Influenza-induced IFN-γ production in vitro

Heparinised whole blood was diluted 1:10 with serum-free RPMI 1640 medium and incubated with either A/California/07/2009 (H1N1) or A/Perth/16/2009 (H3N2) allantoic fluids (1.6 × 10⁶ PFU). Phytohemagglutinin PHA-M (Sigma-Aldrich, Dorset, UK) was used as a positive control and egg allantoic fluid was used as a negative control. Blood was stimulated for 4 days at 37 °C, after which plasma supernatants were collected and cryopreserved at −80 °C. The Ferret IFN-γ ELISA Development Kit (ALP) (Mabtech, Nacka, Sweden) was used to determine the quantity of IFN-ɣ secreted by cells in the blood as described^{18 (link)}.

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Marriott A.C., Gooch K.E., Brown P.J., Ryan K.A., Jones N.J., Merredew N., Wiblin N., Dibben O., Bright H., Hallis B., Whittaker C.J, & Carroll M.W. (2021). Severity of heterosubtypic influenza virus infection in ferrets is reduced by live attenuated influenza vaccine. NPJ Vaccines, 6, 43.

Publication 2021

Phytohemagglutinin PHA-M Ferret IFN-γ ELISA Development Kit (ALP)

Allantoic Cells Elisa Ferret Ifn γ In blood Plasma Serum

Corresponding Organization : Public Health England

Other organizations : AstraZeneca (United Kingdom)

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Variable analysis

independent variables

A/California/07/2009 (H1N1) allantoic fluids (1.6 × 10^6 PFU)
A/Perth/16/2009 (H3N2) allantoic fluids (1.6 × 10^6 PFU)

dependent variables

Quantity of IFN-ɣ secreted by cells in the blood

control variables

Heparinised whole blood diluted 1:10 with serum-free RPMI 1640 medium
Incubation time of 4 days at 37 °C

positive control

Phytohemagglutinin PHA-M (Sigma-Aldrich, Dorset, UK)

negative control

Egg allantoic fluid

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