Does the production of NF-ĸβ, IRF3, and IFN-β after LOS/LPS exposures in naive-HIV dendritic cells depend on TLR4-MD2 receptor pathway? In vitro and in silico study

• Published in: Journal of biotech research 2024-01, Vol.16, p.64-76
• Main Authors: Budiarti, Niniek, Kalim, Handono, Waafi, Affa Kiysa, Wulandari, Dewi Sri, Wahono, Cesarius Singgih, Manugan, Reizal Audi, Pradipto, Wiryawan, Jayanto, Galih Dwi, Santosaningsih, Dewi, Fitriana, Nur, Winaris, Nuning, Pawestri, Aulia Rahmi, Fitri, Loeki Enggar
• Format: Article
• Language: English
• Subjects: Affinity; Amino acids; Antigens; Bacterial infections; Binding; Cell culture; Cytokines; Dendritic cells; Drug development; E coli; Enzyme-linked immunosorbent assay; Exposure; Gonorrhea; HIV; Human immunodeficiency virus; Immune response; Immune system; Infections; Inflammation; Inflammatory response; Interferon; Interferon regulatory factor; Interferon regulatory factor 3; Ligands; Lipids; Lipooligosaccharides; Lipopolysaccharides; Molecular docking; Monocytes; Pathogenesis; Potassium; Proteins; Receptors; Simulation; TLR4 protein; Toll-like receptors; β-Interferon
• ISSN: 1944-3285

Lipopolysaccharide (LPS) and lipooligosaccharide (LOS) are inflammatory response inducers triggering downstream inflammatory signals by binding to the TLR4-MD2 complex. LPS/LOS exposure through TLR4 receptor on dendritic cells (DC) causes an increase in interferon (IFN)-p expression via an increase in interferon regulatory factor (IRF)3 expression and a decrease in nuclear factor (NF)-Kp, which result in HIV replication inhibition. This study aimed to compare the binding affinity between Escherichia coli LPS and Neisseria gonorrhoeae LOS toward TLR4-MD2 in silico and the production of NF-Kp, IRF3, and IFN-p from naive-HIV monocyte-derived dendric cells (MDDCs) after LPS and LOS exposure in vitro, to find a deeper understanding of host immune response, disease pathogenesis, and trends of drug development in the future. Molecular docking and dynamics were analyzed using AutoDock Vina and YASARA, respectively. The naive-HIV DC culture was exposed to LPS at 50,100, and 200 ng/mL, or LOS at concentrations of 2.5, 5, and 10% for 24-h. Levels of NF-Kp, IRF3, and IFN- p were measured by Enzymelinked Immunosorbent Assay (ELISA). The results showed that LOS demonstrated higher binding affinity to TLR4MD2 complex than LPS, although the docking between LOS and TLR4-MD2 complex involved fewer amino acids. Inversely, LPS exposure significantly increased IRF3 and IFN-p production (P < 0.01). Production of IFN-p was significantly increased with higher doses of LOS (P < 0.01). Different doses of LPS induced significant differences in NF-Kp and IFN-p levels (P< 0.01 and P< 0.05, respectively). Despite LOS showing higher binding affinity to TLR4MD2 complex, LPS exposure induced higher production of IRF3 and IFN-p from MDDCs. These findings provided information for deeper apprehension of immune responses and disease pathogenesis during HIV co-infections.