Dry influenza vaccines: towards a stable, effective and convenient alternative to conventional parenteral influenza vaccination

Figures & data

Figure 1. Publications found on dry influenza vaccines per route of administration expressed as percentage. Numbers are based on literature found from 2000 to 2015 using Embase and Pubmed.

Table 1. An overview of stability and immunogenicity of dry influenza vaccines described in literature.

	Formulation
Delivery route	Antigen	Excipient(s)	Adjuvant	Drying process	Stability data	Delivery device	Species	Immune responses	Reference
Dermal	WIV	Trehalose	¯	AD		PowderJect^®	BALB/c mice	Significantly high HI titers; protected against viral challenge	[45]
	Split	Trehalose	+	AD		PowderJect^®	BALB/c mice	Significantly high HI titers; better protected against viral challenge	[47]
	NP peptide, WIV	PEG	¯	VD		PowderJect XR-1	BALB/c mice	NP: Strong antibody and CTL responses; complete protection against viral challenge	[48]
	Split	Trehalose	+	AD		PowderJect ^®	BALB/c mice	Strong serum, mucosal and HI titers; protection against lethal viral challenge	[49]
	WIV	Trehalose, Mannitol, Dextran	+	SFD	Preserved potency at 40°C/75% RH/4 months	PowderJect ^®	BALB/c mice; Rhesus Macaques	Significantly high HI titers; QS-21 augmented immune response in monkeys protects against lethal challenge	[46]
	WIV	Trehalose	+	AD		PowderJect ^®	BALB/c mice	Strong HI titers; cytokine production by epidermal cells treated with LTR72	[50]
	WIV	Trehalose, Mannitol, Dextran	¯	SFD		PowderJect ND 5.2	Healthy adults	Equivalent or high HI titers and seroconversion	[9]
	WIV	CMC, Lutrol, Trehalose	¯	AD		Coated MN	BALB/c mice	Equivalent humoral and cellular responses; complete protection against challenge	[51]
	WIV	CMC, Lutrol, Trehalose	¯	AD	Preserved 65% HA activity after coating	Coated MN	BALB/c mice	Stable MN formulations: higher humoral and recall immune responses	[52]
	WIV	CMC, Lutrol, Trehalose	¯	AD		Coated MN	BALB/c mice	Strong humoral, cellular and recall responses; rapid viral clearance	[53]
	WIV	CMC, Lutrol	¯	AD		Coated MN	BALB/c mice	Equivalent humoral responses and protective efficacy against viral challenge	[54]
	VLP	Trehalose	¯	AD	Preserved 58% HA activity after coating	Coated MN	BALB/c mice	Significantly high humoral recall responses and better protection	[55]
	VLP	CMC, Lutrol, Trehalose	¯	AD		Coated MN	BALB/c mice	Low dose MN as immunogenic as high dose i.m.. Better protective efficacy of MN coated formulations	[56]
	VLP	CMC, Lutrol, Trehalose	¯	AD		Coated MN	BALB/c mice	Enhanced humoral and recall responses; better protection against viral challenge	[57]
Dermal	WIV	PVP	¯	FD		Dissolvable MN	BALB/c mice	Better humoral, cellular responses, enhanced cellular recall responses; better protection against viral challenge	[58]
	VLP	CMC, AA, PVP, XG, Lutrol, Trehalose, Sucrose, Dextran, Inulin	¯	AD	Preserved more than 60% HA activity with CMC/Trehalose	Coated MN	BALB/c mice	Stable MN formulations: potent antibody responses, protection against lethal challenge compared to unstabilized formulations	[59]
	Split	MC, Trehalose	¯	GJD	Preserved stability at 23°C/6 months	Coated NP	C57BL/6 mice	Dose sparing effect: NP generated equivalent protective immune responses with 1/30th dose	[60]
	WIV	CMC, Lutrol, Trehalose	¯	AD		Coated MN	BALB/c mice	Long-lasting immunity and complete protection against viral challenge after 6 months	[61]
	Split	MC	+	GJD		Coated NP	C57BL/6 mice	Dose sparing effect: up to 900-fold dose sparing by co-delivery of Ag and adjuvant by NP	[62]
	Subunit	CMC, Lutrol, Trehalose	¯	AD		Coated MN	BALB/c mice	Long-lasting immunity and protective HI titers; drop in HI titer and partial protection in the i.m. group	[63]
	Split	MC	¯	GJD		Coated NP	C57BL/6 mice	Equivalent antigen migration and antibody generation kinetics	[64]
	WIV	CMC, Lutrol, Trehalose	¯	AD	Reduced HA activity with increase in crystallization and phase separation of coating	Coated MN	BALB/c mice	Decreased immunogenicity of crystallized and phase separated coatings compared to fresh stable coatings	[13]
	HA DNA vaccine	None	¯	AD		Coated MN	BALB/c mice	Potent humoral, cellular responses and long-lasting immunity	[65]
	HA DNA vaccine	CMC, Lutrol	¯	AD	Viscosity enhancer/surfactant affects stability of DNA vaccine	Coated MN	BALB/c mice	Strong humoral and better protective responses	[66]
