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. Author manuscript; available in PMC: 2023 Mar 23.
Published in final edited form as: Nat Biomed Eng. 2022 Aug 19;6(8):992–1003. doi: 10.1038/s41551-022-00925-y

Fig. 5 |. MIPSA detects known and novel neutralizing interferon autoantibodies.

Fig. 5 |

a-c, Scatterplots highlighting reactive interferon UCIs for three severe COVID-19 patients. d, Summary of interferon reactivity detected in 5 of 55 individuals with severe COVID-19. Hits fold-change values (colour of cell) and the number of reactive UCIs (number in cell) are provided. e, f, Recombinant interferon alpha 2 (IFN-α2) or interferon lambda 3 (IFN-λ3) neutralizing activity of the same patients shown in d. Plasma were pre-incubated with 100 U/ml of IFN-α2 or 1 ng/ml of IFN-λ3 prior to incubation with A549 cells. Fold changes of the interferon stimulated gene, MX1, were calculated by RT-qPCR relative to unstimulated cells (n = 3 for each of the 5 patient samples). For e, samples labelled HC, P1, P2, P3, P4, P5 have mean and standard deviation values of 977.7 +/−253.7, 1.39 +/−0.2, 3.1 +/−1.8, 1.4 +/−0.5, 32 +/−7.2, 741.1 +/−121, respectively. For f, samples labelled HC, P1, P2, P3, P4, P5 have mean and standard deviation values of 64.5 +/−26.8, 28.2 +/−12.1, 1.6 +/− 1.0, 20.3 +/−1.4, 52.4 +/− 5.1, 2.3 +/−2.0, respectively. GAPDH was used as a housekeeping control gene for normalization. Red bars indicate which samples were found to contain the corresponding anti-interferon antibodies using MIPSA. g, PhIP-Seq analysis of interferon autoantibodies in the 5 patients of d (row and column orders maintained). Hits fold-change values (colour of cell) and the number of reactive peptides (number in cell) are provided. h, Epitopefindr analysis of the PhIP-Seq reactive type I interferon 90-aa peptides.