Vaccines and immunotherapeutics derived from the human immunodeficiency virus (HIV) transactivator of transcription protein for the treatment and prevention of HIV disease
Anti-lentivirus vaccines and immunotherapeutics and methods for preparing and using same are disclosed. The vaccines and immunotherapeutics are produced using non-immunosuppressive lentivirus trans-activator of transcription (Tat) proteins. An associated in vitro ultra-sensitive macrophage Tat bioassay is disclosed for assessing the immunosuppressive qualities of the lentivirus Tat preparations of the present invention. Additionally, a related long-term T4 cell propagation system for characterizing lentivirus Tat is also disclosed. The present invention has additional utility in the treatment and prevention of AIDS.
1. A method of providing a long-term T4 cell propagation system comprising co-culturing uninfected peripheral blood mononuclear cells (PBMC) and PBMCs isolated from HIV-infected long-term non-progressors (LTNP); and diluting the co-culture to a very low density of 100 cells in 0.1 ml to 0.2 ml to initiate over-expression of immunostimulatory lentivirus trans-activator (IS-Tat) protein, wherein said very low cell density normally kills primary T4 cells.
2. The method of claim 1 , wherein the diluted co-culture density is 100 cells in 0.1 ml.
3. The method of claim 1 , wherein the diluted co-culture density is 100 cells in 0.2 ml.
4. The method of claim 1 , wherein said co-culture is diluted in 96-well tissue culture plates.