	WIV	CMC, Lutrol, Trehalose	¯	AD	Preserved 20% HA activity at 25°C/1 month	Coated MN	BALB/c mice	Stable MN formulations: strong humoral responses and better protection than unstabilized MN	[67]
Dermal	WIV	CMC, Lutrol, Trehalose	¯	AD	Preserved antigen stability by surfactant and viscosity enhancer combination	Coated MN	BALB/c mice	Stabilizer/viscosity enhancer combination augments antibody response and complete viral protection compared to unstabilized formulations	[68]
	WIV, HA DNA	HA DNA, Trehalose	¯	AD	Preserved 100% HA activity in the presence of DNA as viscosity enhancer	Coated MN	BALB/c mice	Strong homologous and heterologous antibody responses; better protection against challenge	[69]
	Subunit	Sucrose	¯	FD	Stable at 4°C and RT/dessicant for 2 months; good freeze–thaw stability	Coated MN	Guinea pigs	Equivalent HI titers	[70]
	VLP	CMC, Lutrol, Trehalose	¯	AD		Coated MN	BALB/c mice	Stable MN formulations: Strong systemic, mucosal and recall responses 14 months after vaccination compared to unstabilized MN formulations	[71]
	WIV	CMC, Trehalose, Sucrose, fish gelatin	¯	AD	Preserved antigenic activity at RT/3 months	Dissolvable MN	BALB/c mice	Better humoral responses	[72]
	Subunit	CMC, Lutrol, Trehalose	¯	AD		Coated MN	BALB/c mice	Strong antibody titers, reduced viral titers; better viral protection	[73]
	Live, WIV	CMC, Trehalose, XG, SA	¯	AD	Preserved antigenic stability by sugar/viscosity enhancer	Coated MN	BALB/c mice	Stronger antibody titers elicited by stabilized formulations than unstabilized formulations	[74]
	M2e-flagellin	CMC, Lutrol	¯	AD		Coated MN	Mice	Strong antibody titers and better protection	[75]
	M2e-VLPs	CMC, Lutrol, Trehalose	¯	AD	Preserved antigenic stability/immunogenicity 8 weeks at room temperature	Coated MN	BALB/c mice	Strong antibody titers and better cross protected	[16]
	Subunit	Hyaluronate, Dextran, Povidone	¯	DD	Preserved antigenic stability for 6 months at 4°C	Dissolvable MN	Healthy adults	Equivalent or stronger immune responses against A and B strains, respectively	[17]
Nasal	WIV	Trehalose, Chitosan	¯	FD	Antigen retains stability at 25°C/25% RH for 12 weeks	In-house device	Brown Norway Rats	Higher nasal mucosal and equivalent systemic immune responses	[76]
	WIV	Trehalose, Chitosan, SA, CMC, HPMC	¯	SFD	Antigen retains stability at 25°C/40% RH for 12 weeks	In-house device	Brown Norway Rats	Comparable systemic and better mucosal responses	[77]
	Split	Lutrol, PVP, HPMC, Carbopol, Chitosan, XG, SDS, Carrageenan	+	FD	Antigen’s stability was reduced by xanthan gum and CL	Inserts	OF-1 mice and Sprague-Dawley Rats	XG inserts with CL-induced comparable immune responses to i.n. liquid formulations	[78]
	WIV	Chitosan, TPP, Tween 80, Span 80	+	VD		In-house device	Rabbits	Adjuvanted vaccine elicited significantly higher systemic and mucosal responses	[79]
Pulmonary	Subunit	Inulin	¯	SFD	Preserved antigenic stability after SFD	Penn-Insufflator	BALB/c mice	Significantly superior humoral and cell-mediated immune responses	[80]
	Subunit	Inulin	¯	SD/SFD	Antigen stable at 20°C/3 years	Penn-Insufflator	BALB/c mice	Significantly high IgG titers	[24]
	WIV	Inulin	¯	SFD	Preserved antigenic stability after SFD	Penn-Insufflator	BALB/c mice	Potent systemic and mucosal responses; equivalent viral reduction in lungs	[81]
	WIV	Inulin	¯	SFD	Preserved antigenic stability at 30°C/3 months	Penn-Insufflator	BALB/c mice	Strong systemic and mucosal responses; slightly decreased immunogenicity of stored formulations compared to those freshly prepared	[11]
	WIV	Inulin	¯	SFD	Preserved antigenic stability after SFD	Penn-Insufflator	Chickens	Strong HI titers; complete protection against lethal challenge	[82]
	WIV	Inulin	+	SFD	Preserved activity of WIV/MPLA after SFD	Penn-Insufflator	BALB/c mice	Potent systemic and mucosal responses of adjuvanted formulations; equivalent magnitude of viral protection	[83]
	WIV	Inulin	+	SFD	Preserved activity of WIV/adjuvants after SFD	Penn-Insufflator	BALB/c mice	Potent systemic and mucosal responses; partial protection against heterologous challenge	[84]
Oral/Sublingual/Buccal	Split	MC	¯	GJD		Coated NP	C57BL/6 mice	Equivalent/Better systemic immune responses to parenteral/oral formulations	[19]
	HA DNA Vaccine	Cellulose, Starch, Eudragit 100	+	FD		Self-administered tablets	Healthy adults	Significant increase in HI and MN titers as compared to placebo group	[20]

