A novel mutation causing complete TYK2 deficiency, with severe respiratory viral infections, EBV-driven lymphoma, and Jamestown Canyon viral encephalitis

doi:10.21203/rs.3.rs-2817321/v1

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Research Article

A novel mutation causing complete TYK2 deficiency, with severe respiratory viral infections, EBV-driven lymphoma, and Jamestown Canyon viral encephalitis

https://doi.org/10.21203/rs.3.rs-2817321/v1

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Journal Publication

published 11 Sep, 2023

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Autosomal recessive Tyrosine kinase 2 (TYK2) deficiency is characterized by susceptibility to mycobacterial and viral infections. Here, we report a four-year-old female with severe respiratory viral infections, EBV-driven Burkitt-like lymphoma, and infection with the neurotropic Jamestown Canyon virus. A novel, homozygous c.745C > T (p.R249*) variant was found in TYK2. The deleterious effects of the TYK2 lesion were confirmed by immunoblot; by evaluating functional responses to IFN-α/β, IL-10, IL-12, and IL-23; and by assessing its scaffolding effect on cell surface expression of cytokine receptor subunits. The effects of the mutation could not be pharmacologically circumvented in vitro, suggesting that alternative modalities, such as hematopoietic stem cell transplantation or gene therapy, may be needed. We characterize the first patient from Canada with a novel homozygous mutation in TYK2.

Tyrosine kinase 2 (TYK2) is a member of the Janus kinase (JAK) family of kinases that, together with Signal Transducer and Activator of Transcription (STAT) molecules, constitute the JAK-STAT pathway, transducing signals downstream of multiple cytokine receptors to mediate innate and adaptive immune responses.

Rare biallelic, loss-of-function mutations in TYK2 define autosomal recessive TYK2 deficiency, an inborn error of immunity characterized by increased susceptibility to mycobacterial and/or viral diseases^1–3. The accompanying mycobacterial disease may be of the M. tuberculosis complex (e.g. M. tuberculosis sensu stricto, M. bovis Bacille Calmette-Guérin) or non-tuberculous mycobacteria (NTM). The viral diseases have included those that are DNA-based (e.g. Herpes simplex virus-1, Varicella-zoster virus, Molluscum contagiosum virus) as well as RNA-based (e.g. human Parainfluenza virus type 3, SARS-CoV-2, Influenza A). EBV-driven lymphoproliferative disorders have been observed⁴, as have atopic features (e.g. eczema, elevated serum IgE levels)¹. Loss of TYK2 function critically impairs type I interferon (IFN) responses (e.g. IFN-α, IFN-β), accounting for the predisposition to severe viral diseases, while poor IFN-γ responses, particularly to IL-23, underlie mycobacterial susceptibility.

Herein, we characterize a novel p.R249* mutation in TYK2 from a child with numerous life-threatening viral infections, while expanding the spectrum to include the neurotropic Jamestown Canyon virus.

Subjects

Subjects and the patient's family members provided informed consent on McGill University Health Centre (MUHC) institutional review board–approved research protocol (GEN10-256). Comprehensive medical histories, including review of all available outside records and serial clinical evaluations at the local institution, as well as immunologic laboratory testing at the MUHC were performed.

Sequencing and bioinformatics analysis

Whole exome sequencing (WES) was performed at a CLIA-certified diagnostic laboratory (GeneDx, Gaithersburg, Maryland, US). For Sanger sequencing, the TYK2 gene was PCR amplified from genomic DNA using primers designed to flank the respective regions (primers and sequencing conditions available on request). Sequencing was performed at the Génome Québec Innovation Centre. Sequencing analyses were performed on Sequencher® sequence analysis software (Gene Codes Corporation, Ann Arbor, MI).

Cell culture

EBV-induced lymphoblastoid cell lines (LCL) were established, as previously described^5-7. LCLs were cultured in RPMI supplemented with 10% fetal bovine serum (FBS), 20mM HEPES and penicillin/streptomycin (100U/ml and 100ug/ml respectively; Wisent). Immortalized human lung carcinoma epithelial cells, A549 and TYK2 knockout cell line A549 were constructed by CRISPR-Cas9 (Ubigene Biosciences) and cultured in a base medium F12/DMEM (Gibco/Invitrogen), containing 10% FBS. Cells were cultured at 37°C in a humidified CO₂ (5%) incubator. Cells were starved in serum-free RPMI and stimulated with IFNa/b (10⁵IU/ml), IL-10 (50ng/ml) or IL-12 (20ng/ml) for 30 min.

