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V beta activation by HIV Nef protein: detection by a simple amplification procedure
T Tanabe1, B A Torres, P S Subramaniam
1Department of Microbiology and Cell Science, University of Florida, Gainesville 32611, USA.
Biochemical and Biophysical Research Communications
|January 23, 1997
Summary
Human immunodeficiency virus (HIV) Nef protein weakly stimulates T-cells. A new method detects this specific T-cell expansion, identifying V beta 5.3 and V beta 18 T-cells in treated cultures.
Area of Science:
- Immunology
- Virology
Background:
- Human immunodeficiency virus (HIV) Nef protein exhibits superantigen properties.
- Previous attempts to demonstrate Nef-induced V beta-specific T-cell expansion via flow cytometry were inconsistent due to Nef's lower mitogenic activity.
Purpose of the Study:
- To develop a simple amplification detection method for identifying V beta-specific T-cell expansion induced by weak superantigens like HIV Nef.
- To confirm the superantigen activity of HIV Nef on specific T-cell subsets.
Main Methods:
- Human peripheral blood mononuclear cells (PBMCs) were treated with HIV Nef protein.
- Cells were restimulated using immobilized anti-V beta antibodies.
- T-cell expansion was detected using the novel amplification method and confirmed with reverse transcriptase-polymerase chain reaction (RT-PCR).
Main Results:
- Significant expansion of V beta 5.3 and V beta 18 T-cells was detected in Nef-treated cultures.
- V beta 18 expansion was observed in all donors, while V beta 5.3 expansion occurred in 50% of donors.
- Variable responses were noted for V beta 2, V beta 3, and V beta 9 T-cells, consistent with RT-PCR results.
Conclusions:
- A novel, reproducible, and simple amplification detection method effectively identifies V beta-specific T-cell expansion induced by weak superantigens.
- HIV Nef protein demonstrates superantigen activity, inducing expansion of specific T-cell subsets, particularly V beta 18 and V beta 5.3.