Biology subjects

Lanier, L.

Publications and source records attributed to Lanier, L..

2 recordsLinked to original sources

bioRxiv2025

FcεR1γ-based Activating Chimeric Antigen Receptors enhanced Natural Killer cell function against HLA-E+ cells

Fc{varepsilon}R1{gamma} is a transmembrane adaptor protein regulating NKp30, NKp46, and CD16 expression and function in human NK cells. HLA-E expression by cancer cells suppresses NK cell function through interaction with NKG2A. Here, we engineered human NK cells to express a modified Fc{varepsilon}R1{gamma} with a 4-1BB co-stimulatory motif. Fc{varepsilon}R1{gamma}41BB expression led to upregulation of NKp30, NKp46, and CD16 expression and enhanced NKp30 and CD16-mediated target cell lysis. Against cells expressing HLA-E, Fc{varepsilon}R1{gamma}41BB improved NK cell lytic function relative to Fc{varepsilon}R1{gamma}. We then generated an CD19ScFv-CD8hinge-Fc{varepsilon}R1{gamma}41BB(TM-IC) CAR (FCRG10). Engineering human NK cells to express FCRG10 led to upregulation of NKp30 and CD16, and improved NK cell-mediated cytotoxicity against cells co-expressing B7H6, CD19, and HLA-E relative to NK cells expressing the conventional CD19CAR41BB-CD3{zeta} with a CD8 transmembrane domain. Our results indicate that modification of Fc{varepsilon}R1{gamma} enhanced NK cell lytic function mediated by several activating receptors and improved CAR NK cell response in conditions favoring NK cell suppression by HLA-E. Our work may contribute to NK cell-based cancer immunotherapy mediated by co-recognition of stress-induced ligands, antibody-coated cells, and tumor-associated antigens.

immunology↗

bioRxiv2019

A Modified Injector and Sample Acquisition Protocol Can Improve Data Quality and Reduce Inter-Instrument Variability of the Helios Mass Cytometer

Mass cytometry is a powerful tool for high dimensional single cell characterization. Since the introduction of the first commercial CyTOF mass cytometer by DVS Sciences in 2009, mass cytometry technology has matured and become more widely utilized, with sequential platform upgrades designed to address specific limitations and to expand the capabilities of the platform. Fluidigms 3rd-generation Helios mass cytometer introduced a number of upgrades over the previous CyTOF2. One of these new features is a modified narrow bore sample injector that generates smaller ion clouds, which is expected to improve sensitivity and throughput. However, following rigorous testing we find that the narrow-bore sample injector may have unintended negative consequences on data quality and result in lower median signal intensities and higher coefficients of variation in antibody expression. We describe an alternative Helios acquisition protocol using a wider bore injector, which largely mitigates these data quality issues. We directly compare these two protocols in a multi-site study of 10 Helios instruments across 7 institutions and show that the modified protocol improves data quality and reduces inter-instrument variability. These findings highlight and address an important source of technical variability in mass cytometry experiments that is of particular relevance in the setting of multi-center studies.

immunology↗