T cell populations are negatively correlated with natural killer and macrophage cell populations in aspirate samples of peripheral lymphadenopathies.

We employed single-cell RNA sequencing (scRNA-seq) of fine needle aspirates (FNAs) to describe the cells and communication networks characterizing granulomatous lymph nodes of TB patients. We uniformly identified several cell types known to characterize granulomas. Overall, we found the T cell cluster to be the most abundant. Other cell clusters that were uniformly detected, but that varied in abundance amongst the individual patient samples, were the B cell, plasma cell and macrophage/dendritic and NK cell clusters. When we combined all our scRNA-seq data from our current 19 patients, we distinguished T, B, macrophage, dendritic and plasma cell subclusters. The sizes of these subclusters also varied dramatically amongst the individual patients. In comparing FNA composition we noted trends in which T cell populations were negatively correlated with NK cell populations and with macrophage/dendritic cell populations. In addition, we discovered that the scRNA-seq pipeline detects Mtb RNA transcripts and associates them with their host cell's transcriptome, thus identifying individual infected cells. The number of infected cells also varies in abundance amongst the patient samples. CellChat analysis identified predominating signaling pathways amongst the cells comprising the various granulomatous lymph nodes, identifying several pathways involved in immune cell maturation, migration and adhesion.