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Overview of Serological Techniques for Influenza Vaccine Evaluation: Past, Present and Future.

Trombetta CM, Perini D, Mather S, Temperton N, Montomoli E - Vaccines (Basel) (2014)

Bottom Line: HI and SRH are established and reproducible techniques, whereas VN is more demanding.Every new influenza vaccine needs to fulfil the strict criteria issued by the European Medicines Agency (EMA) in order to be licensed.These criteria currently apply exclusively to SRH and HI assays and refer to two different target groups-healthy adults and the elderly, but other vaccine recipient age groups have not been considered (i.e., children).

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Developmental Medicine, University of Siena, Via Aldo Moro, 53100 Siena, Italy. trombetta@unisi.it.

ABSTRACT
Serological techniques commonly used to quantify influenza-specific antibodies include the Haemagglutination Inhibition (HI), Single Radial Haemolysis (SRH) and Virus Neutralization (VN) assays. HI and SRH are established and reproducible techniques, whereas VN is more demanding. Every new influenza vaccine needs to fulfil the strict criteria issued by the European Medicines Agency (EMA) in order to be licensed. These criteria currently apply exclusively to SRH and HI assays and refer to two different target groups-healthy adults and the elderly, but other vaccine recipient age groups have not been considered (i.e., children). The purpose of this timely review is to highlight the current scenario on correlates of protection concerning influenza vaccines and underline the need to revise the criteria and assays currently in use. In addition to SRH and HI assays, the technical advantages provided by other techniques such as the VN assay, pseudotype-based neutralization assay, neuraminidase and cell-mediated immunity assays need to be considered and regulated via EMA criteria, considering the many significant advantages that they could offer for the development of effective vaccines.

No MeSH data available.


Related in: MedlinePlus

Representation of the production and assay of influenza HA pseudotyped retroviruses [72].
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vaccines-02-00707-f003: Representation of the production and assay of influenza HA pseudotyped retroviruses [72].

Mentions: A pseudotype virus has the “core” of one virus (e.g., a retrovirus) and the outer “envelope” protein(s) of another (e.g., the HA/NA of influenza virus). The core virus has deletions in its genome making it replication-deficient (allowing it to be used under BSL1/2 containment), and harbours a reporter transgene (e.g., luciferase or Green Fluorescent Protein (GFP)). The envelope glycoprotein permits entry into susceptible target assay cells (e.g., Human Embryonic Kidney 293 T cells (HEK293T)/MDCK) via interaction with sialic acid. During target cell transduction, the pseudotype virus genome becomes integrated into the cell genome, resulting in reporter gene expression. Thus, the number of transduced cells can be accurately quantified (via a luminometer, fluorescent microscope or Fluorescence-Activated Cell Sorting (FACS)) and the subsequent inhibitory effects of functional antibodies (directed against the HA1 head and the HA2 stalk) in serum determined [67,68,69]. Figure 3 below shows a schematic representation of the production and assay of influenza HA pseudotyped retroviruses. Pseudotype production of Highly Pathogenic Avian Influenza (HPAI) strains (which have a polybasic cleavage site in the HA) routinely involves the transfection of three plasmids into 293T producer cells: retroviral gag-pol plasmid, HA-expressing plasmid, and retroviral vector plasmid incorporating the reporter gene. Additionally, for the production of Low Pathogenic Avian Influenza (LPAI) strains (single arginine at cleavage site), an additional plasmid expressing a protease (either Transmembrane Protease Serine 2 ((TMPRSS2) or Human Airway Trypsin-like Protease (HAT)) is necessary [70,71].


Overview of Serological Techniques for Influenza Vaccine Evaluation: Past, Present and Future.

Trombetta CM, Perini D, Mather S, Temperton N, Montomoli E - Vaccines (Basel) (2014)

Representation of the production and assay of influenza HA pseudotyped retroviruses [72].
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494249&req=5

vaccines-02-00707-f003: Representation of the production and assay of influenza HA pseudotyped retroviruses [72].
Mentions: A pseudotype virus has the “core” of one virus (e.g., a retrovirus) and the outer “envelope” protein(s) of another (e.g., the HA/NA of influenza virus). The core virus has deletions in its genome making it replication-deficient (allowing it to be used under BSL1/2 containment), and harbours a reporter transgene (e.g., luciferase or Green Fluorescent Protein (GFP)). The envelope glycoprotein permits entry into susceptible target assay cells (e.g., Human Embryonic Kidney 293 T cells (HEK293T)/MDCK) via interaction with sialic acid. During target cell transduction, the pseudotype virus genome becomes integrated into the cell genome, resulting in reporter gene expression. Thus, the number of transduced cells can be accurately quantified (via a luminometer, fluorescent microscope or Fluorescence-Activated Cell Sorting (FACS)) and the subsequent inhibitory effects of functional antibodies (directed against the HA1 head and the HA2 stalk) in serum determined [67,68,69]. Figure 3 below shows a schematic representation of the production and assay of influenza HA pseudotyped retroviruses. Pseudotype production of Highly Pathogenic Avian Influenza (HPAI) strains (which have a polybasic cleavage site in the HA) routinely involves the transfection of three plasmids into 293T producer cells: retroviral gag-pol plasmid, HA-expressing plasmid, and retroviral vector plasmid incorporating the reporter gene. Additionally, for the production of Low Pathogenic Avian Influenza (LPAI) strains (single arginine at cleavage site), an additional plasmid expressing a protease (either Transmembrane Protease Serine 2 ((TMPRSS2) or Human Airway Trypsin-like Protease (HAT)) is necessary [70,71].

Bottom Line: HI and SRH are established and reproducible techniques, whereas VN is more demanding.Every new influenza vaccine needs to fulfil the strict criteria issued by the European Medicines Agency (EMA) in order to be licensed.These criteria currently apply exclusively to SRH and HI assays and refer to two different target groups-healthy adults and the elderly, but other vaccine recipient age groups have not been considered (i.e., children).

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Developmental Medicine, University of Siena, Via Aldo Moro, 53100 Siena, Italy. trombetta@unisi.it.

ABSTRACT
Serological techniques commonly used to quantify influenza-specific antibodies include the Haemagglutination Inhibition (HI), Single Radial Haemolysis (SRH) and Virus Neutralization (VN) assays. HI and SRH are established and reproducible techniques, whereas VN is more demanding. Every new influenza vaccine needs to fulfil the strict criteria issued by the European Medicines Agency (EMA) in order to be licensed. These criteria currently apply exclusively to SRH and HI assays and refer to two different target groups-healthy adults and the elderly, but other vaccine recipient age groups have not been considered (i.e., children). The purpose of this timely review is to highlight the current scenario on correlates of protection concerning influenza vaccines and underline the need to revise the criteria and assays currently in use. In addition to SRH and HI assays, the technical advantages provided by other techniques such as the VN assay, pseudotype-based neutralization assay, neuraminidase and cell-mediated immunity assays need to be considered and regulated via EMA criteria, considering the many significant advantages that they could offer for the development of effective vaccines.

No MeSH data available.


Related in: MedlinePlus