Antibodies and stimulation

Anti-TYK2, anti-phospho-Y1054/1055 TYK2, anti-myc tag, anti-phospho-p38 MAPK (Thr180/Thr182), anti-p38 were from Cell Signaling Technology. Anti-GAPDH was from Millipore. FITC-anti-phospho-STAT2 (Y699), APC-anti-phospho-STAT1 (Y701), PE-anti-phospho-STAT3 (Y705), APC-anti-phospho-STAT4 (Y693) and APC-anti-IFN-aR2 were from (Invitrogen). APC-anti-IL10RD, A488-anti-IL12Rb and A488-anti-IFNAR1 were from R&D system. Secondary antibodies for immunoblotting: DyLight800-anti-rabbit-IgG and DyLight680-anti-mouse-IgG were from ThermoScientific. Recombinant Human Interferon Alpha 2b and Human Interferon Beta 1b were purchased from PBL assay science. Recombinant human IL-12 and IL-10 were from PeproTech and recombinant human IL-23 from R&D system.

DNA constructs and transfection

The myc-flag tagged wild-type (WT) TYK2 plasmid was purchased from OriGene and confirmed WT by Sanger sequencing. The WT TYK2 plasmid was used to create the TYK2 c.745C>T variant using site-directed mutagenesis kit (New England Biolabs). Liposomal transfection in A549 cells was done with the Lipofectamine 3000 Transfection Reagent (Invitrogen) according to the manufacturer’s instructions. Cells were harvested 48 hours after transfection.

Immunoblotting

LCL or A549 (obtained by trypsinization) were resuspended in RIPA lysis buffer (Sigma) supplemented with cOmplete Mini Protease Inhibitor Cocktail (Roche) and PhosSTOP (Roche), then boiled in LDS Sample buffer and Blot Sample Reducing Agent (Thermo Fisher Scientific). Protein was separated by SDS-PAGE on premade gels (Novex) before transfer to membrane via iBlot Gel Transfer Device (Invitrogen). Membrane was blocked in 5% skim milk TBST to block nonspecific protein binding. The membrane was blotted using primary antibodies overnight at 4°C. The membrane was washed 4 times with 0.1% Tween 20 in TBS and then incubated with a 1:15,000 dilution of secondary antibodies in blocking buffer. Membranes were scanned and analyzed with an Odyssey IR scanner using Odyssey imaging software 3.0 (LI-COR Biosciences, Inc).

Quantitative real-time PCR

RNA was extracted using Trizol Reagent (Invitrogen) according to manufacturer’s instructions and quantified by NanoDrop-100 spectrophotometer. 500ng of RNA was reverse transcribed using the Maxima H Minus First Strand cDNA Synthesis Kit (Thermo Fisher Scientific). Real-time quantitative PCR was performed with CFX Opus 96 Real-Time PCR (Bio-Rad Laboratories). TaqMan quantitative PCR assays were run using primers/probes: CXCL11 (Hs00171138), IRF1 (Hs00971965), MX1 (Hs00895608), OAS2 (Hs00942643), ISG15 (Hs00192713), IL-8 (Hs00174103) and GAPDH (Hs02758991). The mRNA input was normalized to the expression of the housekeeping genes, GAPDH⁸. Statistical significance of the data was validated using One-way ANOVA with Tukey’s post hoc test and multiple comparison procedure. The level of significance was set at *p < 0.05.

Flow Cytometry

Cells were blocked in Fc Receptor Binding Inhibitor Monoclonal Antibody (Invitrogen) and incubated with LIVE/DEAD Fixable Dead Cell Stain (Thermo Fisher Scientific). For intracellular staining, cells were stained using the FIX & PERM Cell Fixation and Cell Permeabilization Kit (Thermo Fisher Scientific). Samples were analyzed using 4-laser Cytek Aurora flow cytometer (Cytek Biosciences). Data was done using FlowJo (Ashland, OR). For statistical analysis, repeated-measures one-way ANOVA, followed by Tukey's multiple comparison post-test was used. The level of significance was set at *p < 0.05.

