Glycobiology Advance Access published online on October 8, 2008
Glycobiology, doi:10.1093/glycob/cwn106
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Induction of HIV-1 resistance: Cell susceptibility to infection is an inverse function of globotriaosyl ceramide levels
1 Department of Laboratory Medicine and Pathobiology, University of Toronto, Canada
2 Research and Development of Canadian Blood Services, Toronto, Ontario M5G 2M1 Canada
3 Department of Medicine, University of Toronto, Canada
4 Division of Cell and Molecular Biology, Toronto General Research Institute of the University Health Network, Canada
5 Research Institute, Hospital for Sick Children, Toronto, Ontario M5G 1x8 Canada
6 Department of Biochemistry, University of Toronto, Canada
Correspondence to C.A. Lingwood, Hospital for Sick Children, Toronto, Ontario M5G 1x8, CANADA, Tel: 416–813-5998, Email: cling{at}sickkids.ca
Received on August 1, 2008; accepted on October 1, 2008
To examine the role of the glycosphingolipid (GSL), globotriaosylceramide (Gb3, CD77, pk blood group antigen) in HIV-1 infection, we have pharmacologically modulated Gb3 metabolism in an X4 HIV-1 infectable monocytic cell line (THP-1) that naturally expresses Gb3 and in a Gb3 expressing glioblastoma cell line (U87) transfected to express both CD4 and CCR5 to permit R5 HIV-1 infection.
THP-1 and U87 cells were treated with either a competitive inhibitor of 
-galactosidase A, 1-deoxygalactonojirimycin (DGJ) to induce Gb3 accumulation, or a glucosylceramide synthase inhibitor, phenyl -2- palmitylamino-3- pyrrolidino -1-propanol (P4) to deplete cells of Gb3. HIV susceptibility was determined via measurement of p24gag antigen production by ELISA. In addition, total cellular Gb3 content was determined using thin layer chromatography followed by Verotoxin1 overlay binding. Cell surface expression of Gb3 was verified by FACS analysis.
We found that DGJ significantly decreased THP-1 and U87 cell susceptibility to HIV-1IIIB and HIV-1BaL infection respectively at a concentration of approximately 100µM. In contrast, P4 (2µM) substantially increased cellular susceptibility to HIV-1 infection. Total cellular GSL analysis verified increased Gb3 expression in cells treated with DGJ and considerable reduction of Gb3 in P4-treated cells as compared to controls. These results show a reciprocal relationship between Gb3 expression and infection with either X4 HIV-1IIIB or R5 HIV-1Ba-L.
These results support previous studies that Gb3 provides resistance to HIV infection. Variable Gb3 expression may provide a natural HIV resistance factor in the general population and pharmacological manipulation of Gb3 levels may provide an approach to induction of HIV resistance.
Key words: glycosphingolipids / lipid rafts / gp120 / pk blood group antigen
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