Flow Cytometry for Investigating HIV Infection of Living Cells (360G-Wellcome-208354_Z_17_Z)

Funding is requested to purchase a Becton Dickinson FACSMelody Benchtop Cell Sorter System that will be dedicated to analysing and/or purifying living cells either obtained from HIV-1 infected patients, or purposefully infected with HIV-1 in culture. The sorter will be located in the KCL containment level 3 (CL3) laboratories at Guy’s Hospital. Flow cytometry is a recognised strength at KCL, but none of our facilities offer CL3 capability, thus prohibiting any flow cytometry-based analysis or purification of living cells infected with HIV-1, or experimental workflow that requires downstream steps involving living cells. The acquisition of the FACSMelody will be transformative for our HIV/AIDS research community, and for our collaborators, by enabling the pursuit of new projects as well as the application of additional technologies such as single cell cloning and analytics to existing projects. Our work will address key areas relevant to understanding HIV-1 immunopathogenesis, including: HIV-1’s ability to manipulate normal cell function, the role(s) of cell-encoded factors in productive HIV-1 replication, anti-viral innate immunity and mechanisms of viral evasion, the molecular basis for HIV-1 persistence/ latency and its reversal, adaptive T and B cell immune responses in infants and adults, and mechanisms of HIV-1 induced immune cell dysfunction.