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Human parainfluenza virus type 2 (HPIV2) induced host ADAM8 expression in human salivary adenocarcinoma cell line (HSY) during cell fusion.

Ma GF, Miettinen S, Porola P, Hedman K, Salo J, Konttinen YT - BMC Microbiol. (2009)

Bottom Line: Without HPIV2 the corresponding percentages were only 7.7% and 8.8%.Moreover, ADAM8 positive cells formed bi- (16.2%) and multinuclear cells (3.5%) on day one and the corresponding percentages on day three were 15.6% for binuclear and 57.2% for multinuclear cells.The virus-HSY cell system provides a novel experimental model for study of the molecular mechanism of cell fusion events.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Medicine/invärtes medicin, Helsinki University Central Hospital, PO Box 700, Helsinki, Finland. guofeng.ma@helsinki.fi

ABSTRACT

Background: The aim of the study was to investigate expression of ADAMs (A Disintegrin and A Metalloproteinase) of host cell origin during cell-cell fusion induced by human parainfluenza virus type 2 (HPIV2).

Results: Induction of host cell ADAM9 was observed in GMK cells, but the applicability of this model was restricted by lack of cross-reactivity of the anti-human ADAM8 antibodies with the corresponding green monkey antigens. HSG cells were not susceptible to HPIV2 virus infection. In contrast, in human parotid gland HSY cells, a natural host cell for paramyxoviruses, HPIV2 induced ADAM8 expression. ADAM8 staining increased dramatically over time from 7.9 +/- 3% at zero hours to 99.2 +/- 0.8% at 72 hours (p = 0.0001). Without HPIV2 the corresponding percentages were only 7.7% and 8.8%. Moreover, ADAM8 positive cells formed bi- (16.2%) and multinuclear cells (3.5%) on day one and the corresponding percentages on day three were 15.6% for binuclear and 57.2% for multinuclear cells.

Conclusion: ADAM8, well recognized for participation in cell-to-cell fusion especially in osteoclast formation, is up-regulated upon formation of multinuclear giant cells after HPIV2 induction in HSY cells. The virus-HSY cell system provides a novel experimental model for study of the molecular mechanism of cell fusion events.

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Immunofluorescence double staining of ADAM8 and HPIV2 marker of HPIV2 stimulated HSY cell cultures on culture day 0 (panel B), 1 (panel C), 3 (panel D). ADAM8 staining is shown in red and HPIV2 shown in green (arrowheads), together with the blue nuclear counterstain of the same field. Panel A shows the staining of HSY cells that HPIV2 did not infect as negative control, therefore (A) only ADAM8 weak staining with nuclear counterstain, (B, C, D) overlay of double staining of ADAM8 and HPIV2 marker with blue nuclear counterstain of the same field on culture days 0, 1 and 3, respectively. Bar = 10 μm.
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Figure 2: Immunofluorescence double staining of ADAM8 and HPIV2 marker of HPIV2 stimulated HSY cell cultures on culture day 0 (panel B), 1 (panel C), 3 (panel D). ADAM8 staining is shown in red and HPIV2 shown in green (arrowheads), together with the blue nuclear counterstain of the same field. Panel A shows the staining of HSY cells that HPIV2 did not infect as negative control, therefore (A) only ADAM8 weak staining with nuclear counterstain, (B, C, D) overlay of double staining of ADAM8 and HPIV2 marker with blue nuclear counterstain of the same field on culture days 0, 1 and 3, respectively. Bar = 10 μm.

Mentions: In the uninfected HSY cells a very weak ADAM8 signal was seen (Figure 2A). At 2 hours HPIV2 was not yet found in HPIV2 infected HSY cell cultures and ADAM8 showed weak staining (Figure 2B). On culture day one, HPIV2 was seen inside HSY cells, which usually also showed cytoplasmic patches of immunoreactive ADAM8 (Figure 2C). On culture day three HPIV2 was found in some HSY cells. In addition, many large multinucleated cells were seen, which also were HPIV2 positive. In double label studies they stained for ADAM8, with a relatively strong signal, and a non-homogenous, granular and patchy cytoplasmic distribution (Figure 2D). In morphometric analysis, without HPIV2 stimulation the percentage of ADAM8 positive cells at 2 hours was 7.7 ± 0.9%, at 24 hours 7.5 ± 0.9% and at 72 hours 8.8 ± 1.0%. In HPIV2 infected cultures of human HSY cells the percentage of ADAM8 positive cells at 0 hour was 7.9 ± 3%, at 2 hours 15.0 ± 6.7% (p = 0.25), at 24 hours 57.0 ± 11% (p = 0.0719) and at 72 hours 99.2 ± 0.8% (p = 0.0001). All HPIV2 infected cells were also ADAM8 positive. We then calculated the percentages of ADAM8 and HPIV2 double positive cells and obtained that way also the number of ADAM8 positive but HPIV2 negative cells (Table 1). Moreover, ADAM8 positive cells formed also bi- and multinuclear cells. Fusion was seen already on day one at which time 16.2 ± 1.0% of the cells were binuclear and 3.5 ± 0.8% were multinuclear (all of them being ADAM8 positive). On day 3 15.6 ± 2.5% of the cells were binuclear (and all of them also ADAM8 positive) and altogether 57.2 ± 3.8% of all cells were multinuclear (and all of the also ADAM8 positive) (Figure 3).


