A practical influenza neutralization assay to simultaneously quantify hemagglutinin and neuraminidase-inhibiting antibody responses

Hassantoufighi, Arash; Zhang, Henry; Sandbulte, Matthew R.; Gao, Jin; Manischewitz, Jody; King, Lisa R.; Golding, Hana; Straight, Timothy M.; Eichelberger, Maryna C.

doi:10.1016/j.vaccine.2009.10.066

Cited by 54 publications

(51 citation statements)

References 26 publications

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“…Some GMTs shown in Table 2 were previously reported [20], but are repeated here to emphasize that the response rate measured by NI assay was not significantly different between vaccine types.…”

Section: Comparison Of Antibody Responses Following Receipt Of Live Amentioning

confidence: 71%

“…Hemagglutination Inhibition (HI) assay: HI titers were measured as previously described [20]. Briefly, sera were treated with receptor destroying enzyme and then heat-inactivated.…”

Section: Antibody Assaysmentioning

confidence: 99%

“…An accelerated viral inhibition assay with NA as read-out (AVINA assay) was used to measure titers as previously described [20]. Briefly, MDCK cells were washed in serum-free medium (EMEM containing glutamine, penicillin and streptomycin) and 50 μl of a 8 x 10 5 /ml cell suspension placed in wells of flat-bottomed 96-well plates.…”

Section: Avina Neutralization Assaymentioning

confidence: 99%

“…Response rates were greatest when antibody titers were measured by an AVINA neutralization assay, an assay that has excellent sensitivity and includes detection of functional antibodies with specificity for both HA and NA [20]. Even with this assay, there were significantly fewer responders to each of the 3 virus strains in LAIV than TIV ( Table 2).…”

Section: Comparison Of Antibody Responses Following Receipt Of Live Amentioning

confidence: 99%

“…Many of these subjects did not have a 4-fold increase in HI titer, and consideration of NI titers did not increase the percent of responding individuals. The use of AVINA, a sensitive neutralization assay developed in our laboratory [20], identified a greater number of responders in each vaccine group (Tables 2,4), with 75% of TIV recipients and 38% of LAIV recipients, responding to all 3 vaccine components. CD4 + T cell responses were more robust after LAIV than TIV (Figure 3), and inclusion of individuals with H3N2-specific T cell proliferation ≥2 fold the response prior to vaccination, resulted in the identification of 100% of TIV recipients and 63% of LAIV recipients as responders.…”

Section: Evaluation Of Laiv Immunogenicity In Adultsmentioning

confidence: 99%

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Qualitative Differences in T cell responses to Live, Attenuated and Inactivated Influenza Vaccines

Eichelberger¹,

Rivers²,

Ream³

et al. 2012

J Clin Cell Immunol

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Annual epidemics of influenza cause considerable morbidity and mortality. Trivalent inactivated vaccine (TIV) and live, attenuated influenza vaccine (LAIV) are licensed in the United States, and both are effective in preventing disease in persons younger than 49. Serum hemagglutination inhibition (HI) titers correlate with TIV but not LAIV efficacy, suggesting that additional effector mechanisms are induced to the live, attenuated vaccine and play an important role in protection against disease. For this reason there is a need to identify surrogate markers of LAIV efficacy that are easily measured in robust assays. We have compared the immunogenicity of TIV and LAIV in a small clinical study (16 age-matched volunteers in each vaccine group) by measuring serologic responses using traditional HI and NA inhibition assays as well as a sensitive cell-based neutralization assay. In addition, we evaluated cellular responses by measuring the quantity and quality of antigen-specific CD4+ and CD8 + T cells following vaccination. The quality of the CD4 + T cell response was different for each vaccination group, with CD4 + T cell proliferation and increased secretion of IFN-γ characteristic of responses following immunization with LAIV, while antigen-specific T cells that secreted IL-5 were more frequently measured from TIV recipients. Our results suggest that sensitive, serologic assays with broad specificity, together with CD4 + T cell proliferation and IFN-γ secretion provide a more complete measure of the immunogenicity of LAIV in adults, and could be used to enhance the identification of vaccine responders.

