1. Immunology and Inflammation
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  1. Christina Hertel
  2. Dmytro Fishman
  3. Anna Lorenc
  4. Annamari Ranki
  5. Kai Krohn
  6. Pärt Peterson
  7. Kai Kisand  Is a corresponding author
  8. Adrian Hayday  Is a corresponding author
  1. ImmunoQure AG, Germany
  2. University of Tartu, Estonia
  3. King’s College London, United Kingdom
  4. University of Helsinki, Finland
  5. Francis Crick Institute, United Kingdom
Cite this article as: eLife 2019;8:e45826 doi: 10.7554/eLife.45826
1 figure, 2 tables, 1 data set and 1 additional file

Figures

The comparison of two different strategies to measure IFNα neutralizing capacity of APECED/APS1 serum samples.

In panel (A), the same reporter cell assay (HEK-Blue IFN-α/β cells from InvivoGen) has been applied as in Meyer et al. (2016) but at a single high serum concentration (ns: not significant). (B) Representative fitted dose-response curves that were used in Meyer et al. (2016) to calculate IC50 values for each serum sample. Individual curves are represented with dotted lines and those for grouped values in solid lines (mean ± SEM). (C) IC50 values (expressed as the dilution of serum sufficient to neutralize 50% of IFNα2 activity [12.5 U/ml]) that were calculated from individual and grouped curves shown in panel B. APECED/APS1 patients with Type 1 diabetes (T1D) are depicted in red and APECED/APS1 patients with GAD65 autoantibodies (GADA) but without T1D are in blue. (D) Neutralization of IFNα2 activity (10 000 U/ml) required to induce pSTAT1 was tested with different dilutions of sera from GAD seropositive patients with and without T1D. 2-way ANOVA was used to calculate P-values [ns – not significant, **p≤0.01, ***p≤0.001, ****p≤0.0001].

https://doi.org/10.7554/eLife.45826.002

Tables

Table 1
Testis- and cancer- associated non-cytokine targets screened by LIPS.
https://doi.org/10.7554/eLife.45826.004
TargetLIPS result
SPAG8pos
SPANXDpos
TEX264pos
CT45A3pos
GAPDHSpos
SPAG16pos
PDILTpos
GAGE1pos
SPATA7pos
GAGE7pos
CAPNS1pos
KCNIP2pos
POMZP3pos
MAGEA4pos
RPL12pos
MKNK2pos
S100A7Apos
MAGEA3pos
MAGEB1pos
MAGEB2pos
MAGEA10pos
LCN1pos
FGF12pos
HMGB1pos
TSPY2neg
MORN2neg
CRYGDneg
GNG4neg
RSU1neg
PAGE1neg
PAGE2neg
Key resources table
Reagent type or resourceDesignationSourceIdentifiers
Cell lineHuman HEK293 cells - Type I IFNs reporter cellsInvioGencat # hkb-ifnab
AntibodyAlexa Fluor 647 conjugated anti-STAT1 (pY701), mouse IgG2aBD Biosciencescat # 562070
Recombinant proteinrecombinant human IFNα2aMiltenyi Biotechcat # 130-093-873

Data availability

All data generated or analysed in this study are included in the manuscript and supporting files. ProtoArray data have been deposited at Array Express (E-MTAB-5369).

The following previously published data sets were used
  1. 1
    ArrayExpress
    1. Fishman Dmytro
    2. Peterson Pärt
    (2017)
    ID E-MTAB-5369. Protein microarray (Protoarray, Invitrogen) screening with APECED patient, healthy relative and healthy control serum samples.

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