. 2020 Nov 13;8:558349. doi: 10.3389/fbioe.2020.558349

TABLE 1.

Chitosan and modified chitosan nanoparticle-based vaccines: physicochemical properties, immune responses, and efficacy induced by vaccines administered to mucosal sites to protect against bacterial and viral infectious pathogens of poultry.

Vaccine carrier	Physicochemical properties of chitosan NPs based vaccine	Target pathogens	Encapsulated vaccine antigens	Route of delivery; antigen amount; number of vaccine doses	Target animal	Immune correlates and pathogen clearance	References
Chitosan NPs surface tagged with Salmonella Enteritidis flagellin protein	Low molecular weight chitosan Size: 517 nm Zeta potential: — Shape: spherical	Salmonella Enteritidis	Outer membrane proteins and flagellin protein	Oral; 100 μg; 3	Layer chickens	Enhanced mucosal IgA antibody, cellular immune response, TLRs gene expression	Renu et al., 2020c
Chitosan NPs surface tagged with Salmonella Enteritidis flagellin protein	Low molecular weight chitosan; 75–85% deacetylated; 50,000–190,000 Da Size: 514 nm Zeta potential: +40 mV Shape: spherical	Salmonella Enteritidis	Outer membrane proteins and flagellin protein	Oral; 50, 100 and 500 μg; 3	Layer chickens	Increased TLRs, Th1 and Th2 cytokines mRNA expression, antigen-specific humoral immune response; lower bacterial challenge load in cecum	Renu et al., 2020b
Chitosan NPs surface tagged with Salmonella Enteritidis flagellin protein	Low molecular weight chitosan Size: — Zeta potential: — Shape: —	Salmonella Enteritidis	Outer membrane proteins and flagellin protein	Oral; 500, 1000 and 2000 μg; 2	Broiler chickens	Induced cross-reactive IgG and mucosal IgA antibodies, cytokine gene expression; lower heterologous challenge bacterial load in liver and spleen	Acevedo-Villanueva et al., 2020
Chitosan NPs surface tagged with Salmonella Enteritidis flagellin protein	Low molecular weight chitosan Size: 300 nm Zeta potential: +40 mV Shape: spherical	Salmonella Enteritidis	Outer membrane proteins and flagellin protein	Oral; 12.5 and 50 μg; 1 and 2	White Cornish Cross broilers	Increased innate immune response and antigen specific lymphocytes proliferation; lower antigen specific IgA and IgG antibody response	Han et al., 2020c
Chitosan NPs surface tagged with Salmonella Enteritidis flagellin protein	Low molecular weight chitosan Size: — Zeta potential: — Shape: —	Salmonella Enteritidis	Outer membrane proteins and flagellin protein	Oral; 10 and 50 μg; 1 to 3	Cornish Cross breed broilers	Robust pre and post-challenge IgG and IgA antibody, cell mediated immune response, TLRs gene expression; reduced challenge bacterial load in the cecum	Han et al., 2020a
Mannose conjugated chitosan NPs surface tagged with Salmonella Enteritidis flagellin protein	Low molecular weight chitosan Size: — Zeta potential: — Shape: —	Salmonella Enteritidis	Outer membrane proteins and flagellin protein	Oral; 10 μg; 2	Cornish Cross breed broilers	Enhanced cell mediated immune response, TLRs and balanced Th1 and Th2 cytokine gene expression; reduced Salmonella challenge load in the cecum	Han et al., 2020b
Chitosan NPs	Size: 167 nm Zeta potential: +20.1 mV Shape: spherical	Campylobacter jejuni	Recombinant hemolysin co-regulated protein	Oral; 50 μg; 3	Vencobb chicks	Increased secretory IgA and systemic IgY antibody, cytokine gene expression; enhanced bacterial clearance in cecum	Singh et al., 2019
Chitosan NPs	85% deacetylated; 400 kD Size: 80–100 nm Zeta potential: — Shape: spherical	Campylobacter jejuni	flaA gene	Intranasal; 150 μg; 3	White Leghorn chickens	Increased serum IgG and intestinal mucosal IgA antibody; reduced bacterial shedding by 2–3 log₁₀ in large intestine and cecum	Huang et al., 2010
Chitosan NPs	Medium molecular weight chitosan; 75–85% deacetylated Size: — Zeta potential: +19.9 mV Shape: spherical	Avian pathogenic Escherichia coli	ΦKAZ14 bacteriophage	Oral; 10⁷ PFU/mL; 1	Broiler chicks	Improved body weight; decreased mortality; decreased bacterial colonization in intestines; reduced fecal shedding; increased protection rate	Kaikabo et al., 2017
Chitosan NPs surface tagged with HA2 and M2e influenza proteins	Low molecular weight chitosan Size: 100–800 nm Zeta potential: — Shape: spherical	Avian influenza virus	HA2 and M2e mRNAs	Intranasal; 4 μg; 2	Layer chickens	Increased IgG and mucosal IgA antibody, virus neutralization titers, cell-mediated immune response; lower lung pathology; reduced homologous and heterologous challenge virus titers in cloacal swab	Hajam et al., 2020
