ARTICLE IN PRESS
G Model JVAC 14777 1–7
Vaccine xxx (2013) xxx–xxx
Contents lists available at ScienceDirect
Vaccine journal homepage: www.elsevier.com/locate/vaccine
Phase I/II trial of a replication-deficient trivalent influenza virus vaccine lacking NS1夽
1
2
3
Q1
4
a
5 6
Christina Mössler a,∗ , Franz Groiss a , Michael Wolzt b , Markus Wolschek a , Joachim Seipelt a , Thomas Muster a
Q2
b
AVIR Green Hills Biotechnology, 1200 Vienna, Austria Medical University of Vienna, Department of Clinical Pharmacology, 1090 Vienna, Austria
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8
a r t i c l e
i n f o
a b s t r a c t
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Article history: Received 12 August 2013 Received in revised form 13 October 2013 Accepted 14 October 2013 Available online xxx
14
21
Keywords: Influenza NS1 Intranasal Trivalent Live-attenuated Reverse genetics
22
1. Introduction
15 16 17 18 19 20
23 24 25 26 27 28 29 30 31 32 33 34
Background: The non-structural protein NS1 of the influenza virus counteracts the interferon-mediated immune response of the host. We investigated the safety and immunogenicity of a trivalent formulation containing influenza H1N1, H3N2 and B strains lacking NS1 (delNS1-trivalent). Methods: Healthy adult study participants who were seronegative for at least one strain present in the vaccine formulation were randomized to receive a single intranasal dose of delNS1-trivalent vaccine at 7.0 log10 TCID50/subject (n = 39) or placebo (n = 41). Results: Intranasal vaccination with the live replication-deficient delNS1-trivalent vaccine was well tolerated with no treatment-related serious adverse events. The most common adverse events identified, i.e. headache, oropharyngeal pain and rhinitis-like symptoms, were mainly mild and transient and distributed similarly in the treatment and placebo groups. Significant vaccine-specific immune responses were induced. Pre-existing low antibody titers or seronegativity for the corresponding vaccine strain yielded better response rates. Conclusions: We show that vaccination with a replication-deficient trivalent influenza vaccine containing H1N1, H3N2 and B strains lacking NS1 is safe and induces significant levels of antibodies (ClinicalTrials.gov identifier NCT01369862). © 2013 Published by Elsevier Ltd.
Influenza virus infection is a highly contagious and severe viral infection. Every year, influenza viruses infect 10–20% of the world population, causing up to 500,000 deaths in people of all ages [1]. Groups particularly at risk are older people, persons with chronic diseases, pregnant women and children [2,3]. In view of these data and the threat posed by potentially pandemic avian influenza strains, the European Influenza Surveillance Scheme (EISS) and/or the National Vaccine Program Office (NVPO) in the United States (US) seek to control the disease by using existing and developing new vaccines. Traditional trivalent inactivated vaccines (TIV) either produced in embryonated hen’s eggs or in certified cell lines are by far the most widely used vaccines for annual
夽 The design and results of the described clinical study were presented at the World Influenza Congress 2011 in Vienna, December 6th–8th 2011. ∗ Corresponding author. Current address: Global Research and Development, Baxter BioScience, WagramerStraße 17-19, 1220 Vienna, Austria. Tel.: +43 1 20100 2472549. E-mail address: christina [email protected] (C. Mössler).
immunization to prevent seasonal influenza infection [4]. However, there are ongoing efforts to develop novel types of vaccines including live attenuated influenza vaccines (LAIV) administered intranasally, which have already been used in Russia since 1988 [5] and in the US since 2003 [6]. Although serum antibody levels achieved by live vaccines are lower than those reached by inactivated vaccines, cold-adapted vaccines were shown to be comparable to conventional inactivated influenza vaccines in terms of preventing viral infection in adults [7,8] and proved to be even more effective in very young children aged 6–59 months [9]. However, the cold-adapted vaccine has a number of limitations; the most crucial is its ability to shed from vaccinated to non-vaccinated individuals [10]. We developed a novel type of live attenuated influenza vaccine by deleting the interferon antagonist NS1 (“delNS1”) from each of the influenza strains. DelNS1 mutants are capable of infecting respiratory epithelium cells after nasal administration but undergo abortive replication without forming progeny infectious particles. A first-in-man study with a monovalent delNS1-H1N1 vaccine candidate confirmed the tolerability and immunogenic potential of delNS1 vaccines [11]. Subsequently, we performed a first clinical study with a trivalent delNS1 vaccine candidate containing
0264-410X/$ – see front matter © 2013 Published by Elsevier Ltd. http://dx.doi.org/10.1016/j.vaccine.2013.10.061
Please cite this article in press as: Mössler C, et al. Phase I/II trial of a replication-deficient trivalent influenza virus vaccine lacking NS1. Vaccine (2013), http://dx.doi.org/10.1016/j.vaccine.2013.10.061
35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56
G Model JVAC 14777 1–7
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ARTICLE IN PRESS C. Mössler et al. / Vaccine xxx (2013) xxx–xxx
2
the H1N1, H3N2 and B strains recommended by the WHO for 2008/2009 to determine the safety and immunogenicity of a trivalent formulation in adults.
