Diabetes mellitus is an important risk factor for development of tuberculosis. Our previous study found that glibenclamide reduced neutrophil cytokine production, migration and killing in responses to bacterial infection. Glibenclamide is a widely used antidiabetic drug in low and middle-income countries where the incidence of tuberculosis is high. It is a broad-spectrum ATP-binding cassette transporter inhibitor and a K+ATP channel blocker which inhibits cryopyrin/Nalp3 inflammasome activation and proteolytic maturation of IL-1β and IL-18 by caspase-1 in neutrophils and macrophages in response to Burkholderia pseudomallei (melioidosis). In mice infected with M. tuberculosis (Mtb), the production of IL-1β is essential for host resistance through the enhancement of cyclooxygenase that limits excessive Type I interferon (IFN) production and fosters Mtb containment. Since melioidosis and tuberculosis share many immunological characteristics, we hypothesize that glibenclamide may also interfere with monocyte mediated immune responses against Mtb and alter the balance between protective IL-1β and immunosuppressive Type I interferon-mediated immunity. Here we demonstrate that purified human monocytes from diabetic individuals who had been treated with glibenclamide showed reduction of IL-1β and IL-8 secretion but revealed enhancement of bacterial growth when exposed to Mtb. Additionally, these responses also occurred when monocytes from non-diabetic control individuals were pre-treated with glibenclamide in vitro. Moreover, this pre-treatment enhanced IFN-α expression but not involved with prostaglandin level in response to Mtb infection. Taken together, our data show that glibenclamide might be responsible, at least in part, to the increased susceptibility of diabetic individuals to Mtb infection by reducing IL-1β production and killing of monocytes.