Evaluation of ability of anti-cytokine autoantibodies to prevent their respective cytokine signaling in vitro
| Autoantibody . | Control plasma . | Patients without detectable autoantibodies . | Number of patients with autoantibody . | Patients with neutralizing autoantibody . | Patients with nonneutralizing autoantibodies . | Comments on neutralization assays and results . |
|---|---|---|---|---|---|---|
| IFN-α | All nonneutralizing | All nonneutralizing | 10 | 10 | 0 | IFN-α1 used for LIPS but IFN-α2b used for functional assays; 86% similarity between these IFN-α species |
| Plasma prevented IFN-α–induced pSTAT-1 and pSTAT-4 production by flow cytometry and immunoblot, and production of IFN-α–inducible gene expression by qRT-PCR (supplemental Figures 1 and 2C) | ||||||
| IFN-β | All nonneutralizing | All nonneutralizing | 4 | 3 (patients 8, 9, and 11) | 1 (patient 2) | Plasma prevented IFN-β–induced pSTAT-1 production by flow cytometry and immunoblot and production of IFN-β–inducible gene expression by qRT-PCR (supplemental Figure 2) |
| IL-12p35 and IL-12p40 | All nonneutralizing | Plasma from 2 patients prevented IL-12–induced IFN-γ production and detection of IL-12 (supplemental Figure 3B-C) | Anti–IL-12p35 (1); anti–IL-12p40 (3); anti–IL-12p35 and p40 (6) | 10 | 0 | All 10 plasmas prevented IL-12–induced IFN-γ production and detection of IL-12. Plasma from 4 of 6 patients with antibodies against both IL-12p35 and p40 prevented IL-12–induced pSTAT-4 production by flow cytometry (supplemental Figure 3) |
| IL-1α | All nonneutralizing | All nonneutralizing | 2 | 1 (patient 1) | 1 (patient 7) | (supplemental Figure 4A-B) |
| IL-17 | All nonneutralizing | All nonneutralizing | 3 | 2 (patients 1 and 2) | 1 (patient 13) | (supplemental Figure 4C) |
| TNF-α | All nonneutralizing | All nonneutralizing | 1 | 0 | 1 (patient 8) | Anti-TNF-α autoantibodies in patient plasma did not prevent detection of TNF-α produced by PBMCs by luminex-based cytokine determination (data not shown) |
| Autoantibody . | Control plasma . | Patients without detectable autoantibodies . | Number of patients with autoantibody . | Patients with neutralizing autoantibody . | Patients with nonneutralizing autoantibodies . | Comments on neutralization assays and results . |
|---|---|---|---|---|---|---|
| IFN-α | All nonneutralizing | All nonneutralizing | 10 | 10 | 0 | IFN-α1 used for LIPS but IFN-α2b used for functional assays; 86% similarity between these IFN-α species |
| Plasma prevented IFN-α–induced pSTAT-1 and pSTAT-4 production by flow cytometry and immunoblot, and production of IFN-α–inducible gene expression by qRT-PCR (supplemental Figures 1 and 2C) | ||||||
| IFN-β | All nonneutralizing | All nonneutralizing | 4 | 3 (patients 8, 9, and 11) | 1 (patient 2) | Plasma prevented IFN-β–induced pSTAT-1 production by flow cytometry and immunoblot and production of IFN-β–inducible gene expression by qRT-PCR (supplemental Figure 2) |
| IL-12p35 and IL-12p40 | All nonneutralizing | Plasma from 2 patients prevented IL-12–induced IFN-γ production and detection of IL-12 (supplemental Figure 3B-C) | Anti–IL-12p35 (1); anti–IL-12p40 (3); anti–IL-12p35 and p40 (6) | 10 | 0 | All 10 plasmas prevented IL-12–induced IFN-γ production and detection of IL-12. Plasma from 4 of 6 patients with antibodies against both IL-12p35 and p40 prevented IL-12–induced pSTAT-4 production by flow cytometry (supplemental Figure 3) |
| IL-1α | All nonneutralizing | All nonneutralizing | 2 | 1 (patient 1) | 1 (patient 7) | (supplemental Figure 4A-B) |
| IL-17 | All nonneutralizing | All nonneutralizing | 3 | 2 (patients 1 and 2) | 1 (patient 13) | (supplemental Figure 4C) |
| TNF-α | All nonneutralizing | All nonneutralizing | 1 | 0 | 1 (patient 8) | Anti-TNF-α autoantibodies in patient plasma did not prevent detection of TNF-α produced by PBMCs by luminex-based cytokine determination (data not shown) |