In my talk, I will introduce the integrated approach that we have implemented in my lab to better understand HIV-1 fusion and infection in primary cells. Mathematical modelling combined with time-resolved data from advanced imaging (single virus tracking and Real-Time BlaM) allowed us to characterize HIV-1 priming, entry and fusion in live cells. I will also introduce a technique we have recently implemented in collaboration with other researchers in the WTCHG/University of Oxford: integrated single-cell transcriptomics and imaging to understand latency in HIV-1 resting primary CD4+ T cells. Combining single cell RNAseq with the beta-lactamase assa (BlaM assay) we want to address latency and the importance of pre-integrated viral DNA in resting CD4+ T cells. The BlaM assay utilizes engineered viruses able to release viral particles labelled with a BlaM protease (Vpr-BlaM) that in turn cleave a substrate previously loaded in target cells changing the spectrum from green (~520nm) to blue (~447nm). These assays are performed on microfluidic devices primed with primary resting CD4+ T cells from healthy individuals, allowing to study fusion events in conjunction with single RNAseq. We aim to understand why there is heterogeneity in HIV-1 fusion despite cells being genetically identical and exposed to high multiplicities of infection, and ultimately characterize the factors implicated in this process.


Wednesday, January 20, 2016, 12:00. Seminar Room

Hosted by Prof. María García-Parajo