Protection against Lethal Influenza with a Viral Mimic

Baker, Steven F.; Guo, Hailong; Albrecht, Randy A.; Garcı́a-Sastre, Adolfo; Topham, David J.; Martínez-Sobrido, Luis

doi:10.1128/jvi.01081-13

Cited by 60 publications

(104 citation statements)

References 37 publications

(54 reference statements)

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“…All mouse experiments were approved by the University Committee of Animal Resources and complied with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Research Council. Infections were performed as previously described (19). Briefly, mice were anesthetized intraperitoneally (i.p.)…”

Section: Methodsmentioning

confidence: 99%

“…The pCAGGS X31 HA and pCB7 hygromycin B resistance plasmids were used to cotransfect MDCK cells using Lipofectamine 2000 transfection reagent (Invitrogen) at a ratio of 3:1, as previously described (19). Cell clones were selected after serial dilution and testing for complementation of sciIV infection and immunofluorescence assay (IFA).…”

Section: Methodsmentioning

confidence: 99%

“…One advantage of the single-cycle replicating virus is that it can trigger effective CD8 T cell responses (14,16,17). To fully minimize the potential damage to the host, while maintaining the ability to stimulate CD8 T cell immunity, we generated a singlecycle infectious influenza X31 virus (X31-sciIV) using plasmidbased reverse-genetics techniques that we have described previously (18)(19)(20)(21). This virus has been modified so that the genomic HA segment, the receptor binding and fusion protein, has been replaced with green fluorescent protein (GFP).…”

mentioning

confidence: 99%

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Induction of CD8 T Cell Heterologous Protection by a Single Dose of Single-Cycle Infectious Influenza Virus

Guo

Baker

Martínez-Sobrido

et al. 2014

J Virol

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The effector functions of specific CD8 T cells are crucial in mediating influenza heterologous protection. However, new approaches for influenza vaccines that can trigger effective CD8 T cell responses have not been extensively explored. We report here the generation of single-cycle infectious influenza virus that lacks a functional hemagglutinin (HA) gene on an X31 genetic background and demonstrate its potential for triggering protective CD8 T cell immunity against heterologous influenza virus challenge. In vitro, X31-sciIV can infect MDCK cells, but infectious virions are not produced unless HA is transcomplemented. In vivo, intranasal immunization with X31-sciIV does not cause any clinical symptoms in mice but generates influenza-specific CD8 T cells in lymphoid (mediastinal lymph nodes and spleen) and nonlymphoid tissues, including lung and bronchoalveolar lavage fluid, as measured by H2-Db NP366 and PA224 tetramer staining. In addition, a significant proportion of X31-sciIV-induced antigen-specific respiratory CD8 T cells expressed VLA-1, a marker that is associated with heterologous influenza protection. Further, these influenza-specific CD8 T cells produce antiviral cytokines when stimulated with NP366 and PA224 peptides, indicating that CD8 T cells triggered by X31-sciIV are functional. When challenged with a lethal dose of heterologous PR8 virus, X31-sciIV-primed mice were fully protected from death. However, when CD8 T cells were depleted after priming or before priming, mice could not effectively control virus replication or survive the lethal challenge, indicating that X31-sciIV-induced memory CD8 T cells mediate the heterologous protection. Thus, our results demonstrate the potential for sciIV as a CD8 T cell-inducing vaccine. IMPORTANCEOne of the challenges for influenza prevention is the existence of multiple influenza virus subtypes and variants and the fact that new strains can emerge yearly. Numerous studies have indicated that the effector functions of specific CD8 T cells are crucial in mediating influenza heterologous protection. However, influenza vaccines that can trigger effective CD8 T cell responses for heterologous protection have not been developed. We report here the generation of an X31 (H3N2) virus-derived single-cycle infectious influenza virus, X31-sciIV. A one-dose immunization with X31-sciIV is capable of inducing functional influenza virusspecific CD8 T cells that can be recruited into respiratory tissues and provide protection against lethal heterologous challenge. Without these cells, protection against lethal challenge was essentially lost. Our data indicate that an influenza vaccine that primarily relies on CD8 T cells for protection could be developed.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Induction of CD8 T Cell Heterologous Protection by a Single Dose of Single-Cycle Infectious Influenza Virus

Guo

Baker

Martínez-Sobrido

et al. 2014

J Virol

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“…b Eight hemagglutinating units (HAU) of the indicated virus was incubated with 2-fold serial dilutions of the indicated sera. Plaque phenotypes were determined by immunostaining using the NP-specific monoclonal antibody HT103 as described previously (12,30). (C) Multicycle growth curve experiments were performed with MDCK cells as described previously (30).…”

mentioning

confidence: 99%

Development of a Mouse-Adapted Live Attenuated Influenza Virus That PermitsIn VivoAnalysis of Enhancements to the Safety of Live Attenuated Influenza Virus Vaccine

