Mycobacterium tuberculosis (Mtb), the etiological agent of tuberculosis, caused an estimated 1.3 million deaths in 2015. Mtb is a resilient organism, persisting through long courses of antibiotics and years of dormancy within the host. Rv2626c protein is known as a Hypoxia-Related Protein-1 (Hrp-1), because the association with the DsoR/DosS regulon. Rv2626c expression of the gene is an important landmark for Mtb to enter dormancy and have been proposed to play a role in evasion of the host immune response since their expression levels differ under specific environmental conditions. Rv2626c antigens can stimulate the body to produce higher levels of antibodies. Therefore, the recombinant antigens Rv2626c have potential to be used as diagnostic markers for TB and need to evaluate with other antigens for differential diagnosis of TB. However, the roles of Rv2626c in toll-like receptor (TLR)s signaling remains largely unknown. Here we show that TLR ligands-induced innate immune responses and NLRP3 inflammasomes activation were regulated by Rv2626c in murine macrophages. Thus, these results provide novel insight into the crucial role of Mtb Rv2626c in TLR signaling-mediated innate immune responses.