Pandemic H1N1 vaccine requires the use of an adjuvant to protect against challenge in naïve ferrets

doi:10.1016/j.vaccine.2010.12.125

Vaccine

Volume 29, Issue 11, 3 March 2011, Pages 2120-2126

https://doi.org/10.1016/j.vaccine.2010.12.125 Get rights and content

Abstract

In the context of an A/H1N1 influenza pandemic situation, this study demonstrates that heterologous vaccination with an AS03-adjuvanted 2008/2009 seasonal trivalent and pandemic H5N1 monovalent split vaccine conferred partial protection in influenza-naïve ferrets after challenge with the influenza pandemic H1N1 A/The Netherlands/602/09 virus. Further, unlike saline control and non-adjuvanted vaccine, it was shown that immunization of naïve ferrets with an AS03-adjuvanted pandemic H1N1 A/California/7/09 influenza split vaccine induced increased antibody response and enhanced protection against the challenge strain, including significant reduction in viral shedding in the upper respiratory tract and reduced lung pathology post-challenge. These results show the need for vaccination with the adjuvanted vaccine to fully protect against viral replication and influenza disease in unprimed ferrets.

Introduction

An important hallmark of the recently emerged 2009 A/H1N1 pandemic influenza virus is that it causes disease in young people more often than it does in older individuals. Indeed, in contrast to seasonal influenza, a substantial proportion of the cases of severe illness and death have occurred among young people [1] who are generally devoid of cross-reactive antibody responses against 2009 A/H1N1, while 34% of people over 60 years exhibited cross-reactive seroprotection [2]. These observations suggest the need for a potent vaccine for the younger population in the context of the current 2009 A/H1N1 influenza pandemic due to the lack of preexisting immunity against this virus.

Vaccination is the most effective intervention to reduce morbidity and mortality during an influenza pandemic provided that vaccines are rapidly available in sufficient quantities. Based on early experience with non-adjuvanted H5N1 vaccines in a population largely naïve to the H5N1 strain, two vaccinations with high haemagglutinin (HA) antigen doses (up to 90 μg HA) were shown to be required to elicit putatively protective antibody levels [3]. However, the necessity for such high dose of antigen would lead to vaccine shortage when facing a pandemic. Therefore, the use of adjuvants was advocated to increase immunogenicity, with the aim to allow antigen sparing but also promote cross-immunity against drifted strains. Antigen sparing and inter-clade cross-reactivity was demonstrated by using recombinant clade 1 H5N1 A/Vietnam/1194/2004 formulated with AS03_A, an oil-in-water-based Adjuvant System containing α-tocopherol that stimulates antibody production [4], [5]. AS03-induced increased immunogenicity was also observed in a clinical trial involving H1N1 strain [6].

It is not ethically possible to study the efficacy of a pandemic vaccine in the target human population prior to the onset of the pandemic. Animal studies are thus required. In this regard, ferrets (Mustela putorius furo) are considered to be the best mammalian model to evaluate the efficacy of influenza vaccines for humans [7], [8]. In ferrets, the pandemic H1N1 A/The Netherlands/602/09 influenza virus is more pathogenic than seasonal influenza viruses and infects not only the upper airways (nasal passages) but also the more distal parts of the airways (bronchi and alveoli) [9], [10]. The severity of the pneumonia that the pandemic H1N1 causes approaches that caused by highly pathogenic avian influenza H5N1 virus, but the lethal dose of the pandemic H1N1 is higher compared to H5N1 [8], [10].

Currently, data obtained with 2009 A/H1N1 influenza vaccines suggested the importance of using an adjuvant and two vaccinations to induce adequate humoral responses in unprimed children [11], [12]. To investigate this hypothesis, the present study was designed to evaluate in naïve ferrets the efficacy of vaccination with a 2008/2009 seasonal trivalent split vaccine, an H5N1 pandemic split vaccine or the H1N1 A/California/7/09 split vaccine with or without AS03, against a challenge with the novel H1N1 A/The Netherlands/602/09 influenza virus.

Section snippets

Immunization with the seasonal trivalent split vaccine or H5N1 pandemic split vaccine

Eight groups of six ferrets (Mustela putorius furo) received intramuscularly one (day 21) or two vaccinations (days 0 and 21) with the seasonal trivalent split influenza vaccine (H1N1 A/Brisbane/59/07, H3N2 A/Uruguay/716/07 and B/Brisbane/3/07) containing 15, 7.5 or 3.75 μg HA/strain adjuvanted with AS03_A (oil-in-water emulsion-based Adjuvant System containing 11.86 mg tocopherol) or AS03_B (oil-in-water emulsion-based Adjuvant System containing 5.93 mg tocopherol) in the hind leg under ketamin (4–8

H1N1 challenge in ferrets immunized with adjuvanted or non-adjuvanted seasonal trivalent split vaccine or H5N1 pandemic split vaccine

We investigated a 2009 A/H1N1 challenge study in naïve ferrets that received one vaccination (15 μg HA/strain, group 3) or two vaccinations (15 μg HA/strain for groups 2 and 4, 7.5 μg HA/strain for the group 5 and 3.75 μg HA/strain for the group 6) with the 2008/2009 seasonal trivalent split influenza vaccine (H1N1 A/Brisbane/59/07, H3N2 A/Uruguay/716/07 and B/Brisbane/3/07) adjuvanted with AS03_A (groups 2, 3 and 6) or AS03_B (groups 4 and 5). One group of ferrets received two vaccinations with the

Discussion

The present study was designed to evaluate in naïve ferrets the protective efficacy against challenge with the novel H1N1 A/The Netherlands/602/09 influenza virus induced by one or two vaccinations with different doses of a 2008/2009 seasonal trivalent split vaccine (H1N1 A/Brisbane/59/07, H3N2 A/Uruguay/716/07 and B/Brisbane/3/07), an H5N1 pandemic split vaccine and finally an H1N1 A/California/7/09 split vaccine with or without AS03, an oil-in-water emulsion based Adjuvant System containing

