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Disinfection of cell-associated and extracellular HIV-1 by PUVA treatment
M Deichmann1, G Sczakiel, R Haas
1Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Abstract:
To inactivate cell-associated and extracellular HIV-1 while preserving cellular surface antigens, a procedure was used based on PUVA treatment, i.e. addition of psoralen to cell suspensions followed by irradiation with UVA light. T-lymphoid MT-4 cells were infected with HIV-1 strain NL4-3, 4'-aminomethyl-4,5',8-trimethylpsoralen was added, and the cell suspension was irradiated with 20 mW/cm2 UVA light for 3, 4 and 5 min. To evaluate virus inactivation, cells and supernatants were diluted serially and cocultured with uninfected MT-4 cells. Infectious HIV-1 was detected by cytopathic effects, immunofluorescence and p24 antigen ELISA. UVA irradiation at 3.6 J/cm2 (3 min 20 mW/cm2) reduced the amounts of both cell-associated and extracellular infectious HIV-1 by more than five orders of magnitude. Even at more stringent conditions of PUVA treatment (10 min 20 mW/cm2 UVA irradiation), conformational cellular surface epitopes remained detectable by flow cytometry.
Insights
PUVA treatment effectively inactivates cell-associated and extracellular HIV-1. This method preserves crucial cellular surface antigens, offering a promising approach for virus inactivation while maintaining cell integrity.
Area of Science:
- Virology
- Photobiology
- Cell Biology
Background:
- Human Immunodeficiency Virus type 1 (HIV-1) remains a significant global health challenge.
- Effective inactivation of both cell-associated and extracellular HIV-1 is crucial for therapeutic strategies and research.
- Preservation of cellular surface antigens during inactivation is vital for downstream applications.
Purpose of the Study:
- To evaluate the efficacy of PUVA treatment for inactivating HIV-1.
- To determine if PUVA treatment can preserve cellular surface antigens during virus inactivation.
- To establish optimal conditions for PUVA treatment of HIV-1.
Main Methods:
- Infection of T-lymphoid MT-4 cells with HIV-1 strain NL4-3.
- Application of 4'-aminomethyl-4,5',8-trimethylpsoralen followed by UVA irradiation.
- Assessment of viral inactivation through serial dilution, co-culture, cytopathic effect, immunofluorescence, and p24 antigen ELISA.
- Evaluation of cellular surface antigen preservation using flow cytometry.
Main Results:
- UVA irradiation at 3.6 J/cm2 (3 min) reduced infectious HIV-1 by over five orders of magnitude.
- Both cell-associated and extracellular infectious HIV-1 were significantly inactivated.
- Conformational cellular surface epitopes remained detectable by flow cytometry even under stringent PUVA conditions (10 min).
Conclusions:
- PUVA treatment is a highly effective method for inactivating cell-associated and extracellular HIV-1.
- The procedure successfully preserves cellular surface antigens, indicating its potential for safe handling and research.
- This photobiological inactivation method offers a promising strategy for HIV-1 management.