![]() |
![]() |
||||||||||||
|
02202 STRUCTURAL STUDIES OF APTAMERS THAT NEUTRALIZE R5 SRTAIN OF HIV-1 Dey, Antu*, ** Human Immunodeficiency Virus 1(HIV-1) has evolved various strategies in response to the current anti-retroviral drugs and the selection pressure of humoral and cellular immunity. In particular, the R5 strains of HIV that is central to Acquired Immune Deficiency Syndrome (AIDS) pathogenesis are resistant to neutralization by antibodies. The recessed nature of the binding pockets on the surface glycoprotein (gp120), masking by hypervariable loops, conformational changes in the core gp120 molecule and a glycan shield mechanism of neutralization escape allow HIV-1 persistence in the face of an evolving antibody repertoire. We hypothesized that RNA aptamers, in contrast to antibodies, by virtue of their small size and slow dissociation rates, might be able to bind to the occluded neutralization sites on the gp120. Accordingly, using Systematic Evolution of Ligands by EXponential Enrichment (SELEX), we have isolated specific 2Fluoro-pyrimidine RNA aptamers that bind to monomeric HIV-1BaL gp120 with affinities in the order of 10-9 M. The aptamers not only neutralize HIV-1BaL in human Peripheral Blood Mononuclear Cells (PBMCs) by 1,000- to 100,000-fold but also neutralize clinical isolates from multiple clades efficiently. Secondary structure analysis, in conjunction with ribonuclease footprinting studies, revealed the probable region essential for gp120-binding. Truncating the aptamer to this smaller region retained its binding to monomeric gp120. The binding affinity and the neutralization potency of the truncated aptamer and the parental aptamer have been found to be similar. Further investigation of the structure of the truncated aptamer might lead to alternative anti-HIV-1 drugs and help us understand the molecular interaction between the viral gp120 and host cellular receptors that facilitate HIV-1 entry. Mr. Antu Dey |
|||||||||||||