The largest populations of the human virome belong to bacteriophages. M13 filamentous bacteriophages can survive, circulate and being cleared within the body. The M13 phage has been developed as a new therapeutic application due to its ability. Being detected by innate immune cells, M13 phage triggers an immune response. Hereupon, macrophages are counted as the first immune cells stimulated by phage particles. The possible interference between immune responses with the phage-induced treatment process, necessitates determining M13 phage effect on immune responses. For this purpose, M13 phage with RGD were used as substrates for macrophage culture. Gelatin and uncoated wells were used as controls. 2 and 7 days post culture on the coated surfaces, peritoneal macrophages were analyzed in terms of morphological characteristics and metabolic profile and measurement of cytokines production. Macrophages on M13 phage containing surfaces secreted anti-inflammatory cytokines, increased efferocytosis activity and show metabolic profile of an M2 macrophage. The results demonstrate that M13 phage can modulate immune responses by polarization of macrophages toward anti-inflammatory phenotype over time. Moreover, the combination of RGD peptide motif with M13 phage was able to stimulate macrophage polarization more effectively which may introduce an Immune-modulating technology for therapy and biocontrol.