Minimizing Macrophage Extracellular Trap Formation in the Development of Pulmonary Tuberculosis

Abstract Background: Due to the rising frequency of antimicrobial treatment resistance and issues in vaccine development. To eradicate the microbiological infection with Mycobacterium tuberculosis can cause macrophages to release proinflammatory cytokines such as IL-1β, IFN- γ , and TNF- α . Objectives: The aim of the current study was to investigate the role of the TLR2 receptor in positively regulating IFN- γ and TNF- α production during M. bovis infected macrophages in vivo using human monocytic cell lines THP-1 and developed a new treatment to eliminate TB using TLR2 short interfering ribonucleic acid knockdown. Materials and Methods: The human monocytic cell line THP-1 cells were grown in the culture media that were prepared according to the manufacturer’s instructions. Short interfering ribonucleic acid (siRNA) was transfected into cells using (lipofectamine) DNA-fectin TM Plus transfection reagent. Total RNA from the macrophages was extracted, and the total RNA was measured using a NanoDrop spectrophotometer. The mRNA in the total RNA was reverse transcribed to complementary DNA using a cDNA synthesis Kit. Results: Showed that the relative expression of both TLR2 and IFN- γ was scaled down to 0.001% in TLR2 siRNA knockdown (TLR2-KD) samples compared to (TLR2-C) control without BCG vaccine, due to the TLR2 siRNA knockdown, whereas the TNF α relative expression was stabilized in both (TLR2-KD) and (TLR2-C) samples without BCG vaccine. Conclusion: TLR2 expression in THP-1 cells was consistent with our goal of reducing TLR2 expression. Our results showed a decrease in gene expression of TNF- α and IFN- γ in the TLR2 siRNA knockdown samples pluse BCG challenged after only 4h, which, in turn, reduced the synergistic effect of these genes (TNF- α and IFN- γ ) in granuloma formation during pulmonary tuberculosis.