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Phagocytosis in Mesocestoides vogae-induced peritoneal monocytes/macrophages via opsonin-dependent or independent pathways

G. Hrčková
G. Hrčková
, 
E. Vendelova
E. Vendelova
 und 
S. Velebný
S. Velebný
   | 10. März 2016
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Article Category: Research Article
Online veröffentlicht: 10. März 2016
Seitenbereich: 3 - 13
Eingereicht: 30. Juli 2015
Akzeptiert: 16. Okt. 2015
DOI: https://doi.org/10.1515/helmin-2015-0062
Schlüsselwörter
© 2016 Institute of Parasitology, SAS, Košice
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1.

Microscopic immages of myelo-monocytic cells from the peritoneal cavities of mice with M. vogae infection. Peritoneal cells were isolated on selected days and cell smears were stained with May-Grünwald/Giemsa stains. (A) resident macrophage from non-infected mouse, (B) monocyte, day 7 p.i., (C) infl ammatory macrophages, day 35 p.i., (D) multinucleated “giant” macrophage, (E) macrophage attached to polymorphonuclear cell initiating its phagocytosis (F). Scale bar = 20 μm
Microscopic immages of myelo-monocytic cells from the peritoneal cavities of mice with M. vogae infection. Peritoneal cells were isolated on selected days and cell smears were stained with May-Grünwald/Giemsa stains. (A) resident macrophage from non-infected mouse, (B) monocyte, day 7 p.i., (C) infl ammatory macrophages, day 35 p.i., (D) multinucleated “giant” macrophage, (E) macrophage attached to polymorphonuclear cell initiating its phagocytosis (F). Scale bar = 20 μm

Fig. 2.

Proportion (%) of monocytes/macrophages/giant cells in the peritoneal cavities of non-infected and M. vogae infected ICR strain of mice. (A) proportions of macrophage–like cells from total isolated peritoneal exudate cells (PEC) within 5 weeks p.i., (B) proportion of morphologically distinct phenotypes of macrophages/monocytes/giant cells from total macrophage-like cells. Data shown for day 0 represent peritoneal cells from non-infected mice
Proportion (%) of monocytes/macrophages/giant cells in the peritoneal cavities of non-infected and M. vogae infected ICR strain of mice. (A) proportions of macrophage–like cells from total isolated peritoneal exudate cells (PEC) within 5 weeks p.i., (B) proportion of morphologically distinct phenotypes of macrophages/monocytes/giant cells from total macrophage-like cells. Data shown for day 0 represent peritoneal cells from non-infected mice

Fig. 3.

Phagocytosis of latex beads by peritoneal mononuclear phagocytes. (A) M. vogae-induced peritoneal cells were incubated with latex beads and mononuclear phagocyte populations were then assayed for phagocytosis by microscopy and expressed as percentage of phagocytic cells. (B) Peritoneal cells were pre-treated with ES (20 μg/ml), MvH (20 μg/ml) or medium and LPS as a control before addition of latex beads. Alternatively, latex beads were added to peritoneal cells together with LPS. Data represent mean ± SD of 4-5 independent in vitro experiments. *Significant difference (P <0.05) between data for naïve and post-infectious cells
Phagocytosis of latex beads by peritoneal mononuclear phagocytes. (A) M. vogae-induced peritoneal cells were incubated with latex beads and mononuclear phagocyte populations were then assayed for phagocytosis by microscopy and expressed as percentage of phagocytic cells. (B) Peritoneal cells were pre-treated with ES (20 μg/ml), MvH (20 μg/ml) or medium and LPS as a control before addition of latex beads. Alternatively, latex beads were added to peritoneal cells together with LPS. Data represent mean ± SD of 4-5 independent in vitro experiments. *Significant difference (P <0.05) between data for naïve and post-infectious cells

Fig. 4.

