The study found that when human immune cells are treated with active vitamin D, they become better at fighting the tuberculosis bacteria. These vitamin‑D‑treated cells make a lot of the antimicrobial peptide LL‑37 and keep the bacteria from growing, while avoiding the production of an immunosuppressive enzyme (IDO). This suggests vitamin D can boost a natural defense mechanism in the body.

<b>Background:</b> Understanding macrophage behavior is key to decipher <i>Mycobacterium tuberculosis</i> (Mtb) pathogenesis. We studied the phenotype and ability of human monocyte-derived cells polarized with active vitamin D [1,25(OH)₂D₃] to control intracellular Mtb infection compared with polarization of conventional subsets, classical M1 or alternative M2. <b>Methods:</b> Human blood-derived monocytes were treated with active vitamin D or different cytokines to obtain 1,25(OH)₂D₃-polarized as well as M1- and M2-like cells or fully polarized M1 and M2 subsets. We used an <i>in vitro</i> macrophage Mtb infection model to assess both phenotype and functional markers i.e., inhibitory and scavenger receptors, costimulatory molecules, cytokines, chemokines, and effector molecules using flow cytometry and quantitative mRNA analysis. Intracellular uptake of bacilli and Mtb growth was monitored using flow cytometry and colony forming units. <b>Results:</b> Uninfected M1 subsets typically expressed higher levels of CCR7, TLR2, and CD86, while M2 subsets expressed higher CD163, CD200R, and CD206. Most of the investigated markers were up-regulated in all subsets after Mtb infection, generating a mixed M1/M2 phenotype, while the expression of CD206, HLADR, and CD80 was specifically up-regulated (<i>P</i> &lt; 0.05) on 1,25(OH)₂D₃-polarized macrophages. Consistent with the pro-inflammatory features of M1 cells, Mtb uptake and intracellular Mtb growth was significantly (<i>P</i> &lt; 0.01-0.001 and <i>P</i> &lt; 0.05-0.01) lower in the M1 (19.3%) compared with the M2 (82.7%) subsets 4 h post-infection. However, infectivity rapidly and gradually increased in M1 cells at 24-72 h. 1,25(OH)₂D₃-polarized monocyte-derived cells was the most potent subset to inhibit Mtb growth at both 4 and 72 h (<i>P</i> &lt; 0.05-0.01) post-Mtb infection. This ability was associated with high mRNA levels of pro-inflammatory cytokines and the antimicrobial peptide LL-37 but also anti-inflammatory IL-10, while expression of the immunosuppressive enzyme IDO (indoleamine 2,3-dioxygenase) remained low in Mtb-infected 1,25(OH)₂D₃-polarized cells compared with the other subsets. <b>Conclusions:</b> Mtb infection promoted a mixed M1/M2 macrophage activation, and 1,25(OH)₂D₃-polarized monocyte-derived cells expressing LL-37 but not IDO, were most effective to control intracellular Mtb growth. Macrophage polarization in the presence of vitamin D may provide the capacity to mount an antimicrobial response against Mtb and simultaneously prevent expression of inhibitory molecules that could accelerate local immunosuppression in the microenvironment of infected tissue.