Alteration in HLA-DR, CD64, CD206 and CD163 marker expression in transduced macrophages with MARCH-1
نویسندگان: Ali Reza Andalib ©, Zivar Zangeneh, Gholamreza Khamisipour
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Background and Aim
Background: studies have shown that macrophage polarization is a therapeutic target for controlling immune responses especially in chronic inflammatory conditions. MARCH family protein (Membrane-Associated RING-CH-type finger) is known to inhibit dendritic cell maturation and antigen delivery to T lymphocytes. We transduce macrophages with MARCH-1 gene and assay their immunophenotypes to consider the macrophage plasticity to M2. Those could have beneficial for preventing covid-19 cytokine storm progression.
Methods: Plasmids pCDH-1 MARCH-1- GFP, psPAX2.221 and PMD2.G were used for transferring MARCH-1 into the macrophage. Macrophages derived from THP-1 cells and human monocytes were transduced with viral lentivirus produced from HEK-293T cells. transduced cells were stained with Anti-CD206-PE, Anti-CD163-PE, Anti-HLA-DR-PE, Anti-CD64-PE, Anti-CD14-APC markers for flow cytometry consideration. Mean fluorescence intensity (MFI) were assessed for each marker separately.
Results: MARCH-1GFP+ expressing macrophage population increase by 69.3% compared to non-transduced population. The mean fluorescent intensity (MFI) of HLA-DR expression in MARCH-1+ macrophage population reduced from 33135 to 13419 arbitrary unit (P=0.008) and CD64 decreased from 36695 to 28485 arbitrary unit as well (P=0.015). However, CD206 expression in transduced macrophages increased from 100.613.7 to 16822 arbitrary unit (P=0.008). The MFI for CD163 expression in transduced macrophages was 21555 in compared with non-transduced with 199.885 arbitrary unit (P=0.54).
Conclusion: This study has offered a detailed characterization of the polarizing effect of MARCH-1 on human monocyte-derived macrophages. Our data suggest that an increased expression of MARCH-1 in monocytes/ macrophages could be responsible for macrophage polarization to M2 macrophages. Thus targeting MARCH-1 in monocytes/macrophages could represent a new therapeutic strategy in inflammatory or anti inflammatory conditions such as covid-19 infection.
MARCH-1, HLA-DR, M2 macrophages, CD64
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