Inborn errors in RNA polymerase III underlie severe varicella zoster virus infections
Benson Ogunjimi, … , Søren R. Paludan, Trine H. Mogensen
Benson Ogunjimi, … , Søren R. Paludan, Trine H. Mogensen
Published August 7, 2017
Citation Information: J Clin Invest. 2017;127(9):3543-3556. https://doi.org/10.1172/JCI92280.
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Research Article Genetics Infectious disease

Inborn errors in RNA polymerase III underlie severe varicella zoster virus infections

Abstract

Varicella zoster virus (VZV) typically causes chickenpox upon primary infection. In rare cases, VZV can give rise to life-threatening disease in otherwise healthy people, but the immunological basis for this remains unexplained. We report 4 cases of acute severe VZV infection affecting the central nervous system or the lungs in unrelated, otherwise healthy children who are heterozygous for rare missense mutations in POLR3A (one patient), POLR3C (one patient), or both (two patients). POLR3A and POLR3C encode subunits of RNA polymerase III. Leukocytes from all 4 patients tested exhibited poor IFN induction in response to synthetic or VZV-derived DNA. Moreover, leukocytes from 3 of the patients displayed defective IFN production upon VZV infection and reduced control of VZV replication. These phenotypes were rescued by transduction with relevant WT alleles. This work demonstrates that monogenic or digenic POLR3A and POLR3C deficiencies confer increased susceptibility to severe VZV disease in otherwise healthy children, providing evidence for an essential role of a DNA sensor in human immunity.

Authors

Benson Ogunjimi, Shen-Ying Zhang, Katrine B. Sørensen, Kristian A. Skipper, Madalina Carter-Timofte, Gaspard Kerner, Stefanie Luecke, Thaneas Prabakaran, Yujia Cai, Josephina Meester, Esther Bartholomeus, Nikhita Ajit Bolar, Geert Vandeweyer, Charlotte Claes, Yasmine Sillis, Lazaro Lorenzo, Raffaele A. Fiorenza, Soraya Boucherit, Charlotte Dielman, Steven Heynderickx, George Elias, Andrea Kurotova, Ann Vander Auwera, Lieve Verstraete, Lieven Lagae, Helene Verhelst, Anna Jansen, Jose Ramet, Arvid Suls, Evelien Smits, Berten Ceulemans, Lut Van Laer, Genevieve Plat Wilson, Jonas Kreth, Capucine Picard, Horst Von Bernuth, Joël Fluss, Stephane Chabrier, Laurent Abel, Geert Mortier, Sebastien Fribourg, Jacob Giehm Mikkelsen, Jean-Laurent Casanova, Søren R. Paludan, Trine H. Mogensen

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Figure 4

Impaired production of immunostimulatory RNA by patient cells after transfection with AT-rich DNA.

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Impaired production of immunostimulatory RNA by patient cells after tran...
(A) Schematic illustration of the pathways through which poly(dA:dT) and poly(I:C) induce IFN expression. (B) Poly(dA:dT) beads incubated with cytoplasmic extracts of PBMCs from healthy controls, P1, P2, and P3 were precipitated (precip) and immunoblotted with anti-POLR3A and anti-POLR3C. (C) Overview of the assay used to evaluate for POL III activity. (D and E) PBMCs from P1, P2, P3 and 4 healthy controls were transfected with poly(dA:dT) or VZV-AT-high DNA (2 μg/ml). Total RNA was isolated 14 hours later and transfected into HEK293 cells. Total RNA was isolated 6 hours later, and levels of IFNB mRNA were determined by RT-qPCR. The levels of IFNB mRNA were normalized to GAPDH, and data are presented as mean ± SD from 3 replicates from 2 independent experiments. Lipo, Lipofectamine. *0.01 < P < 0.05; **0.01 < P < 0.001; ***0.001 < P < 0.0001. Groups were compared pairwise using post hoc t tests based on data from all groups and 1-way ANOVA.