University of Groningen
Paving the way for pulmonary influenza vaccines
Tomar, Jasmine
IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below.
Document Version
Publisher's PDF, also known as Version of record
Publication date: 2018
Link to publication in University of Groningen/UMCG research database
Citation for published version (APA):
Tomar, J. (2018). Paving the way for pulmonary influenza vaccines: Exploring formulations, models and site of deposition. University of Groningen.
Copyright
Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).
Take-down policy
If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim.
Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons the number of authors shown on this cover page is limited to 10 maximum.
Introduction and outline of this thesis
Chapter 1
10
1
11
1
Introduction
Influenza, a highly contagious disease has caused high morbidity and mortality around the globe[1–4]. Vaccination is the best strategy to prevent the outbreaks of
seasonal influenza epidemics and sporadic pandemics in the human population. Currently, marketed influenza vaccine formulations are administered via injection, with an exception to Flumist® which is administered via the intranasal route.
Though immunization via injection is considered to be the gold standard, there are various drawbacks associated with injectable formulations. Challenges such as the requirement of trained healthcare personnel, needle phobia, pain and redness at the site of injection and transmission of infectious diseases during needle stick injuries make injectable preparations a bane rather than a boon[5–8]. Besides these
challenges, vaccination via injection does not induce an immune response at the portal of influenza virus entry, the respiratory tract[2,9–11]. Administration of the
vaccine via the respiratory tract can evade all the issues associated with delivery via injection as it is needle free, can be done by individuals themselves and can potentially elicit local immune responses[12,13].
Respiratory tract delivery, in particular pulmonary delivery, of liquid influenza vaccine formulations has already been successfully explored in the 1960’s[14,15]. For long this
research has not been continued, possibly because no convenient devices for pulmonary administration of liquid formulations were available at that time. Over the years, however, several of such devices have been developed, but these were not considered to be suitable for mass vaccination[7]. For the dispersion of dry powder formulations,
better delivery devices such as the disposable dry powder inhalers (DPI) are available now. These DPI’s (Twincer, Torus) are low-cost, single use devices that might be suitable for mass vaccination in cases of an influenza epidemic or pandemic[7].
Respiratory tract immunization may sound to be a feasible alternative to injectable vaccines, however, certain complexities related to it need to be resolved to facilitate a step further towards the clinic.
Objective of this thesis
In this thesis, we have investigated novel aspects for enhancing immunogenicity of influenza vaccine candidates delivered via the respiratory tract.
Chapter 1
12
1
The two major topics that were investigated were:
a) which site of antigen deposition within the respiratory tract of different pre-clinical models (mouse, cotton rat) results in optimal immune responses.
b) whether adjuvantation of influenza vaccine formulations can boost the immune responses in diverse animal models (mouse, chicken) leading to protective immunity.
Outline of this thesis
In Chapter 2, we focused on the need for the development of dry influenza vaccines with an emphasis on the drying processes used for their production. Further, we discussed the immunogenicity evoked by dry influenza vaccine formulations upon administration via alternative routes such as skin or mucosal areas (intranasal, pulmonary, sublingual and buccal) in comparison to the traditional parenteral route of administration. Lastly, the challenges and future developments with respect to dry influenza vaccine formulations are discussed.
These dry powder influenza vaccine formulations were further investigated and compared with liquid influenza formulations in terms of their deposition site, immunogenicity and protective efficacy. In Chapter 3, we investigated the site of deposition of pulmonary delivered liquid and powder influenza vaccine formulations in cotton rats using a fluorescent label and an imaging system. Also, the influence of deposition site of these formulations on their immunogenicity was evaluated by comparing the immune responses induced by liquid and powder formulations with each other. Finally, the effect of deposition site of these formulations on their protective efficacy, was determined by comparing the lung virus titers and monitoring the clinical symptoms induced after lethal live virus challenge.
The findings of Chapter 3 were further studied in Chapter 4 by targeting dry powder vaccine formulations to different regions of the respiratory tract of mice. Influenza and hepatitis B vaccines were used as model vaccine candidates for diseases that do or do not spread via the respiratory tract, respectively. Powder formulations of influenza and hepatitis-B were targeted to trachea/central airways by using the Penn-Century insufflator whereas deep lungs were targeted using an in-house aerosol generator.
13
1
A fluorescent label and an imaging system were used to confirm the deposition pattern. The effect of trachea/central airways vs deep lung targeting on immunogenicity was studied for both vaccine candidates.
