Effects of MS-8209, an Amphotericin B Derivative, on Tumor Necrosis Factor Alpha Synthesis and Human Immunodeficiency Virus Replication in Macrophages

Abstract

Amphotericin B derivatives, such as MS-8209, have been evaluated as a therapeutic approach to human immunodeficiency virus (HIV) infection. We show that MS-8209, like amphotericin B, increases tumor necrosis factor alpha (TNF-␣) mRNA expression and TNF-␣ production and consequently HIV replication in human macrophages. These effects confirm the pharmacological risk associated with the administration of amphotericin B or its derivatives to HIV-infected patients.Macrophages and related cells play a key role in pathological events, particularly in inflammatory processes associated with human immunodeficiency virus (HIV) disease. They are a major target cell for HIV, and proinflammatory monokines such as tumor necrosis factor alpha (TNF-␣) increase HIV type 1 (HIV-1) gene expression and replication, via intracellular activation and autocrine and paracrine pathways (5,14,19). MS-8209 is a water-soluble derivative of amphotericin B (AmB) of lower cellular and animal toxicity and greater solubility than the parent compound (13). MS-8209 exhibits antiretroviral activity in mitogen-activated CD4 ϩ T lymphocytes infected in vitro with HIV-1 (4, 11). MS-8209 exerts its antiviral action by inhibiting HIV entry into cells after CD4-gp120 interactions (13). AmB provokes marked overexpression of TNF-␣ in murine and human macrophages (10,15,17). In view of the deleterious effects of TNF-␣ on HIV replication, it was therefore of interest to investigate the possible effects of MS-8209 on TNF-␣ synthesis and HIV replication in macrophages.We obtained human macrophages by 7-day differentiation of freshly isolated monocytes. Monocytes were separated from peripheral blood mononuclear cells using countercurrent centrifugal elutriation. Cells were cultured in RPMI 1640 supplemented with 10% heat-inactivated fetal calf serum, 2 mM L-glutamine, and 1% triantibiotic mixture (penicillin, neomycin, and streptomycin). Cell culture medium was endotoxin free, as shown by the Limulus amebocyte lysate test. To study the effects of MS-8209 on TNF-␣ synthesis, monocyte-derived macrophages (MDM) were exposed to various concentrations of MS-8209 (0, 5, and 10 M) for 24 h, during which TNF-␣ was measured in cell supernatants by enzyme-linked immunosorbent assay (ELISA), and its mRNA was quantified by reverse transcriptase PCR (RT-PCR) (1). To investigate the effects of MS-8209 on HIV replication, 1 million MDM were infected in vitro with 10,000 50% tissue culture infective doses of either the reference macrophage-tropic HIV-1/Ba-L strain or the primary HIV-1-DAS isolate. TNF-␣ and viral replication were measured throughout the culture in cell supernatants by ELISA and by dosing RT activity, respectively, as previously described (7). In these last experiments, cell culture medium and MS-8209 (0, 1, 5, and 10 M) were renewed twice a week.MS-8209 dose dependently increased TNF-␣ synthesis and TNF-␣ mRNA expression during the first 24 h of MDM treat- RNAs were quantified using a noncompetitive RT-PCR (1), and the cytokine was detected in cell culture su...

Antibody Array-Generated Profiles of Cytokine Release from THP-1 Leukemic Monocytes Exposed to Different Amphotericin B Formulations

“…In fact, in a study by Prentice et al (15) there was a statistically higher success rate for resolution of fever with L-AMB treatment than with FZ treatment. Although the use of AMB derivatives has also been evaluated as a therapeutic approach to human immunodeficiency virus infection, the use of AMB formulations that induce TNF-␣ could increase the rate of human immunodeficiency virus replication, as reported previously (4). However, in the nonimmunocompromised patient, as long as nephrotoxicity can be tolerated, a systemic fungal infection might be better treated with FZ or ABCD because the recruitment of neutrophils and monocytes may be enhanced by the expression of proinflammatory cytokines.…”

Reduction of NO Synthase Expression and Tumor Necrosis Factor Alpha Production in Macrophages by Amphotericin B Lipid Carriers

“…The production of proinflammatory cytokines such as IL-1 (8,34), 32), and TNF-␣ has been proposed as a cause of some side effects of AMB, while treatments which reduce this production, namely, the use of antibodies (32) or dexamethasone (9) and pentoxilline (11), can reduce this toxicity. In particular, TNF-␣ production induced by AMB and its derivatives in human immunodeficiency virus-infected macrophages can stimulate virus replication (10). Therefore, AMB formulations which minimize TNF-␣ production would be advantageous for therapy of infections in AIDS patients.…”

Involvement of Macrophages in the Pathogenesis of Transmissible Spongiform Encephalopathies

“…activating macrophages in this inflammatory-like model (Manuelidis et al, 1997;, similarly to its effects on HIV-1 replication in monocyte-derived macrophages (Duval et al, 1997;Clayette et al, 1999).…”