Acknowledgments

The authors are very grateful to C. Dubois, P. Festraets, V. Gobin, Ph. Lancelot, M.-C. Mortier, B. Bouzya, V. Francq, A. Van Beneden, R. Mascolo, M. Hamrouni (GSK Biologicals, Rixensart, Belgium) and W. van Aert, R. Boom, C. van Hagen, R. van Lavieren, and P. van Run (ViroClinics Biosciences BV and Erasmus Medical Center, Rotterdam, The Netherlands) for their very precious technical support. Finally, we would like to thank U. Krause (GSK Biologicals, Rixensart, Belgium) and P. Cadot (XpePharma

References (30)

J.-L. Bresson et al.
Safety and immunogenicity of an inactivated split-virion influenza A/Vietnam/1194/2004 (H5N1) vaccine: phase I randomised trial
Lancet
(2006)
I. Leroux-Roels et al.
Antigen sparing and cross-reactive immunity with an adjuvanted rH5N1 prototype pandemic influenza vaccine: a randomised controlled trial
Lancet
(2007)
F. Roman et al.
Immunogenicity and safety in adults of one dose of influenza A H1N1v 2009 vaccine formulated with and without AS03_A-adjuvant: preliminary report of an observer-blind, randomised trial
Vaccine
(2010)
E. Plennevaux et al.
Immune response after a single vaccination against 2009 influenza A H1N1 in USA: a preliminary report of two randomised controlled phase 2 trials
Lancet
(2010)
J.H.C.M. Kreijtz et al.
Infection of mice with a human influenza A/H3N2 virus induces protective immunity against lethal infection with influenza A/H5N1 virus
Vaccine
(2009)
P. Vandepapelière et al.
Potent enhancement of cellular and humoral immune responses against recombinant hepatitis B antigens using AS02A adjuvant in healthy adults
Vaccine
(2005)
B. Baras et al.
Longevity of the protective immune response induced after vaccination with one or two doses of AS03A-adjuvanted split H5N1 vaccine in ferrets
Vaccine
(2011)
I. Leroux-Roels et al.
Priming with AS03_A-adjuvanted H5N1 influenza vaccine improves the kinetics, magnitude and durability of the immune response after a heterologous booster vaccination: an open non-randomised extension of a double-blind randomised primary study
Vaccine
(2010)
C. Fraser et al.
Pandemic potential of a strain of influenza A (H1N1): early findings
Science
(2009)
K. Hancock et al.
Cross-reactive antibody responses to the 2009 pandemic H1N1 influenza virus
N Engl J Med
(2009)

I. Leroux-Roels et al.

Broad Clade 2 cross-reactive immunity induced by an adjuvanted clade 1 rH5N1 pandemic influenza vaccine

PLoS One

(2008)

A.W. Hampson

Ferrets and the challenges of H5N1 vaccine formulation

J Infect Dis

(2006)

B. Baras et al.

Cross-protection against lethal H5N1 challenge in ferrets with an adjuvanted pandemic influenza vaccine

PLoS One

(2008)

V.J. Munster et al.

Pathogenesis and transmission of swine-origin 2009 A(H1N1) influenza virus in ferrets

Science

(2009)

J.M.A. van den Brand et al.

Severity of pneumonia due to new H1N1 influenza virus in ferrets is intermediate between that due to seasonal H1N1 virus and highly pathogenic avian influenza H5N1 virus

J Infect Dis

(2010)

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Nasal influenza vaccines composed of inactivated pH1N1/09 split or whole virus antigen mixed with Endocine™ adjuvant induced high antibody titers in influenza naïve ferrets and protection against homologous challenge. Nasal Endocine™ formulated inactivated pH1N1/09 influenza vaccines induced high levels of serum HI and VN antibodies, which were in the same order of magnitude as antibody levels found in influenza naïve ferrets intranasally immunized with monovalent H1N1/California/2009 live attenuated influenza vaccines (LAIVs) [37–39] or in influenza naïve ferrets intramuscularly immunized with AS03 adjuvanted H1N1/California/2009 split vaccines [39]. The results in control ferrets parenterally immunized with non-adjuvanted seasonal TIV were similar to those seen in naïve controls (i.n. saline).
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Pandemic H1N1 vaccine requires the use of an adjuvant to protect against challenge in naïve ferrets

Abstract

Introduction

Section snippets

Immunization with the seasonal trivalent split vaccine or H5N1 pandemic split vaccine

H1N1 challenge in ferrets immunized with adjuvanted or non-adjuvanted seasonal trivalent split vaccine or H5N1 pandemic split vaccine

Discussion

Acknowledgments

Lancet

Lancet

Vaccine

Lancet

Vaccine

Vaccine

Vaccine

Vaccine

Pandemic potential of a strain of influenza A (H1N1): early findings

Science

Cross-reactive antibody responses to the 2009 pandemic H1N1 influenza virus

N Engl J Med

Broad Clade 2 cross-reactive immunity induced by an adjuvanted clade 1 rH5N1 pandemic influenza vaccine

PLoS One

Ferrets and the challenges of H5N1 vaccine formulation

J Infect Dis

Cross-protection against lethal H5N1 challenge in ferrets with an adjuvanted pandemic influenza vaccine

PLoS One

Pathogenesis and transmission of swine-origin 2009 A(H1N1) influenza virus in ferrets

Science

Severity of pneumonia due to new H1N1 influenza virus in ferrets is intermediate between that due to seasonal H1N1 virus and highly pathogenic avian influenza H5N1 virus

J Infect Dis