Characterization of the anti-HIV effects of native lactoferrin and other milk proteins and protein-derived peptides.
In a search for natural proteins with anti-HIV activity, we screened a large set of purified proteins from bovine milk and peptide fragments thereof. Because several charged proteins and peptides are known to inhibit the process of virus entry, we selected proteins with an unusual charge composition or hydrophobicity profile. In contrast with some chemically modified (strongly negative) milk proteins, unmodified alpha(s2)-, beta- and kappa-casein, as well as several negatively and positively charged fragments thereof, did not show significant inhibition of virus replication. In fact, HIV-1 replication was elevated in the presence of beta-casein or amphiphilic fragments thereof. Bovine lactoferrin (bLF), a milk protein of 80 kDa, showed considerable inhibitory activity against HIV-1 with an IC50 of 0.4 microM. Modest inhibition was obtained with lactoferricin, a highly positively charged loop domain of bLF, indicating that other domains within the native bLF protein may also be required for inhibition. bLF blocked HIV-1 variants that use either the CXCR4 or the CCR5 coreceptor. In order to obtain further insight into the mechanism of action of this antiviral protein, we selected a bLF-resistant HIV-1 variant. The bLF-resistance phenotype is mediated by the viral envelope protein, which contains two interesting mutations that have previously been associated with an altered virus-host interaction and a modified receptor-coreceptor interaction. These results demonstrate that bLF targets the HIV-1 entry process.
Berkhout
Ben
B
Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands. b.berkhout@amc.uva.nl
van Wamel
Jeroen L B
JL
Beljaars
Leonie
L
Meijer
Dirk K F
DK
Visser
Servaas
S
Floris
René
R
D000998:Antiviral Agents / Q000494:pharmacology*; D065100:CCR5 Receptor Antagonists; D002460:Cell Line; D005814:Genes, Viral; D015497:HIV-1 / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D007781:Lactoferrin / Q000494:pharmacology*; D008894:Milk Proteins / Q000494:pharmacology*; D010446:Peptide Fragments / Q000494:pharmacology; D019718:Receptors, CXCR4 / Q000037:antagonists & inhibitors; D014759:Viral Envelope Proteins; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D065100:CCR5 Receptor Antagonists; D008894:Milk Proteins; D010446:Peptide Fragments; D019718:Receptors, CXCR4; D014759:Viral Envelope Proteins; D007781:Lactoferrin