In Chapter 5 and Chapter 6 another strategy that potentially leads to enhanced
immunogenicity of influenza vaccines delivered via the respiratory tract is described. Precisely, in Chapter 5, we have explored the potential of Advax™, a particulate insoluble polymorph of inulin, as a mucosal adjuvant. Advax-adjuvanted influenza formulations were either used as such or formulated into dry powder formulation for administration via the respiratory tract. The pharmaceutical aspects of a dry formulation were explored. Along with it, the immunological responses induced in mice upon administration via the respiratory tract (intranasal and pulmonary) were also investigated. In addition, we also investigated mechanistic insights related to respiratory tract administration of Advax-adjuvanted influenza vaccine. Finally, single-dose pulmonary immunized animals were challenged with a lethal single-dose of live virus to investigate the role of Advax-adjuvantation in protection.
In Chapter 6 the development of dry powder influenza vaccine formulations that can be inhaled by chickens is described. In order to protect millions of poultry animals against bird flu, an appropriate approach could be to aerosolize a dry powder formulation in a field where it can be inhaled by animals. Such situation was mimicked by aerosolizing dry powder influenza vaccine formulations in a box, in which chickens were able to inhale the aerosolized vaccine during breathing. The vaccine formulations were non-adjuvanted or adjuvanted with Advax or bacterium like particles. After immunization, the animals were challenged with a lethal dose of highly pathogenic avian influenza virus and were monitored for survival and shedding of challenge virus in their choanal and cloacal swabs. In addition, the immune responses induced by adjuvantation of influenza vaccine with bacterium like particles or Advax were also investigated.
Finally, in Chapter 7, the findings of this thesis are discussed and perspectives on respiratory tract delivery of vaccine formulations are presented.
Chapter 1
14
1
References
[1] WHO Report. Influenza (seasonal) fact sheet. World Health Organization, Geneva, Switzerland. 2016. [2] Bhide Y, Tomar J, Dong W, et al. Pulmonary delivery of influenza vaccine formulations in cotton rats:
site of deposition plays a minor role in the protective efficacy against clinical isolate of H1N1pdm virus. Drug Deliv. 2018; 25: 533–45.
[3] Kondrich J, Rosenthal M. Influenza in children. Curr Opin Pediatr. 2017; 29: 297–302. [4] Ghebrehewet S, MacPherson P, Ho A. Influenza. BMJ. 2016; 355: i6258.
[5] Cook IF. Evidence based route of administration of vaccines. Hum. Vaccin. 2008; 4: 67–73. [6] Amorij JP, Hinrichs WLJ, Frijlink HW, et al. Needle-free influenza vaccination. Lancet Infect. Dis.
2010; 10: 699–711.
[7] Tonnis WF, Lexmond AJ, Frijlink HW, et al. Devices and formulations for pulmonary vaccination. Expert Opin. Drug Deliv. 2013; 10: 1383–97.
[8] Gill HS, Kang S-M, Quan F-S, et al. Cutaneous immunization: an evolving paradigm in influenza vaccines. Expert Opin. Drug Deliv. 2014; 11: 615–27.
[9] Amorij J-P, Saluja V, Petersen A. H, et al. Pulmonary delivery of an inulin-stabilized influenza subunit vaccine prepared by spray-freeze drying induces systemic, mucosal humoral as well as cell-mediated immune responses in BALB/c mice. Vaccine. 2007; 25: 8707–17.
[10] Audouy SAL, van der Schaaf G, Hinrichs WLJ, et al. Development of a dried influenza whole inactivated virus vaccine for pulmonary immunization. Vaccine. 2011; 29: 4345–52.
[11] Patil H, Herrera Rodriguez J, de Vries-Idema J, et al. Adjuvantation of Pulmonary-Administered Influenza Vaccine with GPI-0100 Primarily Stimulates Antibody Production and Memory B Cell Proliferation. Vaccines. 2017; 5: 19.
[12] Belyakov IM, Ahlers JD. What Role Does the Route of Immunization Play in the Generation of Protective Immunity against Mucosal Pathogens? J. Immunol. 2009; 183: 6883–92.
[13] Tonnis WF, Kersten GF, Frijlink HW, et al. Pulmonary Vaccine Delivery: A Realistic Approach? J. Aerosol Med. Pulm. Drug Deliv. 2012; 25: 249–60.
[14] Waldman RH, Mann J, Small PA. Immunization Against Influenza, Prevention of Illness in Man by Aerosolized Inactivated Vaccine. 1969; 207: 520–24.
[15] Waldman RH, Bond JO, Levitt LP, et al. An evaluation of influenza immunization: influence of route of administration and vaccine strain. Bull. World Health Organ. 1969; 41: 543.
