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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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Related in: MedlinePlus

Immunofluorescence double staining of ADAM9 and HPIV2 marker of HPIV2 stimulated GMK cell cultures on culture day 0 (panel A, B), 1 (panel C), 3 (panel D). ADAM9 staining is shown in red and HPIV2 shown in green, together with the blue nuclear counterstain of the same field. On culture day 0 HPIV2 did not infect any cells, therefore (A) shows only nuclear counterstain, (B) shows only ADAM9 staining, (C, D) overlays show double staining of ADAM9 and HPIV2 marker on culture days 1 and 3, respectively. Bar = 10 μm.
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Figure 1: Immunofluorescence double staining of ADAM9 and HPIV2 marker of HPIV2 stimulated GMK cell cultures on culture day 0 (panel A, B), 1 (panel C), 3 (panel D). ADAM9 staining is shown in red and HPIV2 shown in green, together with the blue nuclear counterstain of the same field. On culture day 0 HPIV2 did not infect any cells, therefore (A) shows only nuclear counterstain, (B) shows only ADAM9 staining, (C, D) overlays show double staining of ADAM9 and HPIV2 marker on culture days 1 and 3, respectively. Bar = 10 μm.

Mentions: At 2 hours HPIV2 antigens were not yet found in infected GMK cells (data not shown) and ADAM9 was absent (Figure 1A, B). On culture day 1 HPIV2 was seen in infected GMK cells and all the infected and some of the uninfected GMK cells were ADAM9 positive (Figure 1C). On culture day 3 HPIV2 had infected most GMK cells and had caused cytopathic effects including formation of large multinucleated syncytia. The multinuclear giant cells were relatively strongly labeled for ADAM9 (Figure 1D). The positive controls of ADAMs were positive showing that the immunolabeling protocol used worked acceptably; also the negative staining controls were negative showing that the ADAM9 staining results were correctly positive (data not shown).


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 ADAM9 and HPIV2 marker of HPIV2 stimulated GMK cell cultures on culture day 0 (panel A, B), 1 (panel C), 3 (panel D). ADAM9 staining is shown in red and HPIV2 shown in green, together with the blue nuclear counterstain of the same field. On culture day 0 HPIV2 did not infect any cells, therefore (A) shows only nuclear counterstain, (B) shows only ADAM9 staining, (C, D) overlays show double staining of ADAM9 and HPIV2 marker on culture days 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 1: Immunofluorescence double staining of ADAM9 and HPIV2 marker of HPIV2 stimulated GMK cell cultures on culture day 0 (panel A, B), 1 (panel C), 3 (panel D). ADAM9 staining is shown in red and HPIV2 shown in green, together with the blue nuclear counterstain of the same field. On culture day 0 HPIV2 did not infect any cells, therefore (A) shows only nuclear counterstain, (B) shows only ADAM9 staining, (C, D) overlays show double staining of ADAM9 and HPIV2 marker on culture days 1 and 3, respectively. Bar = 10 μm.
Mentions: At 2 hours HPIV2 antigens were not yet found in infected GMK cells (data not shown) and ADAM9 was absent (Figure 1A, B). On culture day 1 HPIV2 was seen in infected GMK cells and all the infected and some of the uninfected GMK cells were ADAM9 positive (Figure 1C). On culture day 3 HPIV2 had infected most GMK cells and had caused cytopathic effects including formation of large multinucleated syncytia. The multinuclear giant cells were relatively strongly labeled for ADAM9 (Figure 1D). The positive controls of ADAMs were positive showing that the immunolabeling protocol used worked acceptably; also the negative staining controls were negative showing that the ADAM9 staining results were correctly positive (data not shown).

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