Since antigens are negatively charged, they combine well with positively charged adjuvants. Here, ovalbumin (OVA) (0.1 mg·mL −1 ) and poly (diallyldimethylammonium chloride) (PDDA) (0.01 mg·mL −1 ) yielded PDDA/OVA assemblies characterized by dynamic light scattering (DLS) and scanning electron microscopy (SEM) as spherical nanoparticles (NPs) of 170 ± 4 nm hydrodynamic diameter, 30 ± 2 mV of zeta-potential and 0.11 ± 0.01 of polydispersity. Mice immunization with the NPs elicited high OVA-specific IgG1 and low OVA-specific IgG2a production, indicating a Th-2 response. Delayed-type hypersensitivity reaction (DTH) was low and comparable to the one elicited by Al(OH) 3 /OVA, suggesting again a Th-2 response. PDDA advantages as an adjuvant were simplicity (a single-component adjuvant), low concentration needed (0.01 mg·mL −1 PDDA) combined with antigen yielding neglectable cytotoxicity, and high stability of PDDA/OVA dispersions. The NPs elicited much higher OVA-specific antibodies production than Al(OH) 3 /OVA. In vivo, the nano-metric size possibly assured antigen presentation by antigen-presenting cells (APC) at the lymph nodes, in contrast to the location of Al(OH) 3 /OVA microparticles at the site of injection for longer periods with stimulation of local dendritic cells. In the future, it will be interesting to evaluate combinations of the antigen with NPs carrying both PDDA and elicitors of the Th-1 response.Vaccines 2020, 8, 105 2 of 16 or cationic polymers on superparamagnetic iron oxide NPs [18] have also been proposed as effective micro or nanomaterials able to effectively interact with antigens and antigen-presenting cells (APC).Major mechanisms for taking up the cationic assemblies depend on size [19,20]. Nanoparticles with 20-200 nm mean diameter are like viruses and usually taken up by endocytosis via clathrin-coated vesicles, caveolae, or their independent receptors, and preferentially ingested by dendritic cells (DC) [21]. Microparticles with 500-5000 nm diameter are like bacteria, captured by phagocytosis and primarily ingested by macrophages. All particles used in vaccine formulations are consequently internalized efficiently by APC by one or a combination of the quoted mechanisms [22,23]. Particles with diameters smaller than 500 nm, in particular the nano-sized ones with 40-100 nm diameter, are more efficient to promote CD8 and CD4 type 1 T-helper cells responses than those with diameters above 500 nm; large particles, more similar to Al (OH) 3 , usually induce good T-helper cells type 2 improvement in antibody responses [22]. In summary, cationic micro and nanoparticles are effectively taken up both by macrophages and dendritic cells since electrostatic attraction promotes particle binding and subsequent internalization [11,14,20,24,25]. Cationic particles and liposomes containing dioctadecyldimethylammonium bromide (DODAB) cationic lipid [13,26] carrying Mycobacterium tuberculosis [13,27], Chlamydia trachomatis [11], or Neisseria meningitides antigens enhanced the cellular and humoral immune re...