Kappa-opioid receptor agonist inhibition of HIV-1 envelope glycoprotein-mediated membrane fusion and CXCR4 expression on CD4(+) lymphocytes.
Our previous studies have shown that the suppressive effect of kappa-opioid receptor (KOR) ligand treatment on HIV-1(AT) (a T-tropic strain) expression in acutely infected CD4(+) lymphocytes is time-dependent. This finding implied that the inhibition observed following treatment with KOR agonists such as U50,488 (trans-3,4-dichloro-N-methyl-N[2-(1-pyrolidinyl)cyclohexyl]benzeneaceamide methanesulfonate) occurs at an early step in the viral replication cycle, perhaps as early as viral entry. In the present study, we examined the hypothesis that U50,488 treatment of CD4(+) lymphocytes inhibits HIV-1 envelope (Env) glycoprotein-mediated membrane fusion. We used a vaccinia virus-based assay to measure the effects of U50,488 treatment of CD4(+) lymphocytes on HIV-1 IIIB Env glycoprotein-mediated fusogenic activity, based on the cytoplasmic activation of a reporter gene. The results show that U50,488 inhibited Env-mediated cell fusion in a bell-shaped concentration-response manner with suppression ranging between 31 and 98% at concentrations of 10(-8) and 10(-10)M (N=9 experiments). U50,488 was also found to inhibit cell fusion when monitored in situ with 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) staining. Blockade of the inhibitory activity of U50,488 by the KOR antagonist nor-bialtorphimine (nor-BNI) suggested that U50,488 was acting via a KOR-related mechanism. Using flow cytometry, we demonstrated that the chemokine co-receptor CXCR4, but not CD4, is down-regulated as a consequence of KOR activation, with 44.2+/-3.5% suppression at 10(-10)M U50,488. These findings support the hypothesis that KOR-related activation of CD4(+) lymphocytes inhibits HIV-1 entry via down-regulation of CXCR4.
Lokensgard
James R
JR
Neuroimmunology Laboratory, Minneapolis Medical Research Foundation, 914 South 8th Street, Bldg. D-3, Minneapolis, MN 55404, USA. jlokensgard@mmrf.org
Gekker
Genya
G
Peterson
Phillip K
PK
D019900:3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer / Q000494:pharmacology*; D018712:Analgesics, Non-Narcotic / Q000494:pharmacology*; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects* / Q000378:metabolism; D005434:Flow Cytometry; D015870:Gene Expression / Q000187:drug effects*; D015699:HIV Envelope Protein gp120 / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D008561:Membrane Fusion / Q000187:drug effects*; D019718:Receptors, CXCR4 / Q000096:biosynthesis*; D017473:Receptors, Opioid, kappa / Q000819:agonists
D018712:Analgesics, Non-Narcotic; D015699:HIV Envelope Protein gp120; D019718:Receptors, CXCR4; D017473:Receptors, Opioid, kappa; D019900:3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer