Tuberculosis remains an old contagious disease that still has an impact on humanity. Despite the development of new diagnostic tools and drugs, the annual tuberculosis death rate remains high. It is known that, statistically, around 5–10% of people infected by MTB will actually develop active TB, due to the balance of immune system between host and the mycobacterium. Nucleotide-binding oligomerization domain-containing protein 2 (NOD2) is part of the Pattern Recognition Receptors (PRR) which play the first role in recognizing invasion from MTB through NLRs (NOD-like Receptors). Specific role of NOD2 is the response to a fragment of the bacterial cell wall, muramyl dipeptide (MDP), and in turn activated NF-κB to release pro inflammatory cytokine e.g TNF-α. Several studies have reported on the activity of this NOD2 gene, but there are still not many that link it to the effect of treatment on pulmonary TB. This study was conducted to determine the expression of the NOD2 gene and the levels of the pro-inflammatory cytokine TNF-α in pulmonary TB patients before and after 2 months of treatment. This study was a cohort study that followed patients for two months of treatment. This study included 36 patients diagnosed as new cases of pulmonary tuberculosis, all of whom were over the age of 18 and willing to sign informed consent. TNF- levels were investigated using the ELISA method, and NOD2 gene expression was investigated using the PCR method. There was a tendency to decrease NOD2 gene expression and TNF-α levels after 2 months of treatment, although it was not statistically significant. NOD2 gene expression was found to be significantly higher after 2 months of treatment with TNF-α (p = 0.029). Changes in TNF-cytokine serum levels vary in this study, but tend to decrease after 2 months of ATT. This study also shows a decrease in NOD2 gene expression, and we discovered the possibility of TNF- against NOD2 gene regulation after 2 months of ATT.