Case Report

Patient 1 (P1) is a 4-year-old female born to parents of Pakistani origin with distant consanguinity (see Supplemental Appendix for complete clinical details). At 6 weeks of age, she developed respiratory failure due to influenza B, from which she recovered. Over the next few months, she developed recurrent respiratory infections requiring supplemental oxygen and treatment with bronchodilators and systemic corticosteroids. At 8 months of age, she was diagnosed with bilateral subdural hygromas, which eventually resolved. At 11 months old, she developed severe respiratory disease due to Enterovirus/Rhinovirus requiring hospitalization and oxygen supplementation, but not mechanical ventilation, from which she recovered. She received all routine immunizations, including live viral vaccines, without adverse events; she never received BCG vaccination. At age 4 years, she developed a rapidly-growing neck mass that invaded the left sternocleidomastoid, mediastinum and retroperitoneum. A mass was also noted in the recto-uterine pouch. Analysis of the mass revealed Burkitt lymphoma, with tumour cells positive for EBER; bacterial, mycobacterial, and fungal cultures yielded no organism. She was treated with R-COPADM-1 protocol (rituximab; vincristine; prednisone; methotrexate; folinic acid; cyclophosphamide; doxorubicin; cytarabine) followed by R-CYM2 (rituximab; methotrexate; folinic acid; cytarabine). During her active chemotherapy, she developed multiple viral infections including Coronavirus OC43/HKU1, rotavirus and norovirus gastroenteritis and enterovirus/rhinovirus URTI. In the context of screening for high-risk exposure, she was found to be PCR-positive for SARS-CoV-2 (B.1.1.529 VOC; Omicron); she manifested only with coryza. Five weeks later, her PCR test reverted to negative. Two weeks later, though, she had new cough and fever, and was SARS-CoV-2 PCR positive again; she did not require oxygen supplementation and resolved without complications. Three months after successful completion of her chemotherapy, she developed a febrile, tonic-clonic seizure. Imaging revealed focal areas of T2/T2 FLAIR hyperintensity and edema, with corresponding restricted diffusion in the cortex of bilateral parieto-occipital and posterior temporal lobes. Cerebrospinal fluid PCR analysis yielded Jamestown Canyon virus; Jamestown Canyon serology was IgM Positive by ELISA and 1:40 by plaque reduction neutralization test. She clinically stabilized but was left with significant neurologic impairment. Given her history and fulminant presentation, further investigations were performed (Immunophenotyping data, Table 1).

Table 1: Laboratory findings

Parameter	Reference range	June 18 2022***	August 20 2022	February 22 2023
WBC	4.7 - 13.5 10ˆ9/L	7.9	7.6	17.0 (H)
Hemoglobin	105 - 135 g/L	114	129	138 (H)
Platelets	150 - 450 10ˆ9/L	367	410	333
Neutrophils	1.5 - 8.5 10ˆ9/L	5.8	4.9	11.1 (H)
Lymphocytes	1.0 - 5.5 10ˆ9/L	1.2	1.6	3.2
Monocytes	0.1 - 1.0 10ˆ9/L	0.9	0.5	1.2 (H)
Eosinophils	0.0 - 0.6 10ˆ9/L	0.0	0.5	1.3 (H)
Basophils	0.0 - 0.1 10ˆ9/L	0.1	0.1	0.1
IgG	5.3 - 12.8 g/L	3.3 (L)	9.5*	10.1*
IgA	0.20 - 1.50 g/L	0.33	0.34	0.20
IgM	0.50 - 2.00 g/L	0.25 (L)	0.90	0.60
IgE	<=144.0 ug/L	336.0 (H)		101.0
% Total CD3	43.0 - 76.0 %	82.8 (H)	74.2	68.0
# Total CD3	900-4,500 10ˆ6/L	1,385	1,391	1816
CD3/CD4%	23.0 - 48.0 %	20.5 (L)	25.1	22.1 (L)
CD3/CD4	700-2,200 10ˆ6/L	343 (L)	470 (L)	589
CD3/CD8%	14.0 - 33.0 %	55.7 (H)	40.7 (H)	1054
CD3/CD8	300-1,600 10ˆ6/L	931	764	0.56 (L)
CD4:CD8 Ratio	0.9 - 2.9	0.4 (L)	0.6 (L)	21.4
% Total CD19	14.0 - 44.0 %	2.9 (L)**	10.2 (L)	573.0
# Total CD19	200.0-2,100.0 10ˆ6/L	45.8 (L)**	189.8 (L)	10.8
% Total CD(16+56) Pos CD3 Neg	4.0 - 23.0 %	12.9	16.8	287
# Total CD(16+56) Pos CD3 Neg	100-1,000 10ˆ6/L	202	313	68.0

