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    Peripheral monocytes in humans are conventionally divided into classical (CL, CD14++CD16-), intermediate (INT, CD14++CD16+) and non-classical (NC, CD14dim/-CD16++) cells, based on their expression levels of CD14 and CD16. A major fraction of the NC-monocytes has been shown to express the 6-sulfo LacNAc (slan) antigen, but whether these slan+/NC-monocytes represent the prototypical non-classical monocytes or whether they are simply a sub-fraction with identical features as the remainder of NC monocytes is still unclear. We analyzed transcriptome (by bulk and single cell RNA-seq), proteome, cell surface markers and production of discrete cytokines by peripheral slan+/NC- and slan-/NC-monocytes, in comparison to total NC-, CL- and INT- monocytes. By bulk RNA-seq and proteomic analysis, we found that slan+/NC-monocytes express higher levels of genes and proteins specific of NC-monocytes than slan-/NC-monocytes do. Unsupervised clustering of scRNA-seq data generated one cluster of NC- and one of INT-monocytes, where all slan+/NC-monocytes were allocated to the NC-monocyte cluster, while slan-/NC-monocytes were found, in part (13.4%), within the INT-monocyte cluster. In addition, total NC- and slan-/NC-monocytes, but not slan+/NC-monocytes, were found by both bulk RNA-seq and scRNA-seq to contain a small percentage of natural killer cells. In addition to comparatively characterize total NC-, slan-/NC- and slan+/NC-monocyte transcriptomes and proteomes, our data prove that slan+/NC-, but not slan-/NC-, monocytes are more representative of prototypical NC-monocytes. Copyright © 2023 Tamassia, Bianchetto-Aguilera, Gasperini, Grimaldi, Montaldo, Calzetti, Gardiman, Signoretto, Castellucci, Barnaba, Tripodi and Cassatella.

    Citation

    Nicola Tamassia, Francisco Bianchetto-Aguilera, Sara Gasperini, Alessio Grimaldi, Claudia Montaldo, Federica Calzetti, Elisa Gardiman, Ilaria Signoretto, Monica Castellucci, Vincenzo Barnaba, Marco Tripodi, Marco Antonio Cassatella. The slan antigen identifies the prototypical non-classical CD16+-monocytes in human blood. Frontiers in immunology. 2023;14:1287656

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    PMID: 37965335

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