Chitosan	85% deacetylated Size: — Zeta potential: — Shape: —	Avian influenza virus	Inactivated split influenza virus	Intranasal; 100 HA units; 1 and 2	Layer chickens	Enhanced mucosal IgA and HI antibody; resist against lethal virus challenge in field condition	Worrall et al., 2009
Chitosan NPs	Medium molecular weight chitosan; 75–85% deacetylated Size: 286 nm Zeta potential: +19.9 mV Shape: spherical	Avian infectious bronchitis virus	Inactivated infectious bronchitis virus	Oculo-nasal; 10⁸.²⁸⁵ EID₅₀ of the virus; 1	SPF chickens	Enhanced mucosal IgA, IFNγ gene expression; no or mild relevant microscopic lesions; lower viral load in trachea and kidney	Lopes et al., 2018
N-2- hydroxypropyl trimethyl ammonium chloride chitosan and N,O-carboxymethyl chitosan NPs	Size: 251.8 and 122.4 nm Zeta potential: 46.6 and 53.2 mV Shape: spherical	Newcastle disease virus and infectious bronchitis virus	Newcastle disease virus and infectious bronchitis virus combined	Intranasal; 10⁷.⁴ and 10⁵.⁵ EID₅₀ of the virus; 1	SPF chickens	Increased IgG and IgA antibodytiters, lymphocyte proliferation, cytokines IL-2, IL-4 and IFN-γ than the commercial live vaccine; complete protection	Zhao et al., 2017
N-2-hydroxypropyl trimethyl ammonium chloride chitosan and N,O carboxymethyl chitosan NPs	Size: 309.7 nm Zeta potential: 49.9 mV Shape: spherical	Newcastle disease virus	pVAX I-F(o)	Intranasal; 200 μg; 2	SPF chickens	Higher IgG and secretory IgA antibody; stimulated lymphocyte proliferation; increased IL-2, IL-4, and IFN-γ levels; complete protection	Zhao et al., 2018
N-2-hydroxypropyl trimethyl ammonium chloride chitosan and N,O carboxymethyl chitosan NPs	85% deacetylated Size: 252.2 nm Zeta potential: +41.1 mV Shape: spherical	Newcastle disease virus	Attenuated live Newcastle disease virus	Oral or intranasal; 10⁷.⁵ EID₅₀ of the virus; 1	SPF chickens	Induced high titers of serum antibody; promoted lymphocyte proliferation; higher levels of serum IL-2, IL-4 and IFN- γ; no clinical signs and mortality	Jin et al., 2017
N-2-hydroxypropyl trimethyl ammonium chloride chitosan NPs	85% deacetylated; 71.3 kDa Size: 303.8 nm Zeta potential: 45.7 mV Shape: spherical	Newcastle disease virus	Attenuated live Newcastle disease virus	Oral or intranasal; 10⁷.¹² EID₅₀ of the virus; 1	SPF chickens	Induced robust cellular, humoral and mucosal immune response; absence of pathological changes; complete protection than the commercial attenuated live vaccine	Zhao et al., 2016
O-2’ hydroxypropyl trimethyl ammonium chloride chitosan NPs	85% deacetylated Size: 303.5 nm Zeta potential: +46.3 mV Shape: spherical	Newcastle disease virus	Attenuated live Newcastle disease virus	Oral or intranasal; —; 1	SPF chickens	Robust cellular, humoral and mucosal immune response; absence of histopathological changes compared to a commercial attenuated live vaccine	Dai et al., 2015
Chitosan NPs	80% deacetylated; 71.3 kDa Size: 199.5 nm Zeta potential: +12.1 mV Shape: spherical	Newcastle disease virus	F gene plasmid DNA	Intranasal; 200 μg; 2	SPF chickens	Increased IgA and IgG antibodies, lymphocyte proliferation response; absence of clinical symptoms and mortality	Zhao et al., 2014
Chitosan NPs	80% deacetylated; 71.3 kDa Size: 371.1 nm Zeta potential: +2.8 mV Shape: spherical	Newcastle disease virus	Lentogenic live Newcastle disease virus	Oral or intranasal; —; 1	SPF chickens	Enhanced lymphocyte proliferation, serum HI and intestinal mucus IgA antibody titers; absence of clinical signs and mortality	Zhao et al., 2012
Chitosan	70–95% deacetylated Size: — Zeta potential: — Shape: —	Newcastle disease virus	Live Newcastle disease virus	Oculo-nasal; 10⁶ EID₅₀ of the virus; 1	SPF white Leghorn chickens	Enhanced cell mediated immune response; no effect on systemic and mucosal antibody mediated immune response	Rauw et al., 2010a
Chitosan	—	Newcastle disease virus	Live Newcastle disease virus	Oculo-nasal; 10⁶ EID₅₀ of the virus; 1	Isa Brown layer chickens	Provides protection against mortality and morbidity; reduced virus shedding; higher cellular and mucosal antibody mediated immune response	Rauw et al., 2010b

NPs, nanoparticles; PFU, plaque forming units; HA, hemagglutinin; EID₅₀, 50% egg infective dose; HI, hemagglutination inhibition; TLRs, toll-like receptors; Th1, T helper type 1; Th2, T helper type 2; IL, Interleukin; and IFNγ, interferon gamma.