ESAT-6 impedes IL-18 mediated phagosome lysosome fusion via microRNA-30a upon Calcimycin treatment in mycobacteria infected macrophages
Autor: | Assirbad Behura, Ramandeep Singh, Amit Mishra, Debraj Manna, Lincoln Naik, Ashish Kumar, Rohan Dhiman, Salina Patel, Abtar Mishra, Mousumi Das |
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Rok vydání: | 2021 |
Předmět: |
Immunology
Blotting Western Real-Time Polymerase Chain Reaction Cell Line Immune system Antigen Bacterial Proteins Phagosomes Immunology and Allergy Humans Tuberculosis Neutralizing antibody PI3K/AKT/mTOR pathway Calcimycin Pharmacology Phagosome-lysosome fusion Antigens Bacterial Microscopy Confocal biology Chemistry Interleukin-18 Transfection Mycobacterium tuberculosis Flow Cytometry Cell biology Anti-Bacterial Agents MicroRNAs biology.protein Lysosomes Intracellular |
Zdroj: | International immunopharmacology. 101 |
ISSN: | 1878-1705 |
Popis: | The weaponry possessed by Mycobacterium tuberculosis (M. tb) in the form of immunodominant antigens hijack the host immune system to give a survival advantage to this intracellular fiend, but the mechanism of this control is not entirely known. Since we have previously reported the mechanism of autophagy inhibition by early secreted antigenic target 6 kDa (ESAT-6) through microRNA (miR)-30a-3p in Calcimycin treated differentiated THP-1 (dTHP-1) cells, the present study was undertaken to deduce the effect of miR-30a on the immunomodulatory profile of ESAT-6 treated cells and the mechanism involved thereof, if any. Initially, the effect of recombinant ESAT-6 (rESAT-6) on the immunomodulatory profile in Calcimycin-treated phorbol 12-myristate 13-acetate (PMA) dTHP-1 cells was checked. Later, transfection studies using miR-30a-3p inhibitor or -5p mimic highlighted the contrary roles of different arms of the same miRNA in regulating IL-18 response by ESAT-6 in dTHP-1 cells after Calcimycin treatment. By using either IL-18 neutralizing antibody or inhibitors of phosphoinositide 3-kinase (PI3K)/NF-κB/phagosome-lysosome fusion in the miRNA-30a transfected background, IL-18 mediated signaling and intracellular killing of mycobacteria was reversed in the presence of ESAT-6. Overall, the results of this study conclusively prove the contrary roles of miR-30a-3p and miR-30a-5p in regulating IL-18 signaling by ESAT-6 in dTHP-1 cells upon Calcimycin treatment that affected phagosome-lysosome fusion and intracellular survival of mycobacteria. |
Databáze: | OpenAIRE |
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