Innate type 2 lymphocytes (ILC2, also called natural helper cells or nuocytes) are critically involved in widespread infectious and allergic diseases. By residing at mucosal barriers and responding to epithelial-derived cytokines such as IL-33 and IL-25, they rapidly release the early type 2 cytokines that promote the Th2 inflammatory responses of adaptive CD4 T cells and the IgE production of B cells. However, the definition of the ILC2 lineage remains tentative and somewhat controversial in the absence of molecular signature. Furthermore, while they derive from the same common lymphoid precursors (CLP) as other adaptive and innate lymphocytes, their developmental intermediates and their relationship with other related lineages are unknown. This project builds on preliminary studies of PLZF-IRES-GFPCre reporter mice produced in our laboratory showing that the transcription factor PLZF, previously identified as the signature of the innate-like IL- 4/IL-13-producing NKT cell lineage, is also expressed at high levels during the development of ILC2s. The central hypothesis is that PLZF exerts critical functions in the development of ILC2s and that PLZF- reporter/deleter mice can be used to genetically track and manipulate ILC2 precursors and their mature progeny. The objective of this application is to use PLZF-IRES-GFPCre mice to identify the bone marrow precursors of ILC2, characterize their molecular program and define their progeny through fate mapping after crossing to ROSA26-FL-STOP-FL-Tomato mice. Furthermore, the project will test various approaches to genetically manipulate or ablate ILC2 in vivo as a prelude to characterizing their function in physiology and in allergic airway inflammation.
The specific aims are: 1) to characterize the PLZFhi subset of CLP;2) to determine the role of PLZF in ILC2 development and function;3) to create ILC2-defective mouse models for studies of allergic airway inflammation. The project is innovative because it explores the novel idea that PLZF expression is shared across innate and innate-like lineages specializing in type 2 responses and because it will produce new models and reagents for studies of ILC2s in vivo. This would signal a major shift in our ability to understand and further manipulate the role of innate immunity in worldwide diseases caused or cured by type 2 immunity. The proposed research is significant because it will considerably enhance our understanding of innate lymphocyte lineage development. The concepts and tools developed will make it possible to develop agents that target distinct components of innate immunity to parasites and allergens.

Public Health Relevance

The proposed research will generate new tools and concepts to understand the development and function of a recently discovered population of immune cells called innate type 2 lymphocytes (ILC2) that is critically involved in allergic diseases such as asthma as well as in the immune rejection of helminthic parasites. It is relevant to public health because the results will allow the development of new approaches to prevent or correct defects leading to common allergic diseases or to the control of parasitic diseases. Thus, this work will directly support the overall NIH mission of developing fundamental knowledge that will help reduce the burden of human disease and, specifically, prevent and treat allergic pulmonary diseases.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Project (R01)
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Special Emphasis Panel (ZRG1-CVRS-H (02))
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Noel, Patricia
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University of Chicago
Schools of Medicine
United States
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Verhoef, Philip A; Constantinides, Michael G; McDonald, Benjamin D et al. (2016) Intrinsic functional defects of type 2 innate lymphoid cells impair innate allergic inflammation in promyelocytic leukemia zinc finger (PLZF)-deficient mice. J Allergy Clin Immunol 137:591-600.e1
Ishizuka, Isabel E; Constantinides, Michael G; Gudjonson, Herman et al. (2016) The Innate Lymphoid Cell Precursor. Annu Rev Immunol 34:299-316
Ishizuka, Isabel E; Chea, Sylvestre; Gudjonson, Herman et al. (2016) Single-cell analysis defines the divergence between the innate lymphoid cell lineage and lymphoid tissue-inducer cell lineage. Nat Immunol 17:269-76
Constantinides, Michael G; Gudjonson, Herman; McDonald, Benjamin D et al. (2015) PLZF expression maps the early stages of ILC1 lineage development. Proc Natl Acad Sci U S A 112:5123-8
Bunker, Jeffrey J; Flynn, Theodore M; Koval, Jason C et al. (2015) Innate and Adaptive Humoral Responses Coat Distinct Commensal Bacteria with Immunoglobulin A. Immunity 43:541-53
Constantinides, Michael G; McDonald, Benjamin D; Verhoef, Philip A et al. (2014) A committed precursor to innate lymphoid cells. Nature 508:397-401