Immunostimulatory effect of N-2-hydroxypropyltrimethyl ammonium chloride chitosan-sulfate chitosan complex nanoparticles on dendritic cells.

In this study, we synthesized negatively charged chitosan sulfate and positively charged hydroxypropyltrimethyl ammonium chloride chitosan (HACC), and then prepared chitosan derivatives with positive and negative ions as nanoparticles (NPs) by ovalbumin encapsulation using the polyelectrolyte method. NPs with different substitution sites and molecular weights (MW) were prepared by varying conditions. We then determined the zeta potential average, diameter, encapsulation effect, and their immunostimulatory effects on dendritic cells (DCs). The results showed that chitosan-derivative NPs ranged in size from 153.33 to 320.90 nm; all NPs were positive, with charges ranging from 17.10 to 39.30 mV and the encapsulation rates of 65 %-75 %. Three NPs greatly promoted the expression and secretion of interleukin-6 (IL-6), tumor necrosis factor (TNF-α), and interleukin-1β (IL-1β) in DC cells: C2,3,6 chitosan sulfate-HACC (C2,3,6-HACC; 200 kDa), C3,6 chitosan sulfate-HACC (C3,6-HACC; 200 kDa) and C6 chitosan sulfate-HACC (C6-HACC; 50 kDa). We also found that 200-kDa C2,3,6-HACC and 50-kDa C6-HACC NPs greatly increased secretion of the major histocompatibility complex-II (MHC-II), CD40, CD80, and CD86, indicating that these NPs promote effective antigen presentation, further increasing immunity effects. Finally, we applied laser confocal photography and determined that NPs entered the cell to promote the regulation of cellular immune activity; this discovery lays a foundation for further research on their mechanism of their action. Therefore, C2,3,6-HACC and C6-HACC NPs have the potential as immunological adjuvants.