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Penicillium marneffei infection and impaired IFN-γ immunity in humans with autosomal- dominant gain-of-phosphorylation STAT1 mutations  Pamela P.W. Lee,

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Presentation on theme: "Penicillium marneffei infection and impaired IFN-γ immunity in humans with autosomal- dominant gain-of-phosphorylation STAT1 mutations  Pamela P.W. Lee,"— Presentation transcript:

1 Penicillium marneffei infection and impaired IFN-γ immunity in humans with autosomal- dominant gain-of-phosphorylation STAT1 mutations  Pamela P.W. Lee, MBBS, Huawei Mao, PhD, Wanling Yang, PhD, Koon-Wing Chan, MSc, Marco H.K. Ho, MBBS, Tsz-Leung Lee, MBBS, Jasper F.W. Chan, MBBS, Patrick C.Y. Woo, MD, Wenwei Tu, PhD, Yu-Lung Lau, MD  Journal of Allergy and Clinical Immunology  Volume 133, Issue 3, Pages e5 (March 2014) DOI: /j.jaci Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Gain-of-phosphorylation STAT1 mutations impaired IFN-γ and IL-17 responses. PBMCs were stimulated with IFN-α (A) or IFN-γ (B) and analyzed for intracellular pSTAT1 expression by gating on lymphocytes. The increase in %pSTAT1+ population in stimulated cells relative to unstimulated cells was calculated. Representative histograms are shown for patient 1 and a normal control. C, PBMCs from patient 1 were stimulated by IFN-γ followed by treatment with staurosporine for 30 or 60 minutes. The percentage of intracellular pSTAT1 expression and mean fluorescence intensity (MFI) were determined in monocytes by flow cytometry. D, PBMCs were stimulated with phorbol 12-myristate 13-acetate plus ionomycin, and intracellular expression of IFN-γ and IL-17A in CD3+ T cells was analyzed by using flow cytometry. E, PBMCs were cocultured with Candida albicans (multiplicity of infection [MOI] of 5) or Penicillium marneffei conidia (MOI of 1) for 48 hours, and IFN-γ in the supernatant was quantified. P, Patient. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig E1 Schematic model of STAT1 mutations. Mutations of patient 1, patient 2, and patient 3 are indicated by asterisks, which shows all STAT1 mutations reported in the literature. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E2 STAT1 phosphorylation induced by IFN-α and IFN-γ. PBMCs of patients and controls were stimulated with IFN-α or IFN-γ for 20 minutes. The percentage of intracellular phosphorylated STAT1 (pSTAT1) expression in lymphocyte was analyzed by using flow cytometry. Representative histograms are presented, and the numbers in the right upper corner indicate the mean fluorescence intensity. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E3 Cytokine expression induced by Candia albicans and Penicillium marneffei (PM) stimulation. Individual PBMCs were cocultured with C albicans (multiplicity of infection [MOI] of 5) or P marneffei conidia (MOI of 1) for 48 hours. The cytokine production in the supernatant was examined by FlowCytomix according to manufacturer's instruction. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E4 Alignment of sequencing reads around the mutation in STAT1. This was performed by Integrative Genomics Viewer. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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