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The novel surfactant protein SP-H enhances the phagocytosis efficiency of macrophage-like cell lines U937 and MH-S.

Diler E, Schicht M, Rabung A, Tschernig T, Meier C, Rausch F, Garreis F, Bräuer L, Paulsen F - BMC Res Notes (2014)

Bottom Line: SP-H markedly increases phagocytosis in vitro in the murine-derived alveolar macrophage cell lines MH-S and in human-derived differentiated U937 cells.It can be assumed that SP-H is involved in regulating phagocytic activity of macrophages.SP-H is a new player in pulmonary host defence.

View Article: PubMed Central - PubMed

Affiliation: Institute of Anatomy and Cell Biology, Saarland University, 66421 Homburg/Saar, Germany. thomas.tschernig@uks.eu.

ABSTRACT

Background: Surfactant proteins (SP) secreted by alveolar type 2 cells, play an essential role in maintaining the air-liquid barrier of the lung and are also involved in the opsonisation and clearance of bacteria by phagocytes. We have recently described a novel surfactant protein, SP-H (SFTA3). Expression of SP-H was earlier demonstrated to be upregulated by LPS and negatively regulated by IL-1β and IL-23 in vitro. The influence of SP-H on phagocytosis was measured using a murine and a human phagocytic cell line and fluorescent latex beads.

Findings: SP-H markedly increases phagocytosis in vitro in the murine-derived alveolar macrophage cell lines MH-S and in human-derived differentiated U937 cells.

Conclusion: It can be assumed that SP-H is involved in regulating phagocytic activity of macrophages. SP-H is a new player in pulmonary host defence.

No MeSH data available.


Related in: MedlinePlus

The phagocytosis efficiency of the alveolar macrophage cell line MH-S and the differentiated lymphoma cell line U937 as a function of SP-H. The phagocytosis efficiency was determined by flow cytometry.
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Fig1: The phagocytosis efficiency of the alveolar macrophage cell line MH-S and the differentiated lymphoma cell line U937 as a function of SP-H. The phagocytosis efficiency was determined by flow cytometry.

Mentions: The results revealed that the efficiency of bead uptake by both cell lines was significantly enhanced by the presence of the SP-H protein. The gradually increasing concentration of the protein also caused a gradual increase in phagocytosis efficiency. The results show that, in the presence of 500 ng ml-1 and 1 μg ml-1 SP-H, both cell lines are significantly stimulated to take up particles (Figure 1). Obviously, the murine alveolar macrophage cell line MH-S has a higher susceptibility to SP-H than the human U937, which was initially isolated from histocytic lymphoma [10]. In summary, the findings of this study demonstrate an improving effect of SP-H on the phagocytosis of latex particles by macrophage like cell lines of human and of mouse origin. Especially the alveolar macrophage cell line MH-S is highly stimulated by SP-H, even at a concentration of 100 ng ml-1, whereas the differentiated U937 cells showed only slightly increased phagocytosis efficiency at a five-fold higher concentration (500 ng ml-1). To exclude that an increase in the relative fluorescence intensity (RFI) measured by flow cytometer is caused by an enhanced adherence of the polystyrene microspheres to the cells, the assays were performed after incubation of the cells with phagocytosis inhibitors. Figure 2 shows that in the presence of the inhibitors, SP-H did not have a phagocytosis efficiency increasing effect on MH-S cells. In contrast, U937 cells treated with phagocytosis inhibitors, showed a significant increase in their phagocytosis efficiency caused by 1 μg ml-1 SP-H. However, the phagocytosis of the U937 cells with 1 μg ml-1 SP-H treated with inhibitors is significantly lower than the U937 cells that were not treated with inhibitors (Figure 3). Therefore, the increase in RFI values are attributable to an enhanced phagocytosis efficiency, rather than to improved adhesion of the beads to the cells.Figure 1


The novel surfactant protein SP-H enhances the phagocytosis efficiency of macrophage-like cell lines U937 and MH-S.

Diler E, Schicht M, Rabung A, Tschernig T, Meier C, Rausch F, Garreis F, Bräuer L, Paulsen F - BMC Res Notes (2014)

The phagocytosis efficiency of the alveolar macrophage cell line MH-S and the differentiated lymphoma cell line U937 as a function of SP-H. The phagocytosis efficiency was determined by flow cytometry.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4256058&req=5

Fig1: The phagocytosis efficiency of the alveolar macrophage cell line MH-S and the differentiated lymphoma cell line U937 as a function of SP-H. The phagocytosis efficiency was determined by flow cytometry.
Mentions: The results revealed that the efficiency of bead uptake by both cell lines was significantly enhanced by the presence of the SP-H protein. The gradually increasing concentration of the protein also caused a gradual increase in phagocytosis efficiency. The results show that, in the presence of 500 ng ml-1 and 1 μg ml-1 SP-H, both cell lines are significantly stimulated to take up particles (Figure 1). Obviously, the murine alveolar macrophage cell line MH-S has a higher susceptibility to SP-H than the human U937, which was initially isolated from histocytic lymphoma [10]. In summary, the findings of this study demonstrate an improving effect of SP-H on the phagocytosis of latex particles by macrophage like cell lines of human and of mouse origin. Especially the alveolar macrophage cell line MH-S is highly stimulated by SP-H, even at a concentration of 100 ng ml-1, whereas the differentiated U937 cells showed only slightly increased phagocytosis efficiency at a five-fold higher concentration (500 ng ml-1). To exclude that an increase in the relative fluorescence intensity (RFI) measured by flow cytometer is caused by an enhanced adherence of the polystyrene microspheres to the cells, the assays were performed after incubation of the cells with phagocytosis inhibitors. Figure 2 shows that in the presence of the inhibitors, SP-H did not have a phagocytosis efficiency increasing effect on MH-S cells. In contrast, U937 cells treated with phagocytosis inhibitors, showed a significant increase in their phagocytosis efficiency caused by 1 μg ml-1 SP-H. However, the phagocytosis of the U937 cells with 1 μg ml-1 SP-H treated with inhibitors is significantly lower than the U937 cells that were not treated with inhibitors (Figure 3). Therefore, the increase in RFI values are attributable to an enhanced phagocytosis efficiency, rather than to improved adhesion of the beads to the cells.Figure 1

Bottom Line: SP-H markedly increases phagocytosis in vitro in the murine-derived alveolar macrophage cell lines MH-S and in human-derived differentiated U937 cells.It can be assumed that SP-H is involved in regulating phagocytic activity of macrophages.SP-H is a new player in pulmonary host defence.

View Article: PubMed Central - PubMed

Affiliation: Institute of Anatomy and Cell Biology, Saarland University, 66421 Homburg/Saar, Germany. thomas.tschernig@uks.eu.

ABSTRACT

Background: Surfactant proteins (SP) secreted by alveolar type 2 cells, play an essential role in maintaining the air-liquid barrier of the lung and are also involved in the opsonisation and clearance of bacteria by phagocytes. We have recently described a novel surfactant protein, SP-H (SFTA3). Expression of SP-H was earlier demonstrated to be upregulated by LPS and negatively regulated by IL-1β and IL-23 in vitro. The influence of SP-H on phagocytosis was measured using a murine and a human phagocytic cell line and fluorescent latex beads.

Findings: SP-H markedly increases phagocytosis in vitro in the murine-derived alveolar macrophage cell lines MH-S and in human-derived differentiated U937 cells.

Conclusion: It can be assumed that SP-H is involved in regulating phagocytic activity of macrophages. SP-H is a new player in pulmonary host defence.

No MeSH data available.


Related in: MedlinePlus