Cell-surface heparan sulfate and HIV-1 infection of differentiated THP-1 cells

ORCiD

Nejat Düzgüneş: 0000-0001-6159-1391

Department

Biomedical Sciences

Document Type

Conference Presentation

Conference Title

82nd General Session of the International Association for Dental Research and 33rd Annual Meeting of the American Association for Dental Research

Location

Honolulu, HI

Conference Dates

March 10-13, 2004

Date of Presentation

3-12-2004

Journal Title

Journal of Dental Research

Journal ISSN

0022-0345

Journal Volume Number

83

First Page

2766

Abstract

Objectives: Recently, it has been reported that the cell surface glycosaminoglycan, heparan sulfate (HS), mediates the attachment of HIV to adherent cells expressing low CD4, such as HeLa-CD4 cells or macrophages, prior to virus entry. The established monocytic THP-1 cell line has been used to study HIV-monocyte/macrophage interactions and the relationship between differentiation, virus production and virus latency. Treatment with phorbol myristate acetate (PMA) induces differentiation of THP-1 cells into adherent macrophage-like cells, which are susceptible to M-tropic, CCR5-dependent HIV isolates. Differentiation of THP-1 cells markedly reduces CD4 surface expression (Konopka & Düzgünes, AIDS Res. Hum. Retroviruses 2002;18:123-131). Here we examined if HS is involved in HIV binding and infectivity in differentiated THP-1 cells. Methods: PMA-treated THP-1 (THP-1/PMA) cells were incubated with either heparinase (heparin-specific) or heparitinase I (HS-specific) for 2 h at 37°C. Expression of HS on the surface of THP-1 cells was examined using a fluorescent Monoclonal Anti-HS (10E4 epitope) antibody. Infection of THP-1 cells with the M-tropic HIV-1BaL isolate and HIV binding were monitored by ELISA determination of p24 antigen in harvested culture supernatants and cell lysates, respectively. Results: PMA-treatment resulted in an upregulation of HS expression. Over 40% of THP-1/PMA cells tested highly positive for HS expression (Mean Fluorescence Intensity, MFI = 22.5) when compared to 4.0% (MFI=5.4) for THP-1 cells growing in suspension. Treatment with the HS-specific heparitinase I reduced expression of HS by over 60%. Treatment with heparinases, however, did not reduce HIV binding and did not affect HIV infection in THP-1/PMA cells. Conclusions: Our results suggest that HIV infection in THP-1/PMA cells may be independent of cell-surface HS. (This work was supported partially by the Pacific Dental Research Foundation, Grant 521.)

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