Ca2+− Axis−Controlled NFAT Nuclear Translocation Is Crucial for Optimal T Cell Immunity in an Early Vertebrate

This information is current as Xiumei Wei, Huiying Li, Yu Zhang, Cheng Li, Kang Li, of October 2, 2021. Kete Ai and Jialong Yang J Immunol published online 23 December 2019 http://www.jimmunol.org/content/early/2019/12/21/jimmun ol.1901065 Downloaded from

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The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2019 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published December 23, 2019, doi:10.4049/jimmunol.1901065 The Journal of Immunology

Ca2+–Calcineurin Axis–Controlled NFAT Nuclear Translocation Is Crucial for Optimal T Cell Immunity in an Early Vertebrate

Xiumei Wei,*,1 Huiying Li,*,1 Yu Zhang,* Cheng Li,* Kang Li,* Kete Ai,* and Jialong Yang*,†

Calcium ion (Ca2+) is a widespread and primitive second messenger that regulates physiological cell functions in almost all life beings. Ca2+ influx-induced NFAT activation is essential for T cell function and adaptive immunity. However, whether and how Ca2+ signaling modulates T cell immunity in early vertebrates, especially in nontetrapods, remains largely unknown. To address these questions, a Nile tilapia (Oreochromis niloticus) model was employed to investigate the regulation of ancestral T cell immunity by Ca2+–NFAT signaling in jawed fish. In Nile tilapia, an evolutionarily conserved Ca2+–NFAT signaling pathway is involved in the primary adaptive immune response during Streptococcus agalactiae infection. Meanwhile, T cell signals trigger Downloaded from several events along the Ca2+–NFAT axis in this early vertebrate, including Ca2+ influx, calcineurin activation, and NFAT nuclear import. More critically, suppression of Ca2+–NFAT signaling by the calcineurin inhibitor cyclosporine A impairs primordial T cell activation, clonal expansion, and infection clearance. Mechanistically, Nile tilapia NFAT interacts with several other transcription factors for potent expression, and T cells in this nontetrapod employ Cabin1 and DYRK1A to regulate NFAT nuclear import and export, respectively. To the best of our knowledge, this study is the first to demonstrate the regulatory mechanism of Ca2+–NFAT signaling on T cell immunity in a nontetrapod species. We suggest that modulation of T cell immunity by http://www.jimmunol.org/ Ca2+–NFAT signaling is a primitive strategy that already existed prior to the divergence of bony fish from the tetrapod lineage. The findings of this study provide valuable perspectives for understanding the evolution of adaptive immune system. The Journal of Immunology, 2020, 204: 000–000.

uring evolution, cells have adopted positively charged employ a number of components to assemble a Ca2+ signaling calcium ion (Ca2+) and negatively charged phosphate system with wide spatial and temporal dynamics (2). Among ions as two universal tools for signal transduction (1). these components, the Ca2+ release–activated calcium (CRAC) D 2+ by guest on October 2, 2021 Benefiting from the rapid switch between on and off statuses, channel, which mediates intracellular Ca increase, and its strong homeostasis via influx/efflux, and the ability to interact tightly regulated NFAT are of paramount importance for T cell with thousands of (2), Ca2+ acts as a versatile second immunity (3–5). messenger and plays pivotal roles in nearly every cellular process, Engagement of the TCR triggers the activation of including excitability, motility, metabolism, exocytosis, , tyrosine kinases, such as LCK and ZAP70, and eventually results transcription, and immunity (1, 2). To achieve this versatility, cells in tyrosine phosphorylation of Cg1 (PLCg1). Ac- tivated PLCg1 further hydrolyzes the membrane phospholipid phophatidylinositol-4,5-biphosphate (PIP2) and generates two sec- *State Key Laboratory of Estuarine and Coastal Research, School of Life Sciences, † ond messengers inositol 1,4,5-triphosphate (IP ) and diacylglycerol East China Normal University, Shanghai 200241, China; and Laboratory for Marine 3 Biology and Biotechnology, Qingdao National Laboratory for Marine Science and (DAG) (6, 7). DAG triggers the MAPK/ERK and PKCu-IKK-NF- Technology, Qingdao 266071, China kB pathways, whereas IP3 binds to the IP3 receptor (IP3R) on the 1X.W. and H.L. contributed equally to this work. endoplasmic reticulum (ER) membrane to permit Ca2+ efflux from 2+ 2+ ORCID: 0000-0002-3112-5012 (J.Y.). ER Ca stores into the cytoplasm (7). Depletion of ER Ca Received for publication August 30, 2019. Accepted for publication November 22, activates CRAC channel in the plasma membrane and induces 2019. sustained Ca2+ influx from extracellular compartment into the This work was supported by the National Natural Science Foundation of China cytoplasm, which is required for optimal TCR-induced signal (31972822 and 31872591), the Shanghai Pujiang Program (18PJ1402700), and the transduction (8, 9). An increase in intracellular Ca2+ is sensed by Fundamental Research Funds for the Central Universities. the EF hands of calcium-binding protein calmodulin, which in Address correspondence and reprint requests to Prof. Jialong Yang, East China Normal University, 500 Dongchuan Road, Shanghai 200241, China. E-mail address: turn activates activity of the serine/threonine phos- [email protected] phatase calcineurin. Activated calcineurin dephosphorylates cy- The online version of this article contains supplemental material. toplasmic NFAT, which exposes its nuclear localization sequence Abbreviations used in this article: Cabin1, calcineurin-binding protein 1; CRAC, Ca2+ and leads to rapid nuclear import (10). In the nuclear, NFAT co- release–activated calcium; DYRK1A, dual-specificity tyrosine phosphorylation-regulated operates with multiple transcription factors from other signaling kinase 1A; ECNU, East China Normal University; ER, endoplasmic reticulum; pathways, such as AP-1 and NF-kB, to induce transcription of GSK3, glycogen synthase kinase 3; IP3, inositol 1,4,5-triphosphate; IP3R, IP3 recep- tor; NCBI, National Center for Biotechnology Information; NJ, neighbor-joining; specific that control T cell activation, proliferation, and PLCg1, phospholipase Cg1; qPCR, quantitative real-time RT-PCR; RHD, Rel differentiation (10–13). homology domain; siRNA, small interfering RNA; Treg, regulatory T. NFAT-driven and T cell immunity are highly 2+ Copyright Ó 2019 by The American Association of Immunologists, Inc. 0022-1767/19/$37.50 dependent on sustained Ca influx and calcineurin activity. For

www.jimmunol.org/cgi/doi/10.4049/jimmunol.1901065 2Ca2+–NFAT IS CRUCIAL FOR EARLY VERTEBRATE T CELL IMMUNITY example, inhibition of CRAC channel impairs TCR