* Post IVIG

** Had received a dose of rituximab 3 months prior

*** Post high dose corticosteroids during acute treatment of encephalitis

(H): Higher than the upper limit of the reference range

(L): Lower than the lower limit of the reference range

Homozygous p.R249* variant in TYK2

Whole exome sequencing (WES) in P1 identified homozygous c.745C>T (p.R249*) in TYK2 (NM_003331). This variant, which has not been previously reported, was confirmed by Sanger sequencing [Figure 1A]. It is predicted to cause a premature termination codon in exon 7 residing in the FERM domain (aa 28-451), which, together with the adjacent SH2-like domain, mediate its interaction with receptors [Figure 1B]; in particular, this region of TYK2 is necessary for signaling through the type I IFN receptor system⁹. This variant was predicted in silico to be deleterious (e.g. SIFT, PolyPhen2; CADD Phred; MutationTaster) and is extremely rare (gnomAD allele frequency of 4.13x10^-6; no homozygotes reported).

To confirm the predicted pathogenicity of the variant, we first assessed its molecular impact on TYK2 expression by immunoblot of EBV-transformed B-lymphoblastoid cell lines (LCLs) from P1 vs. health controls [HC], showing a loss of expression in P1 [Figure 1C]. We confirm this result in an autonomous A549 cell model system in which endogenous TYK2 was knocked out by CRISPR-Cas9-based technology, then transiently transfected with either wild-type (WT) TYK2 allele or the c.745C>T variant (TYK2 p.R249*) [Figure 1D]. Both immunoblotting against the TYK2 protein or against the C-terminal-myc-tag showed no TYK2 protein detected from the c.745C>T allele, confirming its loss of expression.

Kinase Activity of TYK2 p.R249*is impaired in P1’s LCL

We first analysed the capacity for auto- and trans-phosphorylation of TYK2 by immunoblotting in P1 and HC LCLs following stimulation with either IFNa/b, IL-10 or IL-12. In contrast to responses seen in HC, the absence of TYK2 expression, as well as its phosphorylation, were observed in P1's LCLs [Figure 2A].

We then assessed the impact of TYK2^R249* on its downstream kinase activity within the JAK/STAT canonical signaling pathway, by flow cytometry-based measurement of phosphorylation of the respective STAT proteins that mediate signaling from IFNa/b, IL-12, IL-23 and IL-10 [Figure 2B]. For all tested stimuli, P1 LCLs show reduced STAT activation relative to controls, consistent with its lack of TYK2 protein expression. To confirm these anomalies in a patient-independent cell model, an A549 cell line with TYK2 knocked out was then complemented with either empty vector (EV), TYK2 WT or p.R249* construct through transient transfection. No increase in phosphorylated STAT1 or STAT2 via IFNa/b stimulation or of STAT4 in response to IL12 was observed in A549 cells with either EV or bearing the p.R249* allele.

We further evaluated the p38-mediated non-canonical signaling pathway for type I IFN by assessing phosphorylation of p38 in A549 cell lines, given the high basal phospho-p38 (p-p38) activity in LCL. In the absence of TYK2 or in the presence of TYK2^R249*, the phosphorylation of p38 is very low in response to IFN stimulation [Figure 2D].

These results indicate that the identified c.745C>T variant in TYK2 results in complete loss of all its known kinase activity in various human cytokine signaling pathways.

Scaffolding function of TYK2 on the expression of associated cytokine receptors

The FERM and SH2 domains of TYK2 also possess scaffolding function^{2, 10, 11}, and previous patients with TYK2 deficiency have decreased cell surface expression of the cytokine receptor subunits, IFN-αR1, IFN-αR2, IL-10R2, and IL-12Rβ1³. The absence of the SH2 domain and part of the FERM domain in the p.R249* mutant may similarly abolish TYK2's interaction with these molecules, thus decreasing their stability and cell surface expression. To evaluate this, we measured the surface expression and intracellular levels of IFN-αR1, IFN-αR2, IL-10R2, and IL-12Rβ1 on LCL by flow cytometry [Figure 3]. As previously shown, cell surface expression of IFN-αR1, IFN-αR2, IL-10R2, and IL-12Rβ1 receptor subunits were significantly reduced in P1’s LCL relative to controls [Figure 3, left column], as were their intracellular levels [Figure 3, right column], confirming that the p.R249* allele causes loss of TYK2's scaffolding function. In contrast, cell surface and intracellular levels of IFN-αR2 were comparable between P1 and controls.

