Pseudoparticle neutralization is a reliable assay to measure immunity and cross-reactivity to H5N1 influenza viruses

Alberini, Isabella; Tordello, Elena Del; Fasolo, Alba; Temperton, Nigel; Galli, Grazia; Gentile, Chiara; Montomoli, Emanuele; Hilbert, Anne Katrin; Banzhoff, Angelika; Giudice, Giuseppe Del; Donnelly, John; Rappuoli, Rino; Capecchi, Barbara

doi:10.1016/j.vaccine.2009.07.079

Cited by 61 publications

(73 citation statements)

References 20 publications

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“…As already described in previous studies, owing to the poor immunogenicity of the HA of A/H5N1 and the immunologic naivety of the population, H5N1-specific antibodies can be measured by HI and MN assays mainly after the second vaccine dose of MF59-H5N1 vaccine, and only rarely functional antibodies are measurable at baseline [4,21]. However, using PNA, which is more sensitive than HI and MN assays [20], we detected neutralizing antibodies already at baseline in plasma of 42% of H1N1-High subjects, even though at low titers. On the contrary, plasma-neutralizing antibodies were present at baseline only in 8% of H1N1-Low subjects.…”

Section: Preexisting H1n1-igg Mbcs May Favor a Functional Response Tomentioning

confidence: 96%

“…Plasma samples were tested by PNA as previously described starting from a dilution 1:100, for 5 1:3 dilutions in duplicates [20]. Neutralization titers were determined as the dilution required to obtain 80% reduction in relative light units compared with control wells with virus alone.…”

Section: Pseudoparticle Neutralization Assaymentioning

confidence: 99%

“…To gain insight on whether neutralizing epitopes were included among those targeted by H5/H1 cross-reactive MBCs, we looked for H5-neutralizing antibodies in plasma samples and supernatants from MBC cultures with a pseudoparticle neutralization assay (PNA) [20] using pseudoviral particles expressing the HA from the A/Vietnam/1194/04 H5N1 strain. This assay not only provides information about the impact of a large H1N1-MBC preexistent pool on the development of neutralizing antibodies, but also restricts the effect to cross-reactivity to HA molecule, since the pseudoparticles only express the H5 HA, but not the NA.…”

Section: Preexisting H1n1-igg Mbcs May Favor a Functional Response Tomentioning

confidence: 99%

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Impact of preexisting memory to seasonal A/H1N1 influenza virus on the immune response following vaccination against avian A/H5N1 virus

et al. 2013

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Cross-protection against divergent strains of influenza virus is an objective of various vaccination approaches. B cells cross-neutralizing several influenza A heterosubtypes have been isolated from cultured human memory B cells (MBCs) and plasmablasts early after influenza vaccination or infection.However, a systematic assessment of the frequency of MBCs and plasmablasts in the blood of healthy individuals is lacking. Here, we show that under resting conditions about 45% of human adults never vaccinated nor exposed to avian A/H5N1 influenza have detectable circulating MBCs cross-reacting with H5N1. This proportion rises to 63.3% among subjects with a large pool of MBCs specific for seasonal H1N1 (i.e. frequency ≥1% of total IgG MBCs). Moreover, subjects with high baseline frequencies of H1N1-specific MBCs had an expansion of H5N1-specific MBCs producing H5-neutralizing antibodies already after the first dose of an MF59-adjuvanted H5N1 vaccine. These results suggest that H1N1-specific MBCs contain a subset of cells cross-reacting to H5. We propose that a proportion of human adults have a pool of H5/H1 cross-reactive MBCs that contribute to the rapid rise of the antibody response to divergent influenza strains. This may have implications on vaccination strategies aimed at counteracting future influenza pandemics. Keywords:Cross-reactive antibodies r H5N1 r Influenza r Memory B cells r Vaccination Additional supporting information may be found in the online version of this article at the publisher's web-site IntroductionThe influenza A virus surface proteins hemagglutinin (HA) and neuraminidase (NA) are highly susceptible to mutations that favor the selection of variants able to escape neutralizing antibodies. This phenomenon, known as antigenic drift, necessitates the proCorrespondence: Dr. Oretta Finco e-mail: oretta.finco@novartis.com duction of new influenza vaccines every year. More rarely, reassortment of genome segments from animal and human viruses might also occur, leading to the emergence of new viruses to which the human population is immunologically naïve (antigenic shift). These viruses, like avian H5N1, have the potential to cause pandemics [1][2][3].The immunological naivety of the human population to H5N1 is inferred by the low proportion of adults with hemagglutinationinhibiting antibodies to H5N1 virus [4]. In addition, clinical trials with H5N1 and H9N2 vaccines have shown that two doses of C 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.eji-journal.eu 642Francesca Buricchi et al. Eur. J. Immunol. 2013. 43: 641-648 adjuvanted vaccine are needed in order to increase neutralizing antibody titers to potentially protective levels [5][6][7][8][9][10]. In spite of this, it has been observed that vaccination with seasonal strains or natural influenza infection can transiently induce antibodies neutralizing the H5N1 virus [11,12]. Heterosubtypic monoclonal antibodies able to cross-react across group 1 or group 2 influenza virus subtypes have been isolated from MBCs and plasma cells of adults e...

