Frontiers in Immunology (Dec 2020)
Modeling MyD88 Deficiency In Vitro Provides New Insights in Its Function
- Nils Craig-Mueller,
- Nils Craig-Mueller,
- Nils Craig-Mueller,
- Ruba Hammad,
- Ruba Hammad,
- Ruba Hammad,
- Roland Elling,
- Roland Elling,
- Roland Elling,
- Jamal Alzubi,
- Jamal Alzubi,
- Barbara Timm,
- Barbara Timm,
- Julia Kolter,
- Julia Kolter,
- Nele Knelangen,
- Nele Knelangen,
- Christien Bednarski,
- Christien Bednarski,
- Birgitta Gläser,
- Sandra Ammann,
- Sandra Ammann,
- Zoltán Ivics,
- Judith Fischer,
- Judith Fischer,
- Carsten Speckmann,
- Carsten Speckmann,
- Carsten Speckmann,
- Klaus Schwarz,
- Nico Lachmann,
- Nico Lachmann,
- Stephan Ehl,
- Stephan Ehl,
- Stephan Ehl,
- Thomas Moritz,
- Thomas Moritz,
- Philipp Henneke,
- Philipp Henneke,
- Philipp Henneke,
- Philipp Henneke,
- Toni Cathomen,
- Toni Cathomen,
- Toni Cathomen
Affiliations
- Nils Craig-Mueller
- Institute for Transfusion Medicine and Gene Therapy, Medical Center–University of Freiburg, Freiburg, Germany
- Nils Craig-Mueller
- Center for Chronic Immunodeficiency (CCI), Medical Center–University of Freiburg, Freiburg, Germany
- Nils Craig-Mueller
- MD Program, Faculty of Medicine, University of Freiburg, Freiburg, Germany
- Ruba Hammad
- Institute for Transfusion Medicine and Gene Therapy, Medical Center–University of Freiburg, Freiburg, Germany
- Ruba Hammad
- Center for Chronic Immunodeficiency (CCI), Medical Center–University of Freiburg, Freiburg, Germany
- Ruba Hammad
- PhD Program, Faculty of Biology, University of Freiburg, Freiburg, Germany
- Roland Elling
- Center for Chronic Immunodeficiency (CCI), Medical Center–University of Freiburg, Freiburg, Germany
- Roland Elling
- Institute for Immunodeficiency, Medical Center–University of Freiburg, Freiburg, Germany
- Roland Elling
- Center for Pediatrics and Adolescent Medicine, Medical Center–University of Freiburg, Freiburg, Germany
- Jamal Alzubi
- Institute for Transfusion Medicine and Gene Therapy, Medical Center–University of Freiburg, Freiburg, Germany
- Jamal Alzubi
- Center for Chronic Immunodeficiency (CCI), Medical Center–University of Freiburg, Freiburg, Germany
- Barbara Timm
- Institute for Transfusion Medicine and Gene Therapy, Medical Center–University of Freiburg, Freiburg, Germany
- Barbara Timm
- Center for Chronic Immunodeficiency (CCI), Medical Center–University of Freiburg, Freiburg, Germany
- Julia Kolter
- Center for Chronic Immunodeficiency (CCI), Medical Center–University of Freiburg, Freiburg, Germany
- Julia Kolter
- Institute for Immunodeficiency, Medical Center–University of Freiburg, Freiburg, Germany
- Nele Knelangen
- Center for Chronic Immunodeficiency (CCI), Medical Center–University of Freiburg, Freiburg, Germany
- Nele Knelangen
- Institute for Immunodeficiency, Medical Center–University of Freiburg, Freiburg, Germany
- Christien Bednarski
- Institute for Transfusion Medicine and Gene Therapy, Medical Center–University of Freiburg, Freiburg, Germany
- Christien Bednarski
- Center for Chronic Immunodeficiency (CCI), Medical Center–University of Freiburg, Freiburg, Germany
- Birgitta Gläser
- Institute of Human Genetics, Medical Center–University of Freiburg, Freiburg, Germany
- Sandra Ammann
- Center for Chronic Immunodeficiency (CCI), Medical Center–University of Freiburg, Freiburg, Germany
- Sandra Ammann
- Institute for Immunodeficiency, Medical Center–University of Freiburg, Freiburg, Germany
- Zoltán Ivics
- Division of Medical Biotechnology, Paul-Ehrlich Institute, Langen, Germany
- Judith Fischer
- Institute of Human Genetics, Medical Center–University of Freiburg, Freiburg, Germany
- Judith Fischer
- Faculty of Medicine, University of Freiburg, Freiburg, Germany
- Carsten Speckmann
- Center for Chronic Immunodeficiency (CCI), Medical Center–University of Freiburg, Freiburg, Germany
