Elsevier

Vaccine

Volume 29, Issue 21, 12 May 2011, Pages 3773-3781
Vaccine

Inactivated and live, attenuated influenza vaccines protect mice against influenza:Streptococcus pyogenes super-infections

https://doi.org/10.1016/j.vaccine.2011.03.031Get rights and content

Abstract

Mortality associated with influenza virus super-infections is frequently due to secondary bacterial complications. To date, super-infections with Streptococcus pyogenes have been studied less extensively than those associated with Streptococcus pneumoniae. This is significant because a vaccine for S. pyogenes is not clinically available, leaving vaccination against influenza virus as our only means for preventing these super-infections. In this study, we directly compared immunity induced by two types of influenza vaccine, either inactivated influenza virus (IIV) or live, attenuated influenza virus (LAIV), for the ability to prevent super-infections. Our data demonstrate that both IIV and LAIV vaccines induce similar levels of serum antibodies, and that LAIV alone induces IgA expression at mucosal surfaces. Upon super-infection, both vaccines have the ability to limit the induction of pro-inflammatory cytokines within the lung, including IFN-γ which has been shown to contribute to mortality in previous models of super-infection. Limiting expression of these pro-inflammatory cytokines within the lungs subsequently limits recruitment of macrophages and neutrophils to pulmonary surfaces, and ultimately protects both IIV- and LAIV-vaccinated mice from mortality. Despite their overall survival, both IIV- and LAIV-vaccinated mice demonstrated levels of bacteria within the lung tissue that are similar to those seen in unvaccinated mice. Thus, influenza virus:bacteria super-infections can be limited by vaccine-induced immunity against influenza virus, but the ability to prevent morbidity is not complete.

Introduction

Influenza A virus infections that are complicated by secondary bacterial pneumonia make a significant contribution to deaths during both influenza virus epidemics [1] and pandemics [2]. During typical influenza seasons, Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, and Streptococcus pyogenes [3], [4], [5] are all known contributors to the “excess mortality” that results from influenza virus super-infections. In fact, in the 1918–1919 influenza pandemic, S. pneumoniae and S. pyogenes (Group A Streptococcus) were the most frequently observed bacterial species in the lungs of infected soldiers [6], and together they likely contributed to as many as 90% of deaths attributed to this pandemic [2]. More recently, findings from the H1N1 swine-origin influenza virus pandemic demonstrate that 29% of deaths were due to secondary bacterial pneumonia in an autopsy series, with 27% of these fatalities being associated with S. pyogenes super-infection [7]. Furthermore, S. pneumoniae and S. pyogenes were the most frequent species associated with increased parapneumonic empyema in a study conducted during the 2009 H1N1 pandemic in Utah [8]. The incidence of invasive diseases caused by S. pyogenes in England increased significantly (26%) in December 2010 and January 2011 in all age groups, due, in part, to widespread influenza infections. Interestingly, the greatest percentage of invasive disease episodes associated with laboratory confirmed influenza infection during this period were caused by S. pyogenes [9].

Several studies have assessed the efficacy and effectiveness of influenza vaccines to prevent influenza-like illness [10], [11], [12], [13], but less information is available regarding the ability of influenza vaccines to limit secondary bacterial complications [14], [15], [16], [17]. Since secondary bacterial infections are the primary cause of mortality associated with influenza virus, methods to limit these complications are currently being sought [18]. The purpose of this study was to directly compare the contributions of IIV and LAIV toward protection in a murine model of influenza virus:S. pyogenes super-infection. We report that both IIV and LAIV vaccines induced systemic (serum) antibody responses, with LAIV also eliciting local (mucosal) IgA antibodies. Subsequently, mice vaccinated against influenza virus demonstrated reduced inflammatory cytokines within bronchoalveolar lavage fluid (BALF), decreased recruitment of inflammatory cells to the lungs, and increased survival, compared to unvaccinated control mice. Despite limiting mortality associated with these super-infections, similar levels of viable bacteria were detected within the lungs of both vaccinated and unvaccinated mice, an outcome that was not observed after sub-lethal inoculation with S. pyogenes alone. Thus, immunity induced after vaccination against influenza virus (either IIV or LAIV) prevented super-infections within mice, albeit incompletely. Overall, protection against super-infection was similar for recipients of either IIV or LAIV.

Section snippets

Mice

Adult (6–8-week-old) female BALB/cJ mice were obtained from Harlan Laboratories (Indianapolis, IN) and housed in groups of four, with 24-h access to food and water. All animal experiments were performed following the guidelines established and approved by the Animal Care and Use committee at the University of South Dakota (Vermillion, SD).

Super-infection model

Viruses expressing the hemagglutinin (HA) and neuraminidase (NA) from A/Hong Kong/1/68-H3N2 were created as described previously [19], [20], and this

Influenza virus:S. pyogenes super-infections can be modeled using an H3N2 HA-expressing virus and MGAS315 bacteria

We first established a murine model of influenza virus:S. pyogenes super-infection using a previously characterized strain of HK68 virus [19] and MGAS315 bacteria [23] for super-infection. Mice were divided into groups that received either HK68 virus alone, MGAS315 bacteria alone, or HK68 virus + MGAS315 bacteria. A control group that received neither virus nor bacteria was also included (Fig. 1). The 3 groups inoculated with either HK68 virus alone, MGAS315 bacteria alone, or HK68 virus + MGAS315

Discussion

The mortality associated with influenza is often due to secondary bacterial complications, including those caused by S. pyogenes [2], [7], [8]. In this study, we established a model of influenza:S. pyogenes super-infection and compared the independent contributions of IIV and LAIV vaccines toward preventing morbidity and mortality associated with this super-infection. Our results demonstrate that despite differences in vaccine-induced immunity, both vaccines provided significant, albeit

Acknowledgments

The authors recognize the technical contributions of Kara Pitchford and Kevin Cwach. We also thank Melanee Clark and the Animal Resource Center at USD and acknowledge the contributions of the Flow Cytometry Core Facility at Sanford Research, in particular Satoshi Nagata and John Lee. Finally, we thank Jonathan A. McCullers for providing HK68 viruses for use in these studies and for critically reading the manuscript.

Conflict of interest: The authors have no conflicts of interest to disclose.

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    Portions of this manuscript were presented at the Options for the Control of Influenza VII (September, 2010) in Hong Kong SAR (Abstract Number P-569) and the International Conference on Gram-Positive Pathogens (October, 2010) in Omaha, NE (Poster Number 40).

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