Immunogenicity is compared to parenteral influenza vaccines unless stated otherwise. Plus and minus sign indicates with or without adjuvant, respectively.

NPP: nucleoproteinpeptide; WIV: whole inactivated virus; VLP: virus like particle; HA: hemagglutinin; SD: spray drying; AD: air drying; VD: vacuum drying; DD: dessicator drying; FD: freeze drying; SFD: spray freeze drying; GJD: gas jet drying; PEG: polyethylene glycol; CMC: carboxymethyl cellulose; PVP: polyvinylpyrrolidone; CT: cholera toxin; CpG DNA: synthetic oligodeoxynucleotide containing CpG motifs; AA: alginic acid; XG: xanthan gum; MC: methylcellulose; SA: sodium alginate; HPMC: hydroxypropylmethylcellulose; SDS: sodium lauryl sulfate TPP: tripolyphosphate; RH: relative humidity; RT: room temperature; MN: microneedles: HI: hemagglutination inhibition titers; CTL: cytotoxic T lymphocyte; NP: nanopatch; Ag: antigen; CL: cationic lipid; i.n.: intranasal; i.m.: intramascular.

Figure 2. (a) Formulation excipients required for the production of dry, stable influenza vaccines per route of administration. (b) A general overview of advantages and disadvantages associated with non-parenteral routes used for influenza vaccine immunization in preclinical and clinical studies.

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