AEGiS-15IAC: Novel method of Inactivation of HIV-1 by the freeze pressure generation method (FPGM).

15th International AIDS Conference

Bangkok, Thailand - July 11-16, 2004

Novel method of Inactivation of HIV-1 by the freeze pressure generation method (FPGM).

Int Conf AIDS 2004 Jul 11-16; 15:(abstract no. C10237)

Otake T, Kawahata T, Mori H, Kojima Y, Hayakawa K
Osaka Prefectural Institute of Public Health, Osaka, Japan

BACKGROUND: HIV infection through blood transfusion is still possible if donated blood collected during the so-called window period is used for the transfusion. We already reported that high-pressure (over 600 MPa) treatment at room temperature inactivates HIV-1, and have recently shown that high pressure generated by the expansion of water due to freezing (freeze pressure generation method, FPGM) has an inactivating effect on bacteria.

METHODS: Seven HIV-1 strains were used: 4 T cell-tropic strains and 3 macrophage-tropic strains including clinical isolates. A plastic tube containing a virus suspension was placed in a pure-water-filled stainless pressure-tight container, which was left in a freezer for 24 hours. The infective titer of the virus was determined using MT-4 and MAGIC5A cells. Reverse transcriptase (RT) activity was measured using a nonradioisotope RT activity assay kit. In addition, the viral envelope function was tested. A concentrated virus sample was reacted with MT-4 cells, which were reacted with anti-gp120 antibody to measure the amount of virus bound to the cells.

RESULTS: The virus infective titer of all HIV-1 strains decreased to approximately 1/100 after treatment at -10degreesC (100MPa), and below the detection limit after treatment at -20degreesC (200 MPa) or -30degreesC (250 MPa). The RT activity of HIV-1 remained unchanged at -10degreesC, but decreased to approximately 1/10 at -20degreesC or -30degreesC. In addition, we confirmed that the ability of the virus to bind to the surface of cells was lost by treatment at -20degreesC.

CONCLUSIONS: Several HIV-1 strains including clinical isolates became completely inactivated by FPGM at -20degreesC (200 MPa) to -30degreesC (250 MPa). The mechanism of the virus inactivation may involve virus enzyme damage and changes in virus envelope proteins. FPGM seemed to be simple and convenient method for the detoxication of blood preparation.

Keywords: AEGIS, HIV-1, HIV Envelope Protein gp120, RNA-Directed DNA Polymerase, HIV Infections, Pressure, Virion, Macrophages, T-Lymphocytes, immunology, methods

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C10237