Research objectives and content
The identity of the long-elusive accessory factors for cellular infection by HIV- 1 has recently been shown to be family of seven-pass membrane proteins that naturally function as cellular receptors for the chemotactic and immunomodulatory chemokine proteins. However, many remaining questions concerning their role in infection in vivo have been hampered by the lack of highly specific and sensitive reagents. We aim to develop such reagents and, in doing so, shall investigate their potential as therapeutic agents. More specifically:
1. To develop novel ligands for chemokine receptors based on the selective evolution of ligands by exponential enrichment (SELEX) method. 2. To use biotinylated synthetic peptides derived from the extracellular portions of CXCR4, CCR5, CCR2b and CCR3 as selective agents and 2'-NH2 modified RNA as the library in physiological saline.
3. To screen the artificial ligands (aptamers) thus generated for specificity vs. cross-reactivity; for their ability to block chemokine receptor function; for their ability to block HIV- 1 infection in various primary and established cell types; for their affinity and binding kinetics to chemokine receptor-derived peptide in vitro; for their utility as flow cytometric and cytological immunofluorescent reagents for the detection of chemokine receptor expression.
Training content (objective, benefit and expected impact)
Acquisition of a range of skills in the following, leading-edge areas: molecular biology, nucleic acid chemistry, computer modelling, cell biology, virology. Experience of working in a major international institution with English as the principal means of communication. Links with industry I industrial relevance (22)
The sponsoring laboratory has links with the European nucleic acid chemistry-based pharmaceutical company, VPI, concerning related work. With synthetic RNAs. Glaxo-Wellcome, plc, has offered to provide training and support for this project.