Human parainfluenza virus type 2 (HPIV2) induced host ADAM8 expression in human salivary adenocarcinoma cell line (HSY) during cell fusion.

Ma GF, Miettinen S, Porola P, Hedman K, Salo J, Konttinen YT - BMC Microbiol. (2009)

Immunofluorescence double staining of ADAM8 and HPIV2 marker of HPIV2 stimulated HSY cell cultures on culture day 0 (panel B), 1 (panel C), 3 (panel D). ADAM8 staining is shown in red and HPIV2 shown in green (arrowheads), together with the blue nuclear counterstain of the same field. Panel A shows the staining of HSY cells that HPIV2 did not infect as negative control, therefore (A) only ADAM8 weak staining with nuclear counterstain, (B, C, D) overlay of double staining of ADAM8 and HPIV2 marker with blue nuclear counterstain of the same field on culture days 0, 1 and 3, respectively. Bar = 10 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2662866&req=5

Figure 2: Immunofluorescence double staining of ADAM8 and HPIV2 marker of HPIV2 stimulated HSY cell cultures on culture day 0 (panel B), 1 (panel C), 3 (panel D). ADAM8 staining is shown in red and HPIV2 shown in green (arrowheads), together with the blue nuclear counterstain of the same field. Panel A shows the staining of HSY cells that HPIV2 did not infect as negative control, therefore (A) only ADAM8 weak staining with nuclear counterstain, (B, C, D) overlay of double staining of ADAM8 and HPIV2 marker with blue nuclear counterstain of the same field on culture days 0, 1 and 3, respectively. Bar = 10 μm.
Mentions: In the uninfected HSY cells a very weak ADAM8 signal was seen (Figure 2A). At 2 hours HPIV2 was not yet found in HPIV2 infected HSY cell cultures and ADAM8 showed weak staining (Figure 2B). On culture day one, HPIV2 was seen inside HSY cells, which usually also showed cytoplasmic patches of immunoreactive ADAM8 (Figure 2C). On culture day three HPIV2 was found in some HSY cells. In addition, many large multinucleated cells were seen, which also were HPIV2 positive. In double label studies they stained for ADAM8, with a relatively strong signal, and a non-homogenous, granular and patchy cytoplasmic distribution (Figure 2D). In morphometric analysis, without HPIV2 stimulation the percentage of ADAM8 positive cells at 2 hours was 7.7 ± 0.9%, at 24 hours 7.5 ± 0.9% and at 72 hours 8.8 ± 1.0%. In HPIV2 infected cultures of human HSY cells the percentage of ADAM8 positive cells at 0 hour was 7.9 ± 3%, at 2 hours 15.0 ± 6.7% (p = 0.25), at 24 hours 57.0 ± 11% (p = 0.0719) and at 72 hours 99.2 ± 0.8% (p = 0.0001). All HPIV2 infected cells were also ADAM8 positive. We then calculated the percentages of ADAM8 and HPIV2 double positive cells and obtained that way also the number of ADAM8 positive but HPIV2 negative cells (Table 1). Moreover, ADAM8 positive cells formed also bi- and multinuclear cells. Fusion was seen already on day one at which time 16.2 ± 1.0% of the cells were binuclear and 3.5 ± 0.8% were multinuclear (all of them being ADAM8 positive). On day 3 15.6 ± 2.5% of the cells were binuclear (and all of them also ADAM8 positive) and altogether 57.2 ± 3.8% of all cells were multinuclear (and all of the also ADAM8 positive) (Figure 3).

Bottom Line: Without HPIV2 the corresponding percentages were only 7.7% and 8.8%.Moreover, ADAM8 positive cells formed bi- (16.2%) and multinuclear cells (3.5%) on day one and the corresponding percentages on day three were 15.6% for binuclear and 57.2% for multinuclear cells.The virus-HSY cell system provides a novel experimental model for study of the molecular mechanism of cell fusion events.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Medicine/invärtes medicin, Helsinki University Central Hospital, PO Box 700, Helsinki, Finland. guofeng.ma@helsinki.fi

ABSTRACT

Background: The aim of the study was to investigate expression of ADAMs (A Disintegrin and A Metalloproteinase) of host cell origin during cell-cell fusion induced by human parainfluenza virus type 2 (HPIV2).

Results: Induction of host cell ADAM9 was observed in GMK cells, but the applicability of this model was restricted by lack of cross-reactivity of the anti-human ADAM8 antibodies with the corresponding green monkey antigens. HSG cells were not susceptible to HPIV2 virus infection. In contrast, in human parotid gland HSY cells, a natural host cell for paramyxoviruses, HPIV2 induced ADAM8 expression. ADAM8 staining increased dramatically over time from 7.9 +/- 3% at zero hours to 99.2 +/- 0.8% at 72 hours (p = 0.0001). Without HPIV2 the corresponding percentages were only 7.7% and 8.8%. Moreover, ADAM8 positive cells formed bi- (16.2%) and multinuclear cells (3.5%) on day one and the corresponding percentages on day three were 15.6% for binuclear and 57.2% for multinuclear cells.

Conclusion: ADAM8, well recognized for participation in cell-to-cell fusion especially in osteoclast formation, is up-regulated upon formation of multinuclear giant cells after HPIV2 induction in HSY cells. The virus-HSY cell system provides a novel experimental model for study of the molecular mechanism of cell fusion events.

Show MeSH
Related in: MedlinePlus