In previous researches, the results showed that Alhagi honey polysaccharide-loaded poly(D,L-lactic-co-glycolic) acid nanoparticles (AHPP) as immune adjuvant enhanced Th1 immune responses. In order to further enhance the immune adjuvant activity and phagocytosis of the nanoparticles, three kinds of Alhagi honey polysaccharide-loaded cationic polymer modified PLGA nanoparticles were prepared to investigate the effects on macrophages in vitro. After treatment with the nanoparticles, the effects of phagocytosis, co-stimulatory molecules expression, nitric oxide (NO), inducible nitric oxide synthase (iNOS), and cytokines secretion were evaluated. The results showed that the surface structure of cationic polymer modified AHPP nanoparticles were not obviously changed, and the stability was greatly improved. Cationic polymer modified AHPP nanoparticles significantly stimulated phagocytic activity, MHCII(+), CD86(+), and CD80(+) expression of macrophages. In addition, the levels of NO, iNOS, TNF-α, IFN-γ, IL-1β and IL-12 were enhanced in the peritoneal macrophages by stimulation with cationic polymer modified AHPP nanoparticles. Among them, polyethyleneimine modified PLGA nanoparticles (PEI-AHPP) showed the best effects on the expression of co-stimulatory molecules, and secretions of NO, iNOS, and cytokines. These results indicated that PEI-AHPP could enhance the activation of macrophages, and it could be potentially used as an AHP delivery system for the induction of strong immune responses.