Impact of TYK2-deficient patient P1 on ISGs expression

Type I interferon stimulation leads to the expression of IFN-stimulated genes (ISGs), which may be functionally categorized as "robust" (primarily exerting antiviral effects) and "tunable" (primarily mediating proinflammatory or antiproliferative effects)^{12, 13}. Given the severely diminished activation of TYK2/STAT1/STAT2 following IFNa/b-stimulation of P1's LCLs, we analyzed the impact of TYK2^R249* on expression of select ISG by RT-qPCR of "robust" (i.e. OAS2, ISG15, MX1) and "tunable" (i.e. IL-8, CXCL11, IRF1) genes [Figure 4A]. In this LCL model, we observed a clear decrease in the expression of the antiviral and antiproliferative ISGs, except for IRF1. We confirmed this observation in A549 TYK2^-/- cells complemented with WT or mutant TYK2 [Figure 4B]. Thus, the c.745C>T mutation in TYK2 impairs ISG responses.

Attempts for pharmacological correction in vitro

Given the dire clinical susceptibility to common viruses in this young child, we investigated whether the mutant TYK2 could be pharmacologically circumvented. G418, an aminoglycoside, has previously been shown to mediate ribosomal in-frame read-through of nonsense mutations^14-16, while 4-phenylbutyrate (4PBA) is a chemical chaperone that can rescue the expression of mutant proteins normally expressed at the plasma membrane by improving their intracellular trafficking^17-19. Neither agent alone, or in combination, restored TYK2 expression in P1's LCL (Supplementary figure S1). Since IFN-g stimulation of its cognate receptor can activate JAK1 and STAT1, which are shared with the IFN-a/b signaling pathway, we sought to determine if IFN-g may offset the diminished ISGs caused by TYK2^R249*. Of the genes tested, IFN-g stimulation compensated some of the tunable ISGs (IRF1, IL-8), but none of the robust, antiviral ISGs (Supplementary figure S2).

Different sub-types of TYK2 deficiency have been recently defined, based on molecular characteristics^2-4: Complete deficiency with no TYK2 protein production (loss of expression, LOE, and loss of function, LOF); complete deficiency with detectable protein (residual expression but LOF); partial deficiency due to hypomorphic mutation; partial TYK2 deficiency specifically compromising IL-23 responses due to the common p.P1104A variant; and partial TYK2 deficiency specifically compromising IL-23 responses due to rare variants. Here, we present a case with a novel homozygous c.745C>T (p.R249*) mutation, leading to loss of expression and consequently, loss of kinase and scaffolding functions, resulting in complete TYK2 deficiency (LOE, LOF).

Numerous respiratory and gastrointestinal viral infections occurred in P1, including multiple episodes of life-threatening disease, in keeping with previous reports. Intriguingly, infection with SARS-CoV-2 only resulted in non-severe/critical disease, despite not having been vaccinated against COVID-19 nor having received monoclonal or polyclonal immunoglobulin infusions. Six other pediatric patients with TYK2 deficiency who contracted SARS-CoV-2 infection have been previously reported²⁰; all had complete deficiency (LOE, LOF) due to either c.647delC (p.P216Rfs*14) in 5 or homozygous c.466-1G>A in 1; four had hypoxemic COVID-19 pneumonia. The remaining two patients, along with this one reported here, may suggest that life-threatening COVID-19 shows incomplete penetrance in those with TYK2 deficiency. On the other hand, the patient reported here developed a severe meningoencephalitis with Jamestown Canyon virus (JCV), an arthropod-borne RNA-based virus (arbovirus) belonging to the California serogroup of the Bunyaviridae family within the genus Orthobunyavirus²¹. JCV is maintained in an enzootic transmission cycle involving primary reservoir and amplifying hosts (e.g. white-tailed deer) and mosquito vectors. Seroprevalence studies suggest that many, perhaps as much as 50% of JCV infections, are a-/pauci-symptomatic^{22, 23}. Whether P1's severe JCV central nervous system (CNS) disease was due to her underlying TYK2 deficiency, previous chemotherapy, and/or other factors is unknown at this time. However, it is notable that other CNS infections have been reported in patients with TYK2, due to Herpes virus or Brucella², suggesting that TYK2 may underlie additional cases of severe CNS infections; further reports will help resolve this issue.

