Induced overexpression of MARCH-۱ in human macrophages altered to M۲ phenotype for suppressing inflammation process

Objective(s): The M۱ macrophage is characterized by enhanced pro-inflammatory cytokines production, whereas macrophage (M۲) has anti-inflammatory features. Macrophage polarization as a therapeutic target for controlling immune responses could be performed by gene transduction to control the regulation of exaggerated innate/adaptive immune responses. Materials and Methods: Macrophages were prepared from THP-۱ cell line and human monocytes that were transduced with (Membrane-Associated RING-CH-type finger) MARCH-۱ viral lentivector produced in HEK-۲۹۳T cells. RT-PCR and Western blotting confirmed MARCH-۱ gene transduction. Cytokine production, CD markers assay, macrophage phagocytosis potential activity and mixed leukocyte reaction (MLR) with CFSE were performed for M۱/M۲ plasticity.Results: The mean fluorescent intensity of HLA-DR and CD۶۴ expression reduced in MARCH-۱+ transduced macrophage population. However, CD۲۰۶ and CD۱۶۳ expression increased in these macrophages. The concentrations of IL-۶, TNF-α and iNOS were decreased in MARCH-۱ transduced cells, and TGF-β production showed an augmentation in concentration. Western blotting and real-time PCR measurement confirmed that the expression levels of MARCH-۱ protein and arginase-۱ enzyme were increased in transduced macrophages.Conclusion: The anti-inflammatory features of MARCH-۱ revealed the reduced levels of pro-inflammatory factors and maintained M۲ phenotype characterized by high levels of scavenger receptors. Therefore, targeting MARCH-۱ in monocytes/macrophages could represent a new autologous cell-based therapies strategy for inflammatory conditions.

کلیدواژه ها:

INOS ، Macrophage ، MARCH-۱ ، Polarization ، TGF-beta