Abstract
Herpes simplex virus type 1 (HSV-1) encephalitis (HSE) is the most common sporadic viral encephalitis in Western countries. Some HSE children carry inborn errors of the Toll-like receptor 3 (TLR3)-dependent IFN-α/β– and -λ–inducing pathway. Induced pluripotent stem cell (iPSC)-derived cortical neurons with TLR3 pathway mutations are highly susceptible to HSV-1, due to impairment of cell-intrinsic TLR3-IFN immunity. In contrast, the contribution of cell-intrinsic immunity of human trigeminal ganglion (TG) neurons remains unclear. Here, we describe efficient in vitro derivation and purification of TG neurons from human iPSCs via a cranial placode intermediate. The resulting TG neurons are of sensory identity and exhibit robust responses to heat (capsaicin), cold (icilin), and inflammatory pain (ATP). Unlike control cortical neurons, both control and TLR3-deficient TG neurons were highly susceptible to HSV-1. However, pretreatment of control TG neurons with poly(I:C) induced the cells into an anti–HSV-1 state. Moreover, both control and TLR3-deficient TG neurons developed resistance to HSV-1 following pretreatment with IFN-β but not IFN-λ. These data indicate that TG neurons are vulnerable to HSV-1 because they require preemptive stimulation of the TLR3 or IFN-α/β receptors to induce antiviral immunity, whereas cortical neurons possess a TLR3-dependent constitutive resistance that is sufficient to block incoming HSV-1 in the absence of prior antiviral signals. The lack of constitutive resistance in TG neurons in vitro is consistent with their exploitation as a latent virus reservoir in vivo. Our results incriminate deficiencies in the constitutive TLR3-dependent response of cortical neurons in the pathogenesis of HSE.
Original language | English (US) |
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Pages (from-to) | E8775-E8782 |
Journal | Proceedings of the National Academy of Sciences of the United States of America |
Volume | 115 |
Issue number | 37 |
DOIs | |
State | Published - Sep 11 2018 |
Funding
ACKNOWLEDGMENTS. We thank the patients and their families for participating in this study; Dr. Nai-Kong Cheung (Memorial Sloan Kettering Cancer Center) for the 3F8 (clinical grade GD2) antibody; Sanghoon Oh (Memorial Sloan Kettering Cancer Center Molecular Cytology Core) for excellent technical support in calcium imaging experiments and writing of the Matlab code; and Sarah Antinone (Northwestern University) for help with HSV-1 data analysis. The work was funded by NIH Grant R01NS072381. Additional support was provided by NIH Grant R21NS084255, NIH Cancer Center support Grant P30CA008748, Clinical and Translational Science Award program Grant UL1TR001866, Agence Natio-nale de Recherches Grant IEIHSEER, the Rockefeller University, and INSERM. B.Z. was supported by a NYSTEM postdoctoral fellowship. O.E. received support from the NIH-National Institute of Allergy and Infectious Diseases Immunology and Molecular Pathogenesis Training Program and The American Society for Microbiology Robert Watkins fellowship. L.D.N. is supported by the Division of Intramural Research, National Institute of Allergy and Infectious Diseases, NIH.
Keywords
- Antiviral immunity
- HSE
- HSV-1
- TG neurons
- TLR3
ASJC Scopus subject areas
- General