CD4-Specific Designed Ankyrin Repeat Proteins Are Novel Potent HIV Entry Inhibitors with Unique Characteristics

PLoS Pathogens

Stuart G. Turville¹  Meropi Aravantinou¹  Melissa Robbiani¹  Andreas Schweizer²  Alexandra Trkola²  Claudia R. Ruprecht²  Axel Mann²  Stéphanie Corthésy²  Livia Berlinger²  Marek Fischer²  Peter Rusert²  Patrick Amstutz³ 

[1] Center for Biomedical Research, Population Council, New York, New York, United States of America;Division of Infectious Diseases, University Hospital Zurich, Zurich, Switzerland;Molecular Partners AG, Zurich-Schlieren, Switzerland

关键词: HIV;    Macaque;    T cells;    Microbicides;    CD coreceptors;    Viral replication;    Enzyme-linked immunoassays;    T cell receptors;   

DOI  :  10.1371/journal.ppat.1000109

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Here, we describe the generation of a novel type of HIV entry inhibitor using the recently developed Designed Ankyrin Repeat Protein (DARPin) technology. DARPin proteins specific for human CD4 were selected from a DARPin DNA library using ribosome display. Selected pool members interacted specifically with CD4 and competed with gp120 for binding to CD4. DARPin proteins derived in the initial selection series inhibited HIV in a dose-dependent manner, but showed a relatively high variability in their capacity to block replication of patient isolates on primary CD4 T cells. In consequence, a second series of CD4-specific DARPins with improved affinity for CD4 was generated. These 2nd series DARPins potently inhibit infection of genetically divergent (subtype B and C) HIV isolates in the low nanomolar range, independent of coreceptor usage. Importantly, the actions of the CD4 binding DARPins were highly specific: no effect on cell viability or activation, CD4 memory cell function, or interference with CD4-independent virus entry was observed. These novel CD4 targeting molecules described here combine the unique characteristics of DARPins—high physical stability, specificity and low production costs—with the capacity to potently block HIV entry, rendering them promising candidates for microbicide development.

【 授权许可】

CC BY