Inhibition of HIV-1 Nef-induced apoptosis of uninfected human blood cells by serine/threonine protein kinase inhibitors, fasudil hydrochloride and M3.
The Nef protein of HIV-1 binds to and induces apoptotic cytolysis of uninfected but activated human peripheral blood mononuclear cells (PBMC) and various cell line cells derived from CD4+ T, CD8+ T and B lymphocytes, macrophages, and neutrophils. The Nef-induced apoptosis also occurs with blood cells not expressing CD95 (Fas). The Nef-induced apoptosis as well as Fas-mediated apoptosis was inhibited by acetyl-Try-Val-Ala-Asp-CHO, an IL-1beta converting enzyme (ICE) inhibitor. On the other hand, serine/threonine protein kinase (PK) inhibitors, H-7, fasudil hydrochloride and M3, inhibited the Nef-induced apoptosis, and not the Fas-mediated one, without affecting the cell-binding activity of Nef and Nef-binding capacity of the activated cells. Preincubation of the cells with the drugs before being bound by Nef was required for the inhibition of apoptosis. These results suggest that the PK inhibitors specifically act on a cellular protein involved in the upper stream of signal transduction pathway of the Nef-induced apoptosis, which is different from the Fas-mediated pathway but meets it upstream of ICE. In addition, the drugs suppressed the cellular activation-associated cell surface expression of a putative Nef-binding protein in PBMC, although they had no influence on its expression in cell line cells. These findings suggest the feasibility of clinical use of the PK inhibitors to prevent the development of AIDS by inhibiting the Nef-induced apoptosis of uninfected blood cells.
Okada
H
H
Department of Virology 1, National Institute of Infectious Diseases, Tokyo, Japan.
Takei
R
R
Tashiro
M
M
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000163:Acquired Immunodeficiency Syndrome / Q000517:prevention & control*; D000328:Adult; D019380:Anti-HIV Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005240:Feasibility Studies; D015693:Gene Products, nef / Q000037:antagonists & inhibitors*; D015497:HIV-1 / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors*; D015728:Receptors, HIV / Q000378:metabolism; D013601:T-Lymphocytes; D014407:Tumor Cells, Cultured; D019014:fas Receptor / Q000378:metabolism; D054311:nef Gene Products, Human Immunodeficiency Virus
C110851:1-(1-hydroxy-5-isoquinolinesulfonyl)homopiperazine; D019380:Anti-HIV Agents; D004791:Enzyme Inhibitors; D015693:Gene Products, nef; C110852:Nef receptor; D015728:Receptors, HIV; D019014:fas Receptor; D054311:nef Gene Products, Human Immunodeficiency Virus; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D017346:Protein Serine-Threonine Kinases; C049347:fasudil