Inhibition of the Protease of Human Immunodeficiency Virus Blocks Replication and Infectivity of the Virus in Chronically Infected Macrophages

Abstract

Because of the importance of monocytes/macrophages (M/M) as an in vivo reservoir of human immunodeficiency virus (HIV), a study was done to investigate whether viral replication in chronically infected macrophages (HIV M/M) could be inhibited by various drugs, including U-75875, an inhibitor of HIV protease. HIV replication in M/M and in chronically infected T cells was dramatically decreased by U-75875, while other drugs, including zidovudine, interferon-alpha, and an antisense oligodeoxynucleotide against the rev gene, were effective antiviral agents only in de novo-infected cells. Virus titer in HIV M/M was reduced approximately 10(5)-fold by nontoxic concentrations of U-75875, while no effect on HIV DNA or virus antigen expression on cell membrane was achieved in M/M infected either chronically or de novo. Thus, U-75875 essentially worked against late stages of viral replication. These data support the use of protease inhibitors, alone or in combination, in the therapy of HIV-infected patients.

Human Immunodeficiency Virus Type 1 Infection Modulates the Interleukin (IL)–1β and IL‐6 Responses of Human Macrophages to CD40 Ligand Stimulation

“…Brefeldin A, saponin, lipopolysaccharide (LPS) from Escherichia coli O111:B4, polymyxin B, and bovine serum albumin were from Sigma (St. Louis). U75875, a synthetic peptidomimetic inhibitor of the HIV-1 protease [21], was supplied by Upjohn Laboratories (Kalamazoo, MI); U75875 is for laboratory use only and is not to be administered to humans or food-producing animals or plants. Zidovudine was obtained from Sigma (Milan).…”

Comparative antiviral activity of integrase inhibitors in human monocyte-derived macrophages and lymphocytes

“…Human primary M/M were prepared and purified as described in published procedures (Bagnarelli et al, 1996;Cenci et al, 1997;Perno et al, 1998). Briefly, M/M were obtained from the blood of healthy HIV-seronegative donors.…”

“…These latently infected cells could expand the viral reservoir continuously and render the infection impossible to eradicate from the body in the absence of specific pharmacological interventions (Chomont et al, 2011;Dieffenbach and Fauci, 2011;Pomerantz, 2001;Pomerantz and Horn, 2003). Infected macrophages, which can survive and produce virus for long periods, play an important role in all phases of HIV-1 infection and in various aspects of the disease process, acting as a vehicle for virus dissemination, and damage to bystander cells and therefore representing another major and important obstacle to the complete eradication of virus even in the presence of antiretroviral therapy (Alexaki et al, 2008;Aquaro et al, 1997Aquaro et al, , 1998Aquaro et al, , 2002aAquaro and Perno, 2005;Coleman and Wu, 2009;Herbein et al, 2010;McGrath, 1996;Sharova et al, 2005).…”

Novel In Vivo Model for the Study of Human Immunodeficiency Virus Type 1 Transcription Inhibitors: Evaluation of New 6-Desfluoroquinolone Derivatives

“…A monocytotropic HIV-1 strain was used for studies involving primary M/M. The characteristics and genomic sequence of this strain have been described previously (12,18,36,37). The virus was incubated with M/M, and supernatants were collected, filtered, and stored at Ϫ80°C before use (36).…”

“…The characteristics and genomic sequence of this strain have been described previously (12,18,36,37). The virus was incubated with M/M, and supernatants were collected, filtered, and stored at Ϫ80°C before use (36). The characteristics of viral stocks used for this study were 2.1 ϫ 10 8 HIV RNA genomes/ml (corresponding to 35 ng/ml of p24 antigen) and 5 ϫ 10 3 50% tissue culture infective doses per ml, as assessed by virus titration in other primary M/M cultures.…”