Le virus Herpes Simplex de type 1 (HSV-1) possède un mode d’infection particulier dit bimodal. Il peut soit se répliquer de manière active lors d’une phase dite lytique soit migrer dans les neurones et rester en latence. Il peut réactiver pour rétablir une infection lytique. Une protéine virale majeure dans la réactivation de HSV-1 est ICP0. C’est une protéine nucléaire à activité E3 ubiquitine ligase, qui possède la particularité d’induire la dégradation par le protéasome de plusieurs protéines centromériques constitutives, ce qui provoque une déstabilisation du centromère interphasique. Mon équipe a découvