Us animal experiments, it may be that a legumain DNA vaccine could be used to treat cancer in the humans. However, there are several issues preventing the clinical application of this potentially powerfultherapeutic strategy. One significant obstacle is 22948146 the lack of a suitable carrier [14]. Oral vaccination has advantages over intravenous administration, as it is noninvasive, more convenient, and achieves better clinical compliance [15]. In terms of biological carriers, both viral vectors and bacteria are used for oral DNA vaccination [16,17]. Bacteria-based delivery systems have been shown to be more effective in priming immune responses, able to be loaded with larger amounts of DNA clones, and more easily controlled compared with viral vectors [13]. Nonetheless, safety remains an issue when applying them to the humans. Hence, studies have been carried out to develop novel delivery carriers with low toxicity and high efficiency. In the past decade, chitosan nanoparticles (C.NPs) have emerged as novel carrier candidate because of their excellent stability, capacity to purchase Nafarelin enhance mucosa absorption, and good compatibility with vaccine DNA [18,19]. As a natural biopolymer derived from crustacean shells, C.NPs possess ideal properties of polymericChitosan NPs Loaded with 
Legumain DNA Vaccinecarriers; they are biocompatible, biodegradable, non-toxic, and inexpensive [20,21]. Studies show that C.NPs less than 500 nm in diameter are transported through the intestinal mucosa via an endocytotic mechanism [18,19,20]. Moreover, improved mucoadhesion and transient opening of tight junctions in the mucosal cell membrane contribute to the absorption-promoting effect of chitosan. By incorporating DNA plasmids into C.NP systems, oral DNA vaccines can be delivered to major targets, the buy Fruquintinib Peyer’s patches, and be taken up by antigen-presenting cells [18,20,22,23,24]. Although C.NPs have numerous advantages as delivery carriers for oral DNA vaccination, DNA degradation in the gut and low uptake efficiency in the gastrointestinal lymphoid tissue largely hamper their development. Recently, a delivery system composed of a chitosan core coated with sodium alginate has been described [25]. Borges et al. demonstrated that sodium alginate-coated nanoparticles were readily taken up by rat Peyer’s patches. Release profiles showed that burst release of loaded ovalbumin in pH of 1.2 (simulated gastric fluid) was largely prevented by its entrapment in alginate-coated chitosan nanoparticles. 15755315 This delivery system also acted as an effective adjuvant for hepatitis B surface antigen when subcutaneously administered in a mouse model [26]. Here, we hypothesized that alginic acid-coated chitosan nanoparticles (A.C.NPs) loaded with DNA plasmid could resist DNA degradation in the acidic gastric environment and be effectively taken up and expressed by antigen-presenting cells in the Peyer’s patches. In addition, we examined the therapeutic effect of a legumain DNA vaccine using this delivery system in a mouse breast cancer model.a mixture of oxygen/isoflurane before each experiment to alleviate their suffering. For survival data, humane endpoints were chosen to terminate the distress of the experimental animal via carbone dioxide euthanasia. The animals were monitored twice per day and extreme anorexia (poor appetite and emaciated appearance) was determined as a humane endpoint.Polymers and ReagentsChitosan (C-3646, MW: 810,000; degree of deacetylation 75?85 ) [27] and alginic acid (A.Us animal experiments, it may 
be that a legumain DNA vaccine could be used to treat cancer in the humans. However, there are several issues preventing the clinical application of this potentially powerfultherapeutic strategy. One significant obstacle is 22948146 the lack of a suitable carrier [14]. Oral vaccination has advantages over intravenous administration, as it is noninvasive, more convenient, and achieves better clinical compliance [15]. In terms of biological carriers, both viral vectors and bacteria are used for oral DNA vaccination [16,17]. Bacteria-based delivery systems have been shown to be more effective in priming immune responses, able to be loaded with larger amounts of DNA clones, and more easily controlled compared with viral vectors [13]. Nonetheless, safety remains an issue when applying them to the humans. Hence, studies have been carried out to develop novel delivery carriers with low toxicity and high efficiency. In the past decade, chitosan nanoparticles (C.NPs) have emerged as novel carrier candidate because of their excellent stability, capacity to enhance mucosa absorption, and good compatibility with vaccine DNA [18,19]. As a natural biopolymer derived from crustacean shells, C.NPs possess ideal properties of polymericChitosan NPs Loaded with Legumain DNA Vaccinecarriers; they are biocompatible, biodegradable, non-toxic, and inexpensive [20,21]. Studies show that C.NPs less than 500 nm in diameter are transported through the intestinal mucosa via an endocytotic mechanism [18,19,20]. Moreover, improved mucoadhesion and transient opening of tight junctions in the mucosal cell membrane contribute to the absorption-promoting effect of chitosan. By incorporating DNA plasmids into C.NP systems, oral DNA vaccines can be delivered to major targets, the Peyer’s patches, and be taken up by antigen-presenting cells [18,20,22,23,24]. Although C.NPs have numerous advantages as delivery carriers for oral DNA vaccination, DNA degradation in the gut and low uptake efficiency in the gastrointestinal lymphoid tissue largely hamper their development. Recently, a delivery system composed of a chitosan core coated with sodium alginate has been described [25]. Borges et al. demonstrated that sodium alginate-coated nanoparticles were readily taken up by rat Peyer’s patches. Release profiles showed that burst release of loaded ovalbumin in pH of 1.2 (simulated gastric fluid) was largely prevented by its entrapment in alginate-coated chitosan nanoparticles. 15755315 This delivery system also acted as an effective adjuvant for hepatitis B surface antigen when subcutaneously administered in a mouse model [26]. Here, we hypothesized that alginic acid-coated chitosan nanoparticles (A.C.NPs) loaded with DNA plasmid could resist DNA degradation in the acidic gastric environment and be effectively taken up and expressed by antigen-presenting cells in the Peyer’s patches. In addition, we examined the therapeutic effect of a legumain DNA vaccine using this delivery system in a mouse breast cancer model.a mixture of oxygen/isoflurane before each experiment to alleviate their suffering. For survival data, humane endpoints were chosen to terminate the distress of the experimental animal via carbone dioxide euthanasia. The animals were monitored twice per day and extreme anorexia (poor appetite and emaciated appearance) was determined as a humane endpoint.Polymers and ReagentsChitosan (C-3646, MW: 810,000; degree of deacetylation 75?85 ) [27] and alginic acid (A.