Quantifying Anti-IFN-γ Autoantibodies

The method has been published^{7 (link),14 (link),20 (link),21 (link)}. Heparinized plasma from patients was serially diluted and incubated with recombinant human IFN-γ (BD Biosciences) at a final concentration of 300 pg/ml for 1 h at 37 °C. The level of unbound IFN-γ in the pre-incubation mixture was determined by a human IFN-γ ELISA kit (BD OptEIA: BD Biosciences), following the manufacturer’s instructions. The titer of the anti-human-IFN-γ autoantibody was determined in the highest dilution with >50% inhibition (<150 pg/ml of IFN-γ was detected).

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Nithichanon A., Chetchotisakd P., Matsumura T., Takahashi Y., Ato M., Sakagami T, & Lertmemongkolchai G. (2020). Diagnosis of NTM active infection in lymphadenopathy patients with anti-interferon-gamma auto-antibody using inhibitory ELISA vs. indirect ELISA. Scientific Reports, 10, 8968.

Publication 2020

recombinant human IFN-γ BD OptEIA

Autoantibody Dilution Elisa Human Ifn γ Inhibition Patients Plasma

Corresponding Organization :

Other organizations : Khon Kaen University, National Institute of Infectious Diseases, Kumamoto University Hospital

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Variable analysis

independent variables

Dilution of heparinized plasma from patients

dependent variables

Level of unbound IFN-γ in the pre-incubation mixture
Titer of the anti-human-IFN-γ autoantibody

control variables

Concentration of recombinant human IFN-γ (300 pg/ml)
Incubation time (1 h)
Incubation temperature (37 °C)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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