Cytokine/Chemokine Release Patterns and Transcriptomic Profiles of LPS/IFNγ-Activated Human Macrophages Differentiated with Heat-Killed Mycobacterium obuense, M-CSF, or GM-CSF

Macrophages (Mφs) are instrumental regulators of the immune response whereby they acquire diverse functional phenotypes following their exposure to microenvironmental cues that govern differentiation from monocytes and activation. The complexity diversity mycobacterial cell wall have empowered mycobacteria with potent immunomodulatory capacities. A heat-killed (HK) whole-cell preparation Mycobacterium obuense (M. obuense) has shown promise as an adjunctive immunotherapeutic agent for treatment cancer. Moreover, HK M. been trigger human into a monocyte-derived macrophage (MDM) type named Mob-MDM. However, transcriptomic profile properties Mob-MDMs remain undefined during activation state. Here, we characterized cytokine/chemokine release patterns profiles lipopolysaccharide (LPS)/interferon γ (IFNγ)-activated MDMs were differentiated (Mob-MDM(LPS/IFNγ)), colony-stimulating factor M-MDM(LPS/IFNγ)), or granulocyte/macrophage (GM-MDM(LPS/IFNγ)). Mob-MDM(LPS/IFNγ) demonstrated unique pattern (interleukin (IL)-10low, IL-12/23p40low, IL-23p19/p40low, chemokine (C-x-C) motif ligand (CXCL)9low) was distinct those M-MDM(LPS/IFNγ) GM-MDM(LPS/IFNγ). Furthermore, maintained IL-10 production at significantly higher levels compared GM-MDM(LPS/IFNγ) despite being activated M1-Mφ-activating stimuli. Comparative RNA sequencing analysis pointed transcriptome relative both comprised 417 transcripts. Functional gene-set enrichment revealed significant overrepresentation signaling pathways biological processes uniquely related Mob-MDM(LPS/IFNγ). Our findings lay foundation potential integration in specific cell-based modalities such adoptive transfer Mφs (Mob-MDM(LPS/IFNγ)) cancer treatment.