Genetic Deficiency and Biochemical Inhibition of ITK Affect Human Th17, Treg, and Innate Lymphoid Cells

• DOI: 10.1007/s10875-019-00632-5
• Publication Date: 2019-04-25T22:04:13Z
• Authors: Ahmet Eken; Murat Cansever; Ido Somekh; Yoko Mizoguchi; Natalia Zietara; Fatma Zehra Okus; Serife Erdem; Halit Canatan; Sefika Akyol; Alper Ozcan; Musa Karakukcu; Sebastian Hollizeck; Meino Rohlfs; Ekrem Unal; Christoph Klein; Turkan Patiroglu
• Keywords: 0301 basic medicine; DNA Mutational Analysis; High-Throughput Nucleotide Sequencing; Apoptosis; Forkhead Transcription Factors; Protein-Tyrosine Kinases; Flow Cytometry; Immunity, Innate; Pedigree; 3. Good health; Consanguinity; Mice; 03 medical and health sciences; T-Lymphocyte Subsets; Child, Preschool; Animals; Cytokines; Humans; Female; Genetic Predisposition to Disease; Biomarkers; Genetic Association Studies; Cell Proliferation

Interleukin-2-inducible T cell kinase (ITK) is an important mediator of T cell receptor signaling. Loss of function mutations in ITK results in hypogammaglobulinemia and CD4+ T cell loss in humans, and the patients often present with EBV-associated B cell lymphoproliferative syndrome. Itk-deficient mice show loss of T cell naivety, impaired cytolytic activity of CD8+ T cells, and defects in CD4+ T cell lineage choice decisions. In mice, Itk mutations were shown to affect Th17-Treg lineage choice in favor of the latter. In this study, we explored whether human ITK reciprocally regulates Th17-Treg balance as its murine ortholog.Whole Exome Sequencing was used to identify the mutation. ITK-deficient peripheral blood lymphocytes were characterized by FACSAria III-based flow cytometric assays with respect to proliferation, apoptosis, cytokine production, and innate lymphoid cell (ILC) frequency. Sorted T cells from healthy donors were exposed to ibrutinib, an irreversible ITK inhibitor, to assess ITK's contribution to Th17 and Treg cell generation and functions.In this study, we report a child with a novel ITK mutation who showed impaired CD3/CD28 induced proliferation in T cells. ITK-mutant cells were more apoptotic irrespective of TCR activation. More importantly, T cells produced less Th17-associated cytokines IL-17A, IL-22, and GM-CSF. Conversely, Th1-associated IFN-γ production was increased. An irreversible inhibitor of ITK, ibrutinib, blocked ex vivo Th17 generation and IL-17A production, conversely augmented FOXP3 expression only at low doses in Treg cultures. Finally, we analyzed peripheral ILC populations and observed a relative decrease in ILC2 and ILC3 frequency in our ITK-deficient patient.To our knowledge, this is the first report showing that both genetic and chemical inhibition of ITK result in reduced Th17 generation and function in humans. We also report, for the first time, a reduction in ILC2 and ILC3 populations in an ITK-deficient human patient.