A lymph node slice culture model to characterize T cell activation dynamics and anti-viral responses in human tissue
Year: 2023
Type: conference-abstract
Abstract: T cell activation in the lymph node (LN) is critical to initiating adaptive immune responses to vaccination, tumors, and infection. We previously demonstrated that T cells from human tissue sites (blood, LNs, bone marrow, lung) undergo conserved cell state transitions during activation. However, these studies were performed in suspension culture from dissociated tissue, depleting the spatial and signaling contributions of the tissue microenvironment. To address this, we developed techniques for culture and single-cell analysis of human LN slices, which preserve tissue architecture, local signaling, and physiological cell frequencies. Single-cell RNA-seq (scRNA-seq) of matched LN suspension and slice cultures stimulated with anti-CD3/28 antibody tetramers revealed distinct activation patterns. While anti-CD3/28 stimulation is commonly used to study T cell activation, this reagent circumvents antigen presentation and may not fully recapitulate antigen-mediated signaling. Conversely, antigen-dependent stimulation with superantigen mimics require MHCII/TCR binding and may benefit from the intact microenvironment in slice culture. We discovered a distinct metabolic intermediate through scRNA-seq analysis of superantigen-stimulated LN suspension and slice cultures that is not observed with anti-CD3/28 stimulation. Overall, these studies demonstrate the utility of analyzing T cell responses in intact tissue and comparative analysis of distinct mechanisms of activation. 1T32GM145440-01 (MSTP Training Grant) U19AI128949 (NIAID)
Source: The Journal of Immunology
Institution: Columbia University Irving Medical Center
Language: English
Topic: T-cell and B-cell Immunology
Subfield: Immunology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: bronze
OpenAlex ID: W4385685840