Abstract
We previously reported that Nef protein from human immunodeficiency virus (HIV) has superantigen properties. However,we were unable to consistently demonstrate Vβ-specific expansion by Nef using flow cytometry, possibly due to its lower mitogenic activity compared to prototypic superantigens. Therefore, we developed a simple amplification detection method using immobilized anti-Vβ antibodies. Human peripheral blood mononuclear cells from adult donors were treated for 24 h with Nef and restimulated with immobilized anti-Vβ antibodies for an additional 72 h. Significant expansion of Vβ5.3 and Vβ18 T-cells were detected in Nef-treated cultures, with expansion of Vβ18 occurring with all donors tested, and Vβ5.3 expansion occurring in 50% of the donors. Variable responses were obtained with Vβ2, Vβ3, and Vβ9. These results were confirmed using the more time-consuming method of reverse transcriptase-polymerase chain reaction (RT-PCR). Thus, this novel, reproducible, and relatively simple method can detect Vβ-specific expansion by weak superantigens.
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