All LPS-stimulated AM produce more NO than controls. However, NO production increased three-, four-and two-fold in young, adult and senescent animals, respectively. Our results demonstrate that AM from young, adult and senescent animals display differential responsiveness to inflammatory mediators. Therefore, aging processes markedly affect AM metabolic functions and may further compromise the lung immune defence response, increasing adverse long-term health effects.

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Section: Discussionmentioning

confidence: 56%

Age‐dependent change in reactive oxygen species and nitric oxide generation by rat alveolar macrophages*

et al. 2003

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“…Their microscopic appearance at the time of sorting was similar to that reported for alveolar macrophages found within bronchoalveolar lavage samples (7, 12). Interestingly, intense autofluorescence among alveolar macrophages has also been associated with a lack of adherence in culture assays (22). Our cells also share morphological similarities with so‐called “foam” cells obtained directly from atherosclerotic lesions (23).…”

Section: Discussionmentioning

confidence: 99%

Characterization of viable autofluorescent macrophages among cultured peripheral blood mononuclear cells

et al. 2001

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Background Inflammatory macrophages that demonstrate intense autofluorescence have been isolated directly from alveolar and peritoneal tissues, but their generation in vitro remains vague. We use flow cytometry to identify a population of autofluorescent macrophages as they arise among nonadherent populations of cultured blood mononuclear cells. Methods Cells were obtained from donated blood buffy coats and placed in culture for 14 days. Unstained populations from the cells remaining in suspension were sampled daily using flow cytometry. During the first 5 culture days, a distinct population of autofluorescent cells arose and comprised an average of ≤14% of the total cell population. This population declined to less than 6% by culture day 8. Results The cells were identified as viable macrophages expressing CD68, lysozyme, and HLA‐DR. Quantitative reverse transcription‐polymerase chain reaction (RT‐PCR) demonstrated a unique cytokine profile with IL‐1α expression levels 138‐fold higher than those measured in uncultured monocytes. No significant elevation in the levels of other cytokines was identified. Upon replating, the sorted populations became readherent, were able to ingest plastic beads, and remained viable for 6 or more additional weeks in culture without evidence of proliferation or multinucleation. Conclusion Viable autofluorescent macrophage populations arising among cultured peripheral blood may be easily identified and isolated for further study using flow cytometry. Cytometry 44:38–44, 2001. Published 2001 Wiley‐Liss, Inc.

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“…Adhesion assay. The adhesion assay was performed as described previously [16]. Briefly, culture wells, with a growth area of 3.83cm 2 (art.…”

Section: Methodsmentioning

confidence: 99%

Monocyte and Neutrophil Adhesion to Matrix Proteins is Selectively Enhanced in the Presence of Inflammatory Mediators

et al. 1996

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The authors investigated the time course of monocyte and neutrophil adhesion to fibronectin, vitronectin and albumin precoated culture wells, using mixed leucocyte populations from healthy blood donors. Moreover, the influence of chemotactic agonists on the adhesion properties as well as the quantitative expression of CD29, CD11b/CD18 and CD61 was analysed by flow cytometry. Different chemotactic agonists were used representing a classical chemotactic agonist (fMLP), and agonists with a preferential effect on monocytes (RANTES) and neutrophils (IL-8), respectively. The authors found a gradual increase in monocyte and neutrophil adhesion to all three surfaces, reaching a plateau at 15 min of incubation. Adhesion to fibronectin was significantly higher at all time points (5, 15 and 60 min, respectively) compared with vitronectin and albumin in both monocytes and neutrophils. Neutrophil adhesion to vitronectin was significantly lower at 60 min compared with 15 min. Monocyte adhesion to albumin was increased by fMLP and RANTES and to vitronectin also by IL-8. Neutrophil adhesion to albumin and vitronectin was increased by fMLP and IL-8, but not RANTES. The adhesion to fibronectin was not altered by any of the chemotactic agonists used. The quantitative levels of CD11b/CD18, but not CD29 and CD61, was increased by fMLP, but not RANTES nor IL-8. The authors conclude that the adhesion of human monocytes and neutrophils to vitronectin and albumin, but not fibronectin, is selectively enhanced by chemotactic agonists and may contribute to the selective accumulation of different leucocyte subsets at the inflammatory site.

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Different functional and morphological characteristics in a nonadherent subpopulation of human macrophages recovered by bronchoalveolar lavage

Cited by 12 publications

References 16 publications

Age‐dependent change in reactive oxygen species and nitric oxide generation by rat alveolar macrophages*

Age‐dependent change in reactive oxygen species and nitric oxide generation by rat alveolar macrophages*

Characterization of viable autofluorescent macrophages among cultured peripheral blood mononuclear cells

Monocyte and Neutrophil Adhesion to Matrix Proteins is Selectively Enhanced in the Presence of Inflammatory Mediators

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