Macrophages are important immune sentinels that detect and clear pathogens and initiate inflammatory responses through the activation of surface receptors, including Toll-like receptors (TLRs). Activated TLRs employ complex cellular trafficking and signalling pathways to initiate transcription for inflammatory cytokine programs. Small GTPases of the Rab family regulate trafficking and signalling throughout cells. We have previously shown that Rab8a regulates Akt/mTOR signalling downstream of multiple TLRs by recruiting the effector PI3Kg, but the guanine nucleotide exchange factor (GEF) canonically required for Rab8a activation in these pathways is not known. Two known Rab8 specific GEFs, Rabin8 and GRAB, were shown to be expressed in LPS-activated macrophages. Both Rabin8 and GRAB demonstrate LPS-inducible binding to Rab8 and are localized on cell surface ruffles and macropinosomes where they coincide with Rab8a localisation. CRISPR-Cas9 knock-out (KO) cell lines were created for gene deletion of Rabin8 or GRAB and for double KOs. Evidence from these KOs showed that Rabin8 and GRAB both contribute to and are jointly sufficient for Rab8a activation in response to LPS. However, interestingly, GRAB and Rabin8 differentially affect TLR4 signalling events such as Akt phosphorylation and the activation of mTOR associated substrates PRAS40 and p70. We propose that Rabin8 and GRAB activate Rab8a for different roles downstream of TLR activation to guide pathogen-directed responses and inflammation.  The Rab8a GEFs thus contribute to the molecular cascades triggered by TLRs that help bias signalling, cytokine outputs and macrophage inflammatory programs.