60
2. Material and methods
61
2.1. Study design
73
We did a randomized, double-blind, placebo-controlled phase I/II study in healthy males and females aged 18–60. The study was conducted at a single center located at the Medical University of Vienna, Austria, between 18 January 2011 and 25 August 2011. On study day 0, all participants received one dose of a trivalent delNS1 vaccine candidate containing the H1N1, H3N2 and B strains recommended by the WHO for 2008/2009 (delNS1-trivalent) or placebo at a 1:1 ratio. The study was conducted in compliance with Good Clinical Practice guidelines and the Declaration of Helsinki. It was approved by the independent ethics committee and the Austrian regulatory authority.
74
2.2. Vaccine and placebo
62 63 64 65 66 67 68 69 70 71 72
75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91
92
93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113
DelNS1-trivalent was generated by reverse genetics as described elsewhere [12–15]. Each dose contained 7.0 log10 fluorescent tissue culture infectious doses (fTCID50) of each of the three vaccine strain reassortants lacking the complete NS1 open reading frame and containing the hemagglutinin and neuraminidase of influenza A/Brisbane/59/07(H1N1)-like virus, A/Brisbane/10/07(H3N2)-like virus and B/Florida/04/06-like virus. The vaccine was produced in Vero cells under serumfree conditions following good manufacturing practice (GMP). Depending on the vaccine strain, the virus harvest was purified in one or two chromatographic steps and formulated with sucrose/phosphate/glutamate and recombinant serum albumin as stabilizer. The formulation buffer was given as placebo. Both vaccine and placebo were stored frozen at ≤−60 ◦ C and transferred into a nasal spray device (Pfeiffer, Germany) before intranasal administration. A total of 520 l of vaccine or placebo was administered into both nostrils of each participant (260 l/nostril). 2.3. Study population Healthy males and females aged 18–60, who were seronegative for one or two of the applied vaccine strains (i.e. antibody titers
G Model JVAC 14777 1–7
Vaccine xxx (2013) xxx–xxx
Contents lists available at ScienceDirect
Vaccine journal homepage: www.elsevier.com/locate/vaccine
Phase I/II trial of a replication-deficient trivalent influenza virus vaccine lacking NS1夽
1
2
3
Q1
4
a
5 6
Christina Mössler a,∗ , Franz Groiss a , Michael Wolzt b , Markus Wolschek a , Joachim Seipelt a , Thomas Muster a
Q2
b
AVIR Green Hills Biotechnology, 1200 Vienna, Austria Medical University of Vienna, Department of Clinical Pharmacology, 1090 Vienna, Austria
7
8
a r t i c l e
i n f o
a b s t r a c t
9 10 11 12 13
Article history: Received 12 August 2013 Received in revised form 13 October 2013 Accepted 14 October 2013 Available online xxx
14
21
Keywords: Influenza NS1 Intranasal Trivalent Live-attenuated Reverse genetics
22
1. Introduction
15 16 17 18 19 20
23 24 25 26 27 28 29 30 31 32 33 34
Background: The non-structural protein NS1 of the influenza virus counteracts the interferon-mediated immune response of the host. We investigated the safety and immunogenicity of a trivalent formulation containing influenza H1N1, H3N2 and B strains lacking NS1 (delNS1-trivalent). Methods: Healthy adult study participants who were seronegative for at least one strain present in the vaccine formulation were randomized to receive a single intranasal dose of delNS1-trivalent vaccine at 7.0 log10 TCID50/subject (n = 39) or placebo (n = 41). Results: Intranasal vaccination with the live replication-deficient delNS1-trivalent vaccine was well tolerated with no treatment-related serious adverse events. The most common adverse events identified, i.e. headache, oropharyngeal pain and rhinitis-like symptoms, were mainly mild and transient and distributed similarly in the treatment and placebo groups. Significant vaccine-specific immune responses were induced. Pre-existing low antibody titers or seronegativity for the corresponding vaccine strain yielded better response rates. Conclusions: We show that vaccination with a replication-deficient trivalent influenza vaccine containing H1N1, H3N2 and B strains lacking NS1 is safe and induces significant levels of antibodies (ClinicalTrials.gov identifier NCT01369862). © 2013 Published by Elsevier Ltd.