Cox

Baker

Nogales

et al. 2015

J Virol

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The live attenuated influenza virus vaccine (LAIV) is preferentially recommended for use in persons 2 through 49 years of age but has not been approved for children under 2 or asthmatics due to safety concerns. Therefore, increasing safety is desirable. Here we describe a murine LAIV with reduced pathogenicity that retains lethality at high doses and further demonstrate that we can enhance safety in vivo through mutations within NS1. This model may permit preliminary safety analysis of improved LAIVs. Influenza A virus is a respiratory pathogen that infects through the upper airway and leads to pathology via replication in the lower airway (1). The temperature gradient between these two areas in people enabled the development of the cold-adapted, live attenuated influenza virus vaccine (LAIV [FluMist]) that replicates in the cooler upper respiratory tract to trigger a protective immune response but cannot damage the lower respiratory tract due to the elevated temperatures restricting replication (2). This temperature-sensitive (ts) attenuated (att) phenotype is imparted by five mutations within the viral replicative machinery: namely, PB2 N265S, PB1 K391E, D581G, and A661T, and NP D34G (3, 4). Although this vaccine has an overall acceptable safety profile, it is not approved for use in children under 2 years of age due to concerns about elevated hospitalizations due to wheezing (5, 6). For this reason, it is also not approved for use in asthmatics. Therefore, development of vaccines with increased safety over LAIV is desirable. Currently, no mouse model exists for the adequate assessment of the safety of the LAIV. In experimental animal studies with LAIV, elevated doses of LAIV do not elicit pathology, rendering determination of safety impossible (7-10). Here, we describe a model with which we can assess alterations in vaccine safety.The parental strain for our vaccine is a well-characterized, murine-lethal strain of influenza A virus (A/Puerto Rico/8/34 H1N1, PR8). We introduced four ts att mutations from LAIV into PR8 (NP D34G is natively present) via site-directed mutagenesis (Agilent) and rescued this virus using plasmid-based reverse genetics techniques (11). This ts att virus (referred to henceforth as "PR8 LAIV") has been previously characterized in cell culture, but its phenotype in mice was not demonstrated (8). PR8 wild-type (WT) virus has a 50% lethal dose (LD 50 ) in C57BL/6 (B6) mice of 10 to 25 PFU (12, 13). Thus, we sought to ascertain the LD 50 of PR8 LAIV (Fig. 1). Groups of mice (n ϭ 5) were intranasally

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“…The capacity of antiviral prophylaxis to block or mitigate virus transmissibility has been studied by treating inoculated (68) or contact (32) ferrets with oseltamivir (Tamiflu). Vaccination or prior infection of donor ferrets can be useful when determining the efficacy of virus transmission under immune pressure (studied with both homologous and heterologous challenge viruses) (81)(82)(83)(84)(85) or when determining the efficacy of novel vaccine or antiviral candidates to limit transmission (68,86). These and other modifications to transmission schemes illustrate the high plasticity of this model and utility of these data for numerous applications beyond initial virus characterization but, given the wide range of alterations possible, similarly underscore the need for precision in how these data are presented and contextualized.…”

Section: Transmission Experimental Designsmentioning

confidence: 99%

Complexities in Ferret Influenza Virus Pathogenesis and Transmission Models

Belser

Eckert

Tumpey

et al. 2016

Microbiol Mol Biol Rev

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SUMMARYFerrets are widely employed to study the pathogenicity, transmissibility, and tropism of influenza viruses. However, inherent variations in inoculation methods, sampling schemes, and experimental designs are often overlooked when contextualizing or aggregating data between laboratories, leading to potential confusion or misinterpretation of results. Here, we provide a comprehensive overview of parameters to consider when planning an experiment using ferrets, collecting data from the experiment, and placing results in context with previously performed studies. This review offers information that is of particular importance for researchers in the field who rely on ferret data but do not perform the experiments themselves. Furthermore, this review highlights the breadth of experimental designs and techniques currently available to study influenza viruses in this model, underscoring the wide heterogeneity of protocols currently used for ferret studies while demonstrating the wealth of information which can benefit risk assessments of emerging influenza viruses.

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Protection against Lethal Influenza with a Viral Mimic

Cited by 60 publications

References 37 publications

Induction of CD8 T Cell Heterologous Protection by a Single Dose of Single-Cycle Infectious Influenza Virus

Induction of CD8 T Cell Heterologous Protection by a Single Dose of Single-Cycle Infectious Influenza Virus

Development of a Mouse-Adapted Live Attenuated Influenza Virus That PermitsIn VivoAnalysis of Enhancements to the Safety of Live Attenuated Influenza Virus Vaccine

Complexities in Ferret Influenza Virus Pathogenesis and Transmission Models

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