Mass Cytometry and Single‐Cell RNA‐seq Profiling of the Heterogeneity in Human Peripheral Blood Mononuclear Cells Interacting with Silver Nanoparticles

Abstract

Understanding the interactions between nanoparticles (NPs) and human immune cells is necessary for justifying their utilization in consumer products and biomedical applications. However, conventional assays may be insufficient in describing the complexity and heterogeneity of cell–NP interactions. Herein, mass cytometry and single‐cell RNA‐sequencing (scRNA‐seq) are complementarily used to investigate the heterogeneous interactions between silver nanoparticles (AgNPs) and primary immune cells. Mass cytometry reveals the heterogeneous biodistribution of the positively charged polyethylenimine‐coated AgNPs in various cell types and finds that monocytes and B cells have higher association with the AgNPs than other populations. scRNA‐seq data of these two cell types demonstrate that each type has distinct responses to AgNP treatment: NRF2‐mediated oxidative stress is confined to B cells, whereas monocytes show Fcγ‐mediated phagocytosis. Besides the between‐population heterogeneity, analysis of single‐cell dose–response relationships further reveals within‐population diversity for the B cells and naïve CD4+ T cells. Distinct subsets having different levels of cellular responses with respect to their cellular AgNP doses are found. This study demonstrates that the complementary use of mass cytometry and scRNA‐seq is helpful for gaining in‐depth knowledge on the heterogeneous interactions between immune cells and NPs and can be incorporated into future toxicity assessments of nanomaterials.

Mass Cytometry Exploration of Immunomodulatory Responses of Human Immune Cells Exposed to Silver Nanoparticles

“…Significant cellular associations of Ag NPs were observed in monocytes and dendritic cells (DC), which is in good agreement with the previous studies [32,33] and current understanding of the cellular association of NPs; since monocytes are prominent antigen-presenting cells (APCs), they have a natural tendency to ingest foreign particles via phagocytosis [57,58]. Previous studies [34,55] have suggested that antigen-presenting cells recognize nanoparticles as foreign antigens, engulf and process them, then present them to effector cells via the major histocompatibility complex, thereby priming the antigen-specific cellular immune response.…”

Recent Advances in Developmental Hematopoiesis: Diving Deeper With New Technologies

“…CyTOF was also instrumental in identifying a pro-inflammatory subset of macrophages that is involved in the development of HSCs ( 152 ). In addition, integrating CyTOF with scRNA-seq could provide additional discriminatory power for further sub-setting or functional analysis between distinct subsets of cells like HSCs and progenitors ( 153 ). Besides the precise immunophenotyping of cells in suspension, recent advances have enabled CyTOF of tissue sections or cells cultured on a slide.…”

Human Monocytes Plasticity in Neurodegeneration

“…Moreover, a further development of the technology is represented by RNAseq at the single cell level, which is the most recent major achievement in transcriptomics analysis. Single cell analysis promises to finally dissect the subset subdivisions within the immune cell subpopulations [28]. Blood monocyte scRNAseq indicated that classical and non-classical monocytes belong to two major transcriptionally defined clusters [29].…”