Mycobacterial infections remain leading causes of morbidity and mortality globally. The pro-inflammatory cytokine TNF is paramount to effective host immune responses against mycobacterial infections. Expression and release of TNF are tightly regulated, yet the molecular mechanisms that control the release of TNF by mycobacteria-infected host cells, in particular macrophages, remain to be defined. Rab proteins are small GTPases that direct the transport of intracellular membrane-enclosed vesicles and are important regulators of macrophage cytokine secretion. The Golgi-associated GTPases Rab6a and Rab6a’ are known to positively regulate the trafficking of TNF in LPS-activated macrophages. Unlike Rab6a and Rab6a’ which are ubiquitously expressed, the related family member, Rab6b, is highly expressed in human and mouse neuronal tissues. Functional studies in brain-derived cells have reported Rab6b contributions to retrograde vesicular transport and suggested a role for this protein in secretory trafficking for exocytosis. Whether Rab6b executes similar functions in the context of immune responses is unknown. Here we show that Rab6b is expressed by primary mouse macrophages and localises to the Golgi complex in a fashion similar to Rab6a. Infection with Mycobacterium bovis Bacille Calmette Guérin (BCG) led to dynamic changes in Rab6b expression in primary mouse macrophages in vitro as well as organs of infected mice in vivo. Rab6b facilitated TNF release by BCG-infected macrophages, in the absence of discernible impact on Tnf mRNA and intracellular TNF protein expression. Similarly, Rab6a promoted BCG-induced TNF release by macrophages without impacting TNF protein expression. Our observations identify a positive regulatory role for Rab6a and Rab6b in BCG-induced TNF secretion by macrophages, and positions both proteins among the molecular machinery that direct macrophage functions in the context of infection and inflammation.