Glycobiology Advance Access originally published online on October 8, 2008
Glycobiology 2009 19(1):76-82; doi:10.1093/glycob/cwn106
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Induction of HIV-1 resistance: cell susceptibility to infection is an inverse function of globotriaosyl ceramide levels
2 Department of Laboratory Medicine and Pathobiology, University of Toronto
3 Research and Development of Canadian Blood Services, Toronto, Ontario M5G 2M1
4 Department of Medicine, University of Toronto
5 Division of Cell and Molecular Biology, Toronto General Research Institute of the University Health Network
6 Research Institute, Hospital for Sick Children, Toronto, Ontario M5G 1X8
7 Department of Biochemistry, University of Toronto, Canada
1 To whom correspondence should be addressed: Tel: +1-416-813-5998; Fax: +416-813-5993; e-mail: cling{at}sickkids.ca
Received on August 1, 2008; revised on August 29, 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. The 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 100 µM. In contrast, P4 (2 µM) 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 / gp120 / lipid rafts / pk blood group antigen
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  R. Mahfoud, A. Manis, and C. A. Lingwood Fatty acid-dependent globotriaosyl ceramide receptor function in detergent resistant model membranes J. Lipid Res., September 1, 2009; 50(9): 1744 - 1755. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Suzuki, T.-H. Su, S.-W. Wu, K. Yamamoto, K.-H. Khoo, and Y. C Lee Structural analysist of N-glycans from gull egg white glycoproteins and egg yolk IgG Glycobiology, July 1, 2009; 19(7): 693 - 706. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Lund, M. L. Olsson, S. Ramkumar, D. Sakac, V. Yahalom, C. Levene, A. Hellberg, X.-Z. Ma, B. Binnington, D. Jung, et al. The human Pk histo-blood group antigen provides protection against HIV-1 infection Blood, May 14, 2009; 113(20): 4980 - 4991. [Abstract] [Full Text] [PDF]
|
|