Despite representing a very important class of virulence proteins, the role of lipoproteins in the pathogenesis ofremains elusive. In this study, we investigated the role of putative lipoprotein LprE in the subversion of host immune responses using theCDC1551 LprE (LprE) mutant (∆LprE). We show that deletion of LprEresults in reduction ofvirulence in human and mouse macrophages due to upregulation of vitamin D3-responsive cathelicidin expression through the TLR2-dependent p38-MAPK-CYP27B1-VDR signaling pathway. Conversely, episomal expression of LprEinimproved bacterial survival. Infection in siTLR2-treated ormacrophages reduced the survival of LprEexpressingandbecause of a surge in the expression of cathelicidin. Infection with the LprEmutant also led to accumulation of autophagy-related proteins (LC3, Atg-5, and Beclin-1) and augmented recruitment of phagosomal (EEA1 and Rab7) and lysosomal (LAMP1) proteins, thereby resulting in the reduction of the bacterial count in macrophages. The inhibition of phago-lysosome fusion by LprEwas found to be due to downregulation of IL-12 and IL-22 cytokines. Altogether, our data indicate that LprEis an important virulence factor that plays a crucial role in mycobacterial pathogenesis in the context of innate immunity.