The chemokine CCL4 (Chemokine C-C motif ligand 4), previously named Macrophage Inflammatory Protein-1β (MIP-1β), has an important role in inflammation and chemotaxis of different leukocyte populations during acute bacterial infections. In the present work, the authors evaluated the use of purified recombinant chicken CCL4, cloned and expressed in E. coli, as a chemotaxis agent to attract monocytes and neutrophils in vitro. Monocytes and neutrophils (also known as heterophils in chickens) were isolated from chicken blood, after separation with Histopaque 1107 and 1189 (Sigma, USA), respectively. The Boyden chamber (Millicell, Millipore, USA) was used to evaluate migration of monocytes and heterophils in the presence of rCCL4 in RPMI culture media. The role of rCCL4 in vivo was also evaluated. rCCL4 was mixed to sodium alginate in the presence of calcium chloride (1M) to encapsulate in microspheres, which were orally administered to 21 day old chickens (n=10) during 3 consecutive days (10mg/chicken). On the fourth day, all chickens were challenged with 108 CFU of Salmonella Typhimurium. At 2, 4 and 6 days post-infection (dpi), the control of the challenge bacteria was evaluated by plating and counting the CFU/g (Log10) and the population of macrophages and CD8+ T cells were evaluated by immunohistochemistry, in comparison with a negative group (untreated) in caecal tonsil sections. In vitro assay, showed a strong activity of rCCL4 on the attraction of monocytes and heterophils (p<0.05), with confirmed migration of these cells through the chamber membrane. In vivo assay, showed a significant and faster reduction of Salmonella Typhimuirium shedding in the feces and a significantly higher population of macrophages and CD8+ T cells (p<0.05) in the caecal tonsils of treated chickens at 4 and 6 dpi, when the bacterial numbers reached 2.4 and 2.0 CFU/g in treated animals against 3.8 and 3.5 CFU/g in untreated group, respectively.