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Section: Preexisting H1n1-igg Mbcs May Favor a Functional Response Tomentioning

confidence: 96%

Section: Pseudoparticle Neutralization Assaymentioning

confidence: 99%

Section: Preexisting H1n1-igg Mbcs May Favor a Functional Response Tomentioning

confidence: 99%

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Impact of preexisting memory to seasonal A/H1N1 influenza virus on the immune response following vaccination against avian A/H5N1 virus

et al. 2013

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“…The responses in senior adults nearly reached those levels. In a pseudotype neutralization assay, primary responses in young adults also were shown to be broadly reactive to clade 2.1 and 2.3.4 antigens [98].…”

Section: Pandemic Influenza Vaccines Containing Mf59: Avian Influenzamentioning

confidence: 99%

Emulsion-Based Adjuvants for Improved Influenza Vaccines

O’Hagan

Tsai

Reed

2010

Influenza Vaccines for the Future

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Emulsions have a long history of use as potent and effective adjuvants in humans for a range of vaccines, particularly for influenza. Although older mineral oil-and water-in-oil-based emulsion adjuvants did not have an overall safety and tolerability profile to allow them to be acceptable for widespread use, a newer generation of oil-in-water adjuvants has been recently developed, based on the use of the biodegradable oil squalene. These adjuvants have shown particular value in the development of new generation vaccines to offer enhanced protection against both seasonal and pandemic strains of influenza virus. The first oil-in-water emulsion adjuvant included in an approved flu vaccine was MF59, which was originally licensed in Europe in 1997 as an improved influenza vaccine for the elderly. In the very recent past, MF59 and related adjuvants have shown their value by offering the possibility of significant antigen dose reductions and higher potency products in the face of the H1N1 pandemic emergency and other pandemic threats. The recent H1N1 global problem allowed the opportunity for widespread use of emulsionbased adjuvants in a range of population groups in a number of countries, in which strict monitoring of safety was the norm. Importantly, this widespread use allowed the safety profile of squalene-based emulsion adjuvants to be further substantiated in large and diverse populations of humans, including young children and pregnant women. It is our confident prediction that the coming years will see wider use and further licensures for oil-in-water emulsion adjuvants, particularly for improved flu vaccines.

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“…According to the type of detection target, these methods could be categorized into, for example, virus isolation and identification, nucleic acid-based detection, antigen detection, and antibody detection [3][4][5][6][7][8]. Most of these methods need demanding conditions and professional operations, and some of the detection processes is time-consuming.…”

mentioning

confidence: 99%

Ultra Sensitive Detection of Influenza A Virus Based on Cdse/Zns Quantum Dots Immunoassay

Wu¹,

Mao²,

Liu³

et al. 2016

SOJB

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Open Access Research articlesevere outbreaks disease, and they can cause serious infections among people.To date, numerous analytical methods have being used to detect influenza A virus. According to the type of detection target, these methods could be categorized into, for example, virus isolation and identification, nucleic acid-based detection, antigen detection, and antibody detection [3][4][5][6][7][8]. Most of these methods need demanding conditions and professional operations, and some of the detection processes is time-consuming. For these reasons, some new biosensors based on nano materials such as gold nanoparticles, magnetic nano beads and quantum dots (QDs) have been coming to the forefront [9][10][11][12].Developing a sensitive, specific and fast biosensor for diagnosing the influenza virus at the early stages of infection is a challenge. Lateral flow immune assay method has been widely used in detection of infectious diseases with the advantages of rapid, easy to operate, and low cost [13]. However, traditional method such as colloidal gold lateral flow tests has limitations when high sensitivity is needed [14].QDs have been developed to replace colloidal gold owing to their excellent optical properties such as high fluorescence efficiency, wide range of excitation wavelength, narrow and symmetric emission spectra, and QDs based lateral flow tests have been proved higher sensitivity and reproducibility for detection of pathogen, characteristic protein and even nucleic acid [15][16][17][18].In this paper, we present an ultrasensitive lateral flow immunoassay (LFIA) for detecting influenza A virus. This immune sensor is based on site-specific covalent binding with the Fc end of influenza A virus antibodies to CdSe/ ZnS QDs. The carboxylfunctionalized CdSe/ ZnS QDs conjugated with antibodies via an amide bond often result in random links of the antibody structure in the QD conjugates and block certain antigenbinding sites. In order to obtain site-specific linking, we modified carboxyl-functionalized QDs with adipicdihydrazide (ADH) and oxidized the carbohydrate groups on the antibody's Fc region AbstractAn ultrasensitive lateral flow immunoassay system (LFIAS) was established for the detection of influenza A virus. In this LFIAS, hydrophilic dihydrazide-modified CdSe/ZnSquantom dots (QDs) were conjugated with specific antibodies and used as fluorescent labels, and a pair of matched anti-nucleoprotein of influenza A virus antibodies were used to form a sandwich immunoassay. The QDs were in conjugation with the fragment crystallizable region (Fc region) of specific influenza A virus antibodies through aldehydehydrazide covalent chemistry, conferring high sensitivity. The antibodies used for detection are specific for the most conserved and popular nucleoprotein of influenza A virus and ensure the accuracy and specificity. The QDs-LFIAS can analyze the nasal-pharyngeal swab samples through simple steps and get results within 15 min. Detection of nasal-pharyngeal swab samples makes it more rapid and conve...

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Pseudoparticle neutralization is a reliable assay to measure immunity and cross-reactivity to H5N1 influenza viruses

Cited by 61 publications

References 20 publications

Impact of preexisting memory to seasonal A/H1N1 influenza virus on the immune response following vaccination against avian A/H5N1 virus

Impact of preexisting memory to seasonal A/H1N1 influenza virus on the immune response following vaccination against avian A/H5N1 virus

Emulsion-Based Adjuvants for Improved Influenza Vaccines

Ultra Sensitive Detection of Influenza A Virus Based on Cdse/Zns Quantum Dots Immunoassay

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