Influenza virus infection is a highly contagious and severe viral infection. Every year, influenza viruses infect 10–20% of the world population, causing up to 500,000 deaths in people of all ages [1]. Groups particularly at risk are older people, persons with chronic diseases, pregnant women and children [2,3]. In view of these data and the threat posed by potentially pandemic avian influenza strains, the European Influenza Surveillance Scheme (EISS) and/or the National Vaccine Program Office (NVPO) in the United States (US) seek to control the disease by using existing and developing new vaccines. Traditional trivalent inactivated vaccines (TIV) either produced in embryonated hen’s eggs or in certified cell lines are by far the most widely used vaccines for annual
夽 The design and results of the described clinical study were presented at the World Influenza Congress 2011 in Vienna, December 6th–8th 2011. ∗ Corresponding author. Current address: Global Research and Development, Baxter BioScience, WagramerStraße 17-19, 1220 Vienna, Austria. Tel.: +43 1 20100 2472549. E-mail address: christina [email protected] (C. Mössler).
immunization to prevent seasonal influenza infection [4]. However, there are ongoing efforts to develop novel types of vaccines including live attenuated influenza vaccines (LAIV) administered intranasally, which have already been used in Russia since 1988 [5] and in the US since 2003 [6]. Although serum antibody levels achieved by live vaccines are lower than those reached by inactivated vaccines, cold-adapted vaccines were shown to be comparable to conventional inactivated influenza vaccines in terms of preventing viral infection in adults [7,8] and proved to be even more effective in very young children aged 6–59 months [9]. However, the cold-adapted vaccine has a number of limitations; the most crucial is its ability to shed from vaccinated to non-vaccinated individuals [10]. We developed a novel type of live attenuated influenza vaccine by deleting the interferon antagonist NS1 (“delNS1”) from each of the influenza strains. DelNS1 mutants are capable of infecting respiratory epithelium cells after nasal administration but undergo abortive replication without forming progeny infectious particles. A first-in-man study with a monovalent delNS1-H1N1 vaccine candidate confirmed the tolerability and immunogenic potential of delNS1 vaccines [11]. Subsequently, we performed a first clinical study with a trivalent delNS1 vaccine candidate containing
0264-410X/$ – see front matter © 2013 Published by Elsevier Ltd. http://dx.doi.org/10.1016/j.vaccine.2013.10.061
Please cite this article in press as: Mössler C, et al. Phase I/II trial of a replication-deficient trivalent influenza virus vaccine lacking NS1. Vaccine (2013), http://dx.doi.org/10.1016/j.vaccine.2013.10.061
35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56
G Model JVAC 14777 1–7
57 58 59
ARTICLE IN PRESS C. Mössler et al. / Vaccine xxx (2013) xxx–xxx
2
the H1N1, H3N2 and B strains recommended by the WHO for 2008/2009 to determine the safety and immunogenicity of a trivalent formulation in adults.
60
2. Material and methods
61
2.1. Study design
73
We did a randomized, double-blind, placebo-controlled phase I/II study in healthy males and females aged 18–60. The study was conducted at a single center located at the Medical University of Vienna, Austria, between 18 January 2011 and 25 August 2011. On study day 0, all participants received one dose of a trivalent delNS1 vaccine candidate containing the H1N1, H3N2 and B strains recommended by the WHO for 2008/2009 (delNS1-trivalent) or placebo at a 1:1 ratio. The study was conducted in compliance with Good Clinical Practice guidelines and the Declaration of Helsinki. It was approved by the independent ethics committee and the Austrian regulatory authority.
74
2.2. Vaccine and placebo
62 63 64 65 66 67 68 69 70 71 72
75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91
92
93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113
DelNS1-trivalent was generated by reverse genetics as described elsewhere [12–15]. Each dose contained 7.0 log10 fluorescent tissue culture infectious doses (fTCID50) of each of the three vaccine strain reassortants lacking the complete NS1 open reading frame and containing the hemagglutinin and neuraminidase of influenza A/Brisbane/59/07(H1N1)-like virus, A/Brisbane/10/07(H3N2)-like virus and B/Florida/04/06-like virus. The vaccine was produced in Vero cells under serumfree conditions following good manufacturing practice (GMP). Depending on the vaccine strain, the virus harvest was purified in one or two chromatographic steps and formulated with sucrose/phosphate/glutamate and recombinant serum albumin as stabilizer. The formulation buffer was given as placebo. Both vaccine and placebo were stored frozen at ≤−60 ◦ C and transferred into a nasal spray device (Pfeiffer, Germany) before intranasal administration. A total of 520 l of vaccine or placebo was administered into both nostrils of each participant (260 l/nostril). 2.3. Study population Healthy males and females aged 18–60, who were seronegative for one or two of the applied vaccine strains (i.e. antibody titers