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Section: Comparison Of Antibody Responses Following Receipt Of Live Amentioning

confidence: 71%

“…Hemagglutination Inhibition (HI) assay: HI titers were measured as previously described [20]. Briefly, sera were treated with receptor destroying enzyme and then heat-inactivated.…”

Section: Antibody Assaysmentioning

confidence: 99%

Section: Avina Neutralization Assaymentioning

confidence: 99%

Section: Comparison Of Antibody Responses Following Receipt Of Live Amentioning

confidence: 99%

Section: Evaluation Of Laiv Immunogenicity In Adultsmentioning

confidence: 99%

See 3 more Smart Citations

Qualitative Differences in T cell responses to Live, Attenuated and Inactivated Influenza Vaccines

Eichelberger¹,

Rivers²,

Ream³

et al. 2012

J Clin Cell Immunol

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Contribution of antibody production against neuraminidase to the protection afforded by influenza vaccines

Marcelin

Sandbulte

Webby

2012

Reviews in Medical Virology

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SUMMARY Vaccines are instrumental in controlling the burden of influenza virus infection in humans and animals. Antibodies raised against both major viral surface glycoproteins, hemagglutinin (HA) and neuraminidase (NA), can contribute to protective immunity. Vaccine‐induced HA antibodies have been characterized extensively, and they generally confer protection by blocking the attachment and fusion of a homologous virus onto host cells. Although not as well characterized, some functions of NA antibodies in influenza vaccine‐mediated immunity have been recognized for many years. In this review, we summarize the case for NA antibodies in influenza vaccine‐mediated immunity. In the absence of well‐matched HA antibodies, NA antibodies can provide varying degrees of protection against disease. NA proteins of seasonal influenza vaccines have been shown in some instances to elicit serum antibodies with cross‐reactivity to avian‐origin and swine‐origin influenza strains, in addition to HA drift variants. NA‐mediated immunity has been linked to (i) conserved NA epitopes amongst otherwise antigenically distinct strains, partly attributable to the segmented influenza viral genome; (ii) inhibition of NA enzymatic activity; and (iii) the NA content in vaccine formulations. There is a potential to enhance the effectiveness of existing and future influenza vaccines by focusing greater attention on the antigenic characteristics and potency of the NA protein. Copyright © 2012 John Wiley & Sons, Ltd.

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Influenza Neuraminidase as a Vaccine Antigen

Eichelberger

Wan

2014

Current Topics in Microbiology and Immunology

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Neuraminidase (NA) is the second most abundant influenza surface glycoprotein and contributes to virus replication in several ways, most notably by removing sialic acids from the host and viral glycoproteins, releasing newly formed virus particles from infected cells. Antibodies that block this enzyme activity restrict virus replication in vitro. This chapter describes foundational epidemiologic and human influenza challenge studies that provide evidence of an association between NA inhibiting antibodies and resistance to disease. Mouse challenge studies show that while NA immunity is infection-permissive, NA-specific antibodies attenuate infection and prevent severe disease. NA immunity is most effective against homologous viruses but there is substantial protection against viruses with a heterologous NA (different lineage within a NA subtype). Monoclonal antibodies specific for conserved antigenic domains of subtype N1 protect against seasonal and pandemic H1N1 as well as H5N1 virus challenge. Clinical studies demonstrate that licensed seasonal vaccines contain immunogenic amounts of NA, but the contribution of this immunity to vaccine efficacy is currently not known. New types of influenza vaccines could be designed to elicit NA immunity. Because NA induces heterologous immunity, it could be an important constituent of universal influenza vaccines that aim to protect against unexpected emerging viruses.

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A practical influenza neutralization assay to simultaneously quantify hemagglutinin and neuraminidase-inhibiting antibody responses

Cited by 54 publications

References 26 publications

Qualitative Differences in T cell responses to Live, Attenuated and Inactivated Influenza Vaccines

Qualitative Differences in T cell responses to Live, Attenuated and Inactivated Influenza Vaccines

Contribution of antibody production against neuraminidase to the protection afforded by influenza vaccines

Influenza Neuraminidase as a Vaccine Antigen

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