Anti-HIV activity of newly cloned secretory leukocyte protease inhibitor (SLPI) in human macrophages

ORCiD

Nejat Düzgüneş: 0000-0001-6159-1391

Department

Biomedical Sciences

Document Type

Conference Presentation

Conference Title

32nd Annual Meeting of the American Association for Dental Research

Location

San Antonio, TX

Conference Dates

March 12-15, 2003

Date of Presentation

3-13-2003

Journal Title

Journal of Dental Research

Journal ISSN

0022-0345

Journal Volume Number

82 (special issue A)

First Page

504

Abstract

Objectives: Secretory leukocyte protease inhibitor (SLPI), a serine protease inhibitor present in saliva, has been shown to inhibit HIV infection of macrophages and primary T-cells (McNeely et al., J. Clin. Invest. 1995; 96:456; Blood 1997; 90:1141; Shugars et al., Oral. Dis. 1997; 3 (Suppl.1):S70). However, contradictory results have also been published (Turpin et al., Antiviral Res. 1996; 29:269) and variable results were observed in our laboratory (Konopka et al., J. Dent. Res. 1999; 78: 1773). All these studies have been performed using the same, commercially available, preparation of recombinant (r) SLPI obtained from Synergen and/or R&D Systems. In this study we examined the anti-HIV activity of a newly cloned SLPI using monocyte-derived macrophages. Methods: The SLPI gene was synthesized by a PCR-based strategy using long oligonucleotides, and the protein was overexpressed in E. coli, purified, and successfully refolded. Macrophages were infected with HIV-1Ba-L on day 10 or 14 post-isolation, in the absence or presence of SLPI. Cells were incubated with SLPI for 30 min at 37°C prior to addition of the virus. SLPI was also present during the 2 h infection period. Virus production in culture supernatants was monitored by p24 ELISA. Results: Over 90% inhibition of p24 production was observed in macrophages infected in the presence of 5, 10 or 20 µg/ml of SLPI. At 0.5 or 1 µg/ml, SLPI reduced the p24 levels by ~70 to 80%. This potent inhibition of virus production was sustained for 2 weeks of culture. Conclusions: (i) the newly cloned SLPI effectively blocks HIV-1 infection in macrophages, and (ii) the availability of an active rSLPI will enable the investigation of the biochemical and structural basis for the anti-HIV activity of SLPI. Partially supported by Pacific Dental Research Foundation Grant 518, and funds from the University of the Pacific School of Dentistry.

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