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A significant percentage of patients are also Multi Drug Resistant TB(MDR-TB) suspects. KENJIRO SHIMA1, TAKURO SAKAGAMI1, YOSHINARI TANABE1, NOBUMASA AOKI1, HIROSHI MORO1, TOSHIYUKI KOYA1, TAKASHI HASEGAWA2, EI-ICHI SUZUKI2, ICHIEI NARITA1 1Division of Respiratory Medicine and Infectious Disease, Niigata University, Niigata, Japan, 2Department of General Medicine, Niigata University Medical and Dental Hospital, Niigata, Japan Rationale?All of exposed subjects with non-tubelculouse mycobacterium (NTM) species do not develop active infection, which likely reflects underlying host susceptibility factors. Recent reports have shown that anti Interferon-Gamma (IFN-gamma) neutralizing autoantibodies (IFN-gamma DDR1 Ab) are associated with disseminated NTM patients without known evidence of immunodeficiency. The purpose of this study is to establish the screening method if subjects have IFN-gamma Ab. Here, we report a novel bioassay to evaluate IFN-gamma signaling in whole blood and human T-cell line with patients plasma based on IFN-gamma stimulated, IFN-gamma receptor-mediated phosphorylation of STAT1, which is named the STAT1 phosphorylation index (STAT1-PI). Method?Blood was obtained from patients with disseminated NTM, pulmonary NTM and healthy controls. To evaluate neutralizing capacity to IFN-gamma, STAT-1 phosphorylation Veliparib mw in leukocyte and T-cell line after stimulation with various concentrations of exogenous IFN-gamma (ranging from 1 to 1000?ng/ml) was evaluated by using flow cytometer. The strength of phosphorylation was described as STAT1-PI which was calculated as the mean fluorescence of cells primed with assigned concentration of IFN-gamma minus that of unprimed cells divided by that of unprimed cells multiplied by 100. Antigen capture assay was performed to measure the relative titer of Ig-G fraction of IFN-gamma Ab. Result?The STAT1-PI increased in proportion to IFN-gamma concentration reaching a maximum value near 1000?ng/ml and to time reaching in 30 minutes after stimulation. STAT1-PI with plasma from disseminated NTM patients was inhibited, while it from other disease and healthy subjects were not (n?=?6: 7.3 plus or minus 27.8, n?=?3: 424.8 plus or minus 21.9, n?=?6: 243.4 plus or minus 133.9, p?Autophagy activator concentration of IFN-gamma Ab(mean: 493 E.U.). Conclusion?The STAT1-PI is simple, useful flow cytometry-based method to measure IFN-gamma signaling in the evaluation of disseminated NTM patients suspected of having IFN-gamma Ab. JAECHUN LEE, MIOK KIM, JONG HOO LEE, GIL MYEONG SEONG Department of Internal Medicine, Jeju National University School of Medicine, Jeju, Korea Backgrounds?The bacteriologic diagnosis in pulmonary tuberculosis is confirmative, even though successful rates in self-expectorated sputum are limited.