Given the centrality of TYK2 to signaling downstream from multiple receptors, we demonstrate impaired stimulation of the type I IFN receptor (IFN-I-R), IL-10R, IL-12R, and IL-23R, similarly to what has been previously shown^{1-3, 24}. In line with previous reports of TYK2 deficient patients, and with other inborn errors of IFN-I immunity^{25, 26}, the loss of IFN-I-R accounts for P1's unusual susceptibility to severe viral disease. Further support for this is the loss of "robust" interferon stimulated genes (ISGs), represented by expression of OAS2, ISG15 and MX1, which are primarily antiviral in action^{12, 13}. On the other hand, "tunable" ISGs are considered to primarily arbitrate immunomodulatory and antiproliferative functions^{12, 13}, and impaired expression of these genes may underlie the EBV-positive Burkitt lymphoma in P1 and EBV-driven lymphoproliferative disorder in other TYK2 deficiency patients⁴. Interestingly, EBV encodes latent membrane protein 1 (LMP-1), an oncoprotein essential for B cell immortalization, and LMP-1 targets TYK2²⁷. Although loss of IL-23 function predisposes to mycobacterial disease²⁸, particularly of the M. tuberculosis complex, P1 has not manifested such disease because she did not receive BCG vaccination (in keeping with provincial public health policy), and tuberculosis is not endemic where she lives. Similarly, impaired IL-12R function increases susceptibility to disseminated salmonellosis or thermally-dimorphic endemic mycoses^29-33, neither of which are prevalent in P1's region of residence. Thus, P1 serves as a reminder that the infectious disease phenotype requires specific microbial exposures on a permissive immunologic background.

TYK2's scaffolding function is required for the stable surface expression of IFNAR1, IL10R2, and IL12RB1^{3, 34-38}. We observed diminished cell surface expression of these proteins, along with decreased intracellular levels, likely due to their instability from loss of TYK2 and their subsequent degradation. On the other hand, IFNAR2 is present in comparable quantities at the cell surface between P1 and controls, although it is insufficient to mediate signaling. Attempts to restore TYK2, with translational read-through of the premature stop codon and chaperone treatment with 4PBA, were unsuccessful, and did not improve type I IFN signaling (data not shown). Similarly, treatment with IFN-g only had modest correction of select ISG. Collectively, these findings would argue that pharmacologic doses of IFN may be futile clinically and that genetic-based therapy or hematopoietic stem cell transplantation may be urgently required to correct the immunodeficiency of complete TYK2 deficiency.

Acknowledgement

We thank the patient and their family for participating in our research. We thank the interdisciplinary medical personnel involved in the care of the patient and his family.

Funding

This work was supported by the McGill University Health Centre – Foundation via the SDR Project.

D.C. Vinh is supported by the Fonds de Recherche du Québec - Santé (FRQS) – Senior clinician-scientist scholar program.

Conflicts of interest/Competing interests

D.C. Vinh has received research support, unrestricted educational grants, consultancy fees, and speaker honoraria from CSL Behring; has received consultancy fees from AstraZeneca, Moderna, and Takeda; and has undertaken clinical trials for Cidara Therapeutics and Moderna. The remaining authors declare no competing financial interests.

Availability of data and material

All data generated or analyzed during this study are included in this published article and its supplementary information files.

Authors' contributions

Study design and supervision were performed by Lucie Roussel, Anne Pham-Huy, and Donald C. Vinh. Clinical data generation, acquisition, and analysis were performed by Anne Pham-Huy, Andrea C. Yu, Sunita Venkateswaran, and Justin Penner. Experimental data was done by Lucie Roussel, Anna Perez, Guillaume Bourdel, Yichun Sun, Stephanya Tellez Villavicencio, Stéphane Bernier, Yongbiao Li, Makayla Kazimerczak-Brunet, Rolan Alattar, Marc-André Déry, and Adam J. Shapiro. Lucie Roussel, Anne Pham-Huy, and Donald C. Vinh wrote the first draft of the manuscript. All authors have reviewed, contributed to and approved the final version of the manuscript. Donald C. Vinh is the principal investigator providing supervision and funding.

Ethics approval

The Research Ethics Board of the Research Institute – McGill University Health Centre has approved this research (No. 2021-6624)

Consent to participate

The authors confirm that a relevant local consent has been signed by the patient and/or the legally authorized representative.

Consent for publication

The authors confirm that a relevant local consent has been signed by the patient and/or the legally authorized representative.

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published 11 Sep, 2023

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Editorial decision: Major Revisions Needed
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A novel mutation causing complete TYK2 deficiency, with severe respiratory viral infections, EBV-driven lymphoma, and Jamestown Canyon viral encephalitis

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Abstract

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Introduction

Materials And Methods

Cell culture

Immunoblotting

Quantitative real-time PCR

Results

Kinase Activity of TYK2 p.R249*is impaired in P1’s LCL

Discussion

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References

Supplementary Files

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Journal Publication

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