Dissecting the heterogeneity of DENV vaccine-elicited cellular immunity using single-cell RNA sequencing and cellular metabolic profiling

Abstract

Generating effective and durable T cell immunity is a critical prerequisite for vaccination against dengue virus (DENV) and other viral diseases. Understanding the precise molecular mechanisms of vaccine-elicited T cell immunity remains a critical knowledge gap in vaccinology. In this study, we utilized single-cell RNA sequencing (scRNAseq) and TCR clonotype analysis to demonstrate that a unique transcriptional signature is present in acutely-activated and clonally-expanded T cells that become committed to the memory repertoire. This effector-associated transcriptional signature is dominated by a unique metabolic transcriptional program. Based on this transcriptional signature, we were able to define a set of functional markers that identify the most potent and durable vaccine-reactive memory precursor CD8+ T cells. The transferrin receptor (CD71) other important transporters of amino acids, and direct measurements of glucose and fatty acid uptake, were further validated as early markers of durable T cell memory using conventional flow cytometry. These data suggest that generating durable T cell immunity is a process that is determined by metabolic programming and metabolite availability during the acute phase of the immune response. This study illustrates the power of scRNAseq as an analytical tool to assess the molecular mechanisms of host control and vaccine modality in determining the magnitude, diversity, and persistence of vaccine-elicited cell-mediated immunity.