Dataset Information

Impaired IFN?-Signaling and Mycobacterial Clearance in IFN?R1-Deficient Human iPSC-Derived Macrophages.

ABSTRACT: Mendelian susceptibility to mycobacterial disease (MSMD) is caused by inborn errors of interferon gamma (IFN?) immunity and is characterized by severe infections by weakly virulent mycobacteria. Although IFN? is the macrophage-activating factor, macrophages from these patients have never been studied. We demonstrate the generation of heterozygous and compound heterozygous (iMSMD-cohet) induced pluripotent stem cells (iPSCs) from a single chimeric patient, who suffered from complete autosomal recessive IFN?R1 deficiency and received bone-marrow transplantation. Loss of IFN?R1 expression had no influence on the macrophage differentiation potential of patient-specific iPSCs. In contrast, lack of IFN?R1 in iMSMD-cohet macrophages abolished IFN?-dependent phosphorylation of STAT1 and induction of IFN?-downstream targets such as IRF-1, SOCS-3, and IDO. As a consequence, iMSMD-cohet macrophages show impaired upregulation of HLA-DR and reduced intracellular killing of Bacillus Calmette-Guérin. We provide a disease-modeling platform that might be suited to investigate novel treatment options for MSMD and to gain insights into IFN? signaling in macrophages.

SUBMITTER: Neehus AL

PROVIDER: S-EPMC5768914 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature

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Impaired IFNγ-Signaling and Mycobacterial Clearance in IFNγR1-Deficient Human iPSC-Derived Macrophages.

Neehus Anna-Lena AL Lam Jenny J Haake Kathrin K Merkert Sylvia S Schmidt Nico N Mucci Adele A Ackermann Mania M Schubert Madline M Happle Christine C Kühnel Mark Philipp MP Blank Patrick P Philipp Friederike F Goethe Ralph R Jonigk Danny D Martin Ulrich U Kalinke Ulrich U Baumann Ulrich U Schambach Axel A Roesler Joachim J Lachmann Nico N

Stem cell reports 20171214 1

Mendelian susceptibility to mycobacterial disease (MSMD) is caused by inborn errors of interferon gamma (IFNγ) immunity and is characterized by severe infections by weakly virulent mycobacteria. Although IFNγ is the macrophage-activating factor, macrophages from these patients have never been studied. We demonstrate the generation of heterozygous and compound heterozygous (iMSMD-cohet) induced pluripotent stem cells (iPSCs) from a single chimeric patient, who suffered from complete autosomal rec ...[more]

PMID: 29249666

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Project description:Despite the presence of significant levels of systemic Interferon gamma (IFNγ), the host protective cytokine, Kala-azar patients display high parasite load with downregulated IFNγ signaling in Leishmania donovani (LD) infected macrophages (LD-MØs); the cause of such aberrant phenomenon is unknown. Here we reveal for the first time the mechanistic basis of impaired IFNγ signaling in parasitized murine macrophages. Our study clearly shows that in LD-MØs IFNγ receptor (IFNγR) expression and their ligand-affinity remained unaltered. The intracellular parasites did not pose any generalized defect in LD-MØs as IL-10 mediated signal transducer and activator of transcription 3 (STAT3) phosphorylation remained unaltered with respect to normal. Previously, we showed that LD-MØs are more fluid than normal MØs due to quenching of membrane cholesterol. The decreased rigidity in LD-MØs was not due to parasite derived lipophosphoglycan (LPG) because purified LPG failed to alter fluidity in normal MØs. IFNγR subunit 1 (IFNγR1) and subunit 2 (IFNγR2) colocalize in raft upon IFNγ stimulation of normal MØs, but this was absent in LD-MØs. Oddly enough, such association of IFNγR1 and IFNγR2 could be restored upon liposomal delivery of cholesterol as evident from the fluorescence resonance energy transfer (FRET) experiment and co-immunoprecipitation studies. Furthermore, liposomal cholesterol treatment together with IFNγ allowed reassociation of signaling assembly (phospho-JAK1, JAK2 and STAT1) in LD-MØs, appropriate signaling, and subsequent parasite killing. This effect was cholesterol specific because cholesterol analogue 4-cholestene-3-one failed to restore the response. The presence of cholesterol binding motifs [(L/V)-X(1-5)-Y-X(1-5)-(R/K)] in the transmembrane domain of IFNγR1 was also noted. The interaction of peptides representing this motif of IFNγR1 was studied with cholesterol-liposome and analogue-liposome with difference of two orders of magnitude in respective affinity (K(D): 4.27×10(-9) M versus 2.69×10(-7) M). These observations reinforce the importance of cholesterol in the regulation of function of IFNγR1 proteins. This study clearly demonstrates that during its intracellular life-cycle LD perturbs IFNγR1 and IFNγR2 assembly and subsequent ligand driven signaling by quenching MØ membrane cholesterol.

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Dataset Information

Impaired IFN?-Signaling and Mycobacterial Clearance in IFN?R1-Deficient Human iPSC-Derived Macrophages.

Publications

Impaired IFNγ-Signaling and Mycobacterial Clearance in IFNγR1-Deficient Human iPSC-Derived Macrophages.

Similar Datasets

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