Microphotographs of monocytes/macrophages after incubation with latex beads (lxb) in vitro. Representative cells stained with May Grünwald/Giemsa stains that ingested lxb. (a) 0 lxb, (b) 2 lxb, (c) 5 lxb, (d) 8 lxb. Arrowhead indicates the ingested lxb (dull appearance), arrows indicate beads outside the cell (brighter, unstained). (x1000)
Microphotographs of monocytes/macrophages after incubation with latex beads (lxb) in vitro. Representative cells stained with May Grünwald/Giemsa stains that ingested lxb. (a) 0 lxb, (b) 2 lxb, (c) 5 lxb, (d) 8 lxb. Arrowhead indicates the ingested lxb (dull appearance), arrows indicate beads outside the cell (brighter, unstained). (x1000)

Fig. 5.

Phagocytosis/ endocytosis of [3H] cholesterol-labelled liposomes either non-opsonized or opsonised with serum components by peritoneal mononuclear phagocytes. Adherent monocytes/macrophages from non-infected (day 0 p.i.) and M. vogae-infected mice were incubated with untreated or serum pre-treated liposome suspensions (60 μg of liposomal lipids/well) and proportion of internalized particles was expressed as ng liposomal lipids/mg of cell proteins. (A) uptake of untreated liposomes by adherent monocytes/macrophages. **Significant difference (P <0.01) between uptake of naive and post-infectious phagocytes. (B) uptake of liposomes opsonised with normal mice serum (line A), with HI-normal mice serum (line B) and serum from mice with chronic M. vogae infection and without heat-inactivation (line C). Data are means ± SD of four in vitro tests on cells from 4 mice/day. Significant difference between treatment A and B, **(P <0.01), ***(P <0.001)
Phagocytosis/ endocytosis of [3H] cholesterol-labelled liposomes either non-opsonized or opsonised with serum components by peritoneal mononuclear phagocytes. Adherent monocytes/macrophages from non-infected (day 0 p.i.) and M. vogae-infected mice were incubated with untreated or serum pre-treated liposome suspensions (60 μg of liposomal lipids/well) and proportion of internalized particles was expressed as ng liposomal lipids/mg of cell proteins. (A) uptake of untreated liposomes by adherent monocytes/macrophages. **Significant difference (P <0.01) between uptake of naive and post-infectious phagocytes. (B) uptake of liposomes opsonised with normal mice serum (line A), with HI-normal mice serum (line B) and serum from mice with chronic M. vogae infection and without heat-inactivation (line C). Data are means ± SD of four in vitro tests on cells from 4 mice/day. Significant difference between treatment A and B, **(P <0.01), ***(P <0.001)

Phagocytosis of HEMA particles by peritoneal mononuclear phagocytes. Peritoneal cells isolated from non-infected mice and mice inoculated orally with M. vogae larvae were incubated with HEMA particles and mononuclear phagocyte populations were assayed ex vivo for phagocytic activity (FA) by microscopy, which was expressed as percentage of phagocytic cells from total counted cells. Index of phagocytic activity (IPA) was calculated as the average number of phagocytosed particles per phagocytosing cells from total particles counted. Data are mean ±SD after counting 300 cells per sample and assayed in duplicates for each mouse (n=8). Significantly different values from naïve cells isolated from non-infected mice (N): *** P<0.001; ** P<0.01; * P<0.05

Days p.i.N371014212835
FA53.3 ± 3.282.1 ± 4.8

P<0.001;

88.4 ± 5.9

P<0.001;

69.3 ± 3.5

P<0.01;

76.6 ± 2.9

P<0.001;

65.0 ± 1.8

P<0.05

61.2 ± 3.1

P<0.05

62.7 ± 2.8

P<0.05

IFA6.4 ± 0.910.6 ± 0.6

P<0.01;

12.1 ± 1.1

P<0.001;

6.8 ± 0.36.9 ± 0.47.1 ± 1.57.4 ± 0.9

P<0.05

6.2 ± 2.5
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