CD16⁺ monocytes give rise to CD103⁺RALDH2⁺TCF4⁺ dendritic cells with unique transcriptional and immunological features

Classical CD16^- vs intermediate/nonclassical CD16⁺ monocytes differ in their homing potential and biological functions, but whether they differentiate into dendritic cells (DCs) with distinct contributions to immunity against bacterial/viral pathogens remains poorly investigated. Here, we employed a systems biology approach to identify clinically relevant differences between CD16⁺ and CD16^- monocyte-derived DCs (MDDCs). Although both CD16⁺ and CD16^- MDDCs acquire classical immature/mature DC markers in vitro, genomewide transcriptional profiling revealed unique molecular signatures for CD16⁺ MDDCs, including adhesion molecules (ITGAE/CD103), transcription factors (TCF7L2/TCF4), and enzymes (ALDH1A2/RALDH2), whereas CD16^- MDDCs exhibit a CDH1/E-cadherin1 phenotype. Of note, lipopolysaccharides (LPS) upregulated distinct transcripts in CD16⁺ (eg, CCL8, SIGLEC1, MIR4439, SCIN, interleukin [IL]-7R, PLTP, tumor necrosis factor [TNF]) and CD16^- MDDCs (eg, MMP10, MMP1, TGM2, IL-1A, TNFRSF11A, lysosomal-Associated membrane protein 1, MMP8). Also, unique sets of HIV-modulated genes were identified in the 2 subsets. Further gene set enrichment analysis identified canonical pathways that pointed to "inflammation" as the major feature of CD16⁺ MDDCs at immature stage and on LPS/HIV exposure. Finally, functional validations and meta-Analysis comparing the transcriptome of monocyte and MDDC subsets revealed that CD16⁺ vs CD16^- monocytes preserved their superior ability to produce TNF-α and CCL22, as well as other sets of transcripts (eg, TCF4), during differentiation into DC. These results provide evidence that monocyte subsets are transcriptionally imprinted/programmed with specific differentiation fates, with intermediate/nonclassical CD16⁺ monocytes being precursors for pro-inflammatory CD103⁺ RALDH2⁺TCF4⁺ DCs that may play key roles in mucosal immunity homeostasis/ pathogenesis. Thus, alterations in the CD16⁺/CD16^- monocyte ratios during pathological conditions may dramatically influence the quality of MDDC-mediated immunity.