Tissue resident macrophages (MTRs) regulate tissue repair and homeostasis by clearing cell debris, and are thought to form the first line of defense against pathogens. MTRs form early in life and self-renew locally. However, during tissue damage or disease, bone-marrow derived monocytes enter tissue sites and differentiate into MTRs, repairing the tissue and replenishing macrophages in the niche. How MTR versus monocyte-derived macrophages recruited during inflammation contribute to health and disease, and the cell-intrinsic mechanisms that control the monocyte to MTR transition across tissues, remain elusive. Here we show that deoxyhypusine synthase (DHPS), an enzyme that mediates the spermidine-dependent hypusine modification of the translation factor eIF5A, is required for the differentiation and maintenance of MTR. EIF5A is the only protein to contain hypusine, and the only function of DHPS (together with DOHH) is to hypusinate eIF5A. Hypusinated-eIF5A enhances the translation efficiency of certain mRNA transcripts that lead to ribosome stalling, including those with polyproline motifs. MTRs in tissues of young mice with myelomonocytic cell deletions in DHPS (Dhps-M mice; hypusination deficient eIF5A) had abnormal expression of macrophage tissue resident markers, and MTRs themselves declined with age. Single cell transcriptional analysis of DHPS-deficient peritoneal macrophages indicated a block in the transition from monocyte to mature MTRs, while proteomics revealed decreased expression of cell adhesion and signaling molecules. Notably, sequencing of ribosome-engaged transcripts suggested that certain cell adhesion and signaling molecules are hyper-dependent on hypusinated eIF5A for efficient translation.