Quaternized chitosan nanoparticles loaded with the combined attenuated live vaccine against Newcastle disease and infectious bronchitis elicit immune response in chicken after intranasal administration

Abstract

Newcastle disease (ND) and infectious bronchitis (IB) are important diseases, which cause respiratory diseases in chickens, resulting in severely economic losses in the poultry industry. In this study, N-2-hydroxypropyl trimethyl ammonium chloride chitosan (N-2-HACC) and N,O-carboxymethyl chitosan (CMC) were synthesized as adjuvant and delivery carrier for vaccine antigens. N-2-HACC-CMC/NDV/IBV nanoparticles (NPs) (NDV/La Sota and IBV/H120 encapsulated in N-2-HACC-CMC NPs) and N-2-HACC-CMC/NDV-IBV NPs (the mixing of N-2-HACC-CMC/NDV NPs and N-2-HACC-CMC/IBV NPs in a ratio of 1:1) were prepared by the polyelectrolyte composite method, respectively. Both nanoparticles exhibited lower cytotoxicity and higher stability. Their bioactivities were maintained when they were stored at 37 °C for three weeks. Release assay in vitro showed that both NDV and IBV could be sustainably released from the nanoparticles after an initial burst release. In vivo immunization of chickens showed that N-2-HACC-CMC/NDV/IBV NPs or N-2-HACC-CMC/NDV-IBV NPs intranasally induced higher titers of IgG and IgA antibodies, significantly promoted proliferation of lymphocytes and induced higher levels of interleukine-2 (IL-2), IL-4 and interferon-γ (IFN-γ) than the commercially combined attenuated live vaccine did. This is the first study in the field of animal vaccines demonstrating that intranasal administration of chickens with antigens (NDV and IBV) encapsulated with chitosan derivative could induce humoral, cellular, and mucosal immune responses, which protected chickens from the infection of highly virulent NDV and IBV. This study indicated that N-2-HACC-CMC could be used as an efficient adjuvant and delivery carrier for further development of mucosal vaccines and drugs and could have an immense application potential in medicine.

Chitosan Derivatives and Their Application in Biomedicine

“…N-2-HACC and CMCS were synthesized as vaccine adjuvants and delivery systems in our group, where an animal experiment showed that the intranasal N-2-HACC/CMCS nanoparticles produced higher IgG (immunoglobulin G) and sIgA (secretory immunoglobulin A) antibody titers and higher levels of cytokines than the commercially available vaccine did [209]. Additionally, our team also synthesized chitosan-coated PLGA(poly(lactic-co-glycolic acid) nanoparticles, and NDV (Newcastle disease virus) energy-containing DNA that was encapsulated in the nanoparticles was prepared to evaluate the mucosal immune response; the results indicated that the nanoparticles induced stronger cellular, humoral, and mucosal immune responses than the plasmid DNA alone, showing that the chitosan-coated PLGA nanoparticles could be used as an efficient delivery system for mucosal immunization in the case of DNA vaccines [210].…”

Applications of polymer-based nanoparticles in vaccine field

“…Presently, a variety of natural and synthetic biodegradable polymer-based nanoparticles can be used to enhance immunogenicity for vaccine delivery. In addition, vaccine antigens encapsulated in the polymer-based nanoparticles administered through the mucosal pathway can protect antigens from degradation and ensure that the encapsulated antigen is released at the action site to stimulate efficient immune responses [14]. Polymer-based nanoparticles have also exhibited great value and potential in improving the immunogenicity of vaccines and the development of nano vaccines.…”

Engineered Nanodelivery Systems to Improve DNA Vaccine Technologies

“…Chitosan has also been reported to possess natural adjuvant effects, capable of inducing DC maturation through the stimulation of type I interferon (IFN) release which has the added benefit of inducing antigen-specific Th1 responses [32]. Chitosan-based NP delivery technology has been developed over the years and has resulted in numerous developments including a potential therapeutic anti-tumour human papillomavirus (HPV) DNA vaccine [33], treatments for influenza A [34,35], viral myocarditis [36], and a host of animal-borne diseases including Newcastle disease virus (NDV) [37][38][39][40], Nodavirus [41], and Trueperella pyogenes [42]. As mentioned previously, one method to improve the efficacy of DNA vaccines is through targeted delivery to APCs which can be achieved through the functionalization of NPs.…”