Haeseleer, F. | bioRxiv Science

bioRxiv2024

A suite of enhancer AAVs and transgenic mouse lines for genetic access to cortical cell types

The mammalian cortex is comprised of cells classified into types according to shared properties. Defining the contribution of each cell type to the processes guided by the cortex is essential for understanding its function in health and disease. We used transcriptomic and epigenomic cortical cell type taxonomies from mouse and human to define marker genes and putative enhancers and created a large toolkit of transgenic lines and enhancer AAVs for selective targeting of cortical cell populations. We report evaluation of fifteen new transgenic driver lines, two new reporter lines, and >800 different enhancer AAVs covering most subclasses of cortical cells. The tools reported here as well as the scaled process of tool creation and modification enable diverse experimental strategies towards understanding mammalian cortex and brain function.

molecular biology↗

bioRxiv2022

Novel engineered chimeric engulfment receptors trigger T-cell effector functions against SIV infected CD4+ T cells

Adoptive therapy with genetically engineered T cells offers potential for infectious disease treatment in immunocompromised persons. HIV/simian immunodeficiency virus (SIV) infected cells express phosphatidylserine (PS) early post-infection. We tested whether chimeric engulfment receptor (CER) T cells designed to recognize PS-expressing cells could eliminate SIV infected cells. Lentiviral CER constructs comprised of the extracellular domain of T-cell immunoglobulin and mucin domain containing 4 (TIM-4), the PS receptor, and engulfment signaling domains were transduced into primary rhesus macaque (RM) T cells. We measured PS binding and T-cell engulfment of RM CD4+ T cells infected with SIV expressing GFP. As chimeric antigen receptor (CAR) T cells induce PS and subsequent TIM-4 binding, we evaluated in vitro killing of CAR and CER T-cell combinations. We found that recombinant TIM-4 bound to SIV infected cells. In vitro, CER CD4+ T cells effectively killed SIV infected cells, which was dependent on TIM-4 binding to PS. Enhanced killing of SIV infected CD4+ T cells by CER and CAR T-cell combinations was observed. This installation of innate immune functions into T cells presents an opportunity to enhance elimination of SIV infected cells and offers potential to augment functional cure of SIV/HIV infection.

immunology↗