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J. Virol., Mar 1995, 1462-1472, Vol 69, No. 3
S Frey, M Marsh, S Gunther, A Pelchen-Matthews, P Stephens, S Ortlepp and T Stegmann
We investigated cell-cell fusion induced by the envelope glycoprotein of
human immunodeficiency virus type 1 strain IIIB expressed on the surface of
CHO cells. These cells formed syncytia when incubated together with
CD4-positive human lymphoblastoid SupT1 cells or HeLa-CD4 cells but not
when incubated with CD4-negative cell lines. A new assay for binding and
fusion was developed by using fluorescent phospholipid analogs that were
produced in SupT1 cells by metabolic incorporation of BODIPY-labeled fatty
acids. Fusion occurred as early as 10 min after mixing of labeled SupT1
cells with unlabeled CHO-gp160 cells at 37 degrees C. When both the
fluorescence assay and formation of syncytia were used, fusion of SupT1 and
HeLa-CD4 cells with CHO-gp160 cells was observed only at temperatures above
25 degrees C, confirming recent observations (Y.-K. Fu, T.K. Hart, Z.L.
Jonak, and P.J. Bugelski, J. Virol. 67:3818-3825, 1993). This temperature
dependence was not observed with influenza virus-induced cell-cell fusion,
which was quantitatively similar at both 20 and 37 degrees C, indicating
that cell-cell fusion in general is not temperature dependent in this
range. gp120-CD4-specific cell-cell binding was found over the entire 0 to
37 degrees C range but increased markedly above 25 degrees C. The enhanced
binding and fusion were reduced by cytochalasins B and D. Binding of
soluble gp120 to CD4-expressing cells was equivalent at 37 and 16 degrees
C. Together, these data indicate that during gp120-gp41-induced syncytium
formation, initial cell-cell binding is followed by a
cytoskeleton-dependent increase in the number of gp120-CD4 complexes,
leading to an increase in the avidity of cell-cell binding. The increased
number of gp120-CD4 complexes is required for fusion, which suggests that
the formation of a fusion complex consisting of multiple CD4 and gp120-gp41
molecules is a step in the fusion mechanism.
Copyright © 1995, American Society for Microbiology
Temperature dependence of cell-cell fusion induced by the envelope glycoprotein of human immunodeficiency virus type 1
Department of Biophysical Chemistry, Biozentrum of the University of Basel, Switzerland.
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