- Carsten Speckmann
- Center for Pediatrics and Adolescent Medicine, Medical Center–University of Freiburg, Freiburg, Germany
- Carsten Speckmann
- Faculty of Medicine, University of Freiburg, Freiburg, Germany
- Klaus Schwarz
- 0Institute for Clinical Transfusion Medicine and Immunogenetics Ulm, German Red Cross Blood Service Baden-Württemberg–Hessen, and Institute for Transfusion Medicine, University of Ulm, Ulm, Germany
- Nico Lachmann
- 1Institute of Experimental Hematology, Hannover Medical School, Hannover, Germany
- Nico Lachmann
- 2REBIRTH Cluster for Regenerative and Translational Medicine, Hannover, Germany
- Stephan Ehl
- Center for Chronic Immunodeficiency (CCI), Medical Center–University of Freiburg, Freiburg, Germany
- Stephan Ehl
- Institute for Immunodeficiency, Medical Center–University of Freiburg, Freiburg, Germany
- Stephan Ehl
- Faculty of Medicine, University of Freiburg, Freiburg, Germany
- Thomas Moritz
- 1Institute of Experimental Hematology, Hannover Medical School, Hannover, Germany
- Thomas Moritz
- 2REBIRTH Cluster for Regenerative and Translational Medicine, Hannover, Germany
- Philipp Henneke
- Center for Chronic Immunodeficiency (CCI), Medical Center–University of Freiburg, Freiburg, Germany
- Philipp Henneke
- Institute for Immunodeficiency, Medical Center–University of Freiburg, Freiburg, Germany
- Philipp Henneke
- Center for Pediatrics and Adolescent Medicine, Medical Center–University of Freiburg, Freiburg, Germany
- Philipp Henneke
- Faculty of Medicine, University of Freiburg, Freiburg, Germany
- Toni Cathomen
- Institute for Transfusion Medicine and Gene Therapy, Medical Center–University of Freiburg, Freiburg, Germany
- Toni Cathomen
- Center for Chronic Immunodeficiency (CCI), Medical Center–University of Freiburg, Freiburg, Germany
- Toni Cathomen
- Faculty of Medicine, University of Freiburg, Freiburg, Germany
- DOI
- https://doi.org/10.3389/fimmu.2020.608802
- Journal volume & issue
-
Vol. 11
Abstract
Inherited defects in MyD88 and IRAK4, two regulators in Toll-like receptor (TLR) signaling, are clinically highly relevant, but still incompletely understood. MyD88- and IRAK4-deficient patients are exceedingly susceptible to a narrow spectrum of pathogens, with ∼50% lethality in the first years of life. To better understand the underlying molecular and cellular characteristics that determine disease progression, we aimed at modeling the cellular response to pathogens in vitro. To this end, we determined the immunophenotype of monocytes and macrophages derived from MyD88- and IRAK4-deficient patients. We recognized that macrophages derived from both patients were particularly poorly activated by streptococci, indicating that both signaling intermediates are essential for the immune response to facultative pathogens. To characterize this defect in more detail, we generated induced pluripotent stem cells (iPSCs) of fibroblasts derived from an MyD88-deficient patient. The underlying genetic defect was corrected using Sleeping Beauty transposon vectors encoding either the long (L) or the short (S) MYD88 isoform, respectively. Macrophages derived from these iPSC lines (iMacs) expressed typical macrophage markers, stably produced either MyD88 isoform, and showed robust phagocytic activity. Notably, iMacs expressing MyD88-L, but not MyD88-S, exhibited similar responses to external stimuli, including cytokine release patterns, as compared to genetically normal iMacs. Thus, the two MyD88 isoforms assume distinct functions in signaling. In conclusion, iPSC technology, in combination with efficient myeloid differentiation protocols, provides a valuable and inexhaustible source of macrophages, which can be used for disease modeling. Moreover, iPSC-derived macrophages may eventually aid in stabilizing MyD88-deficient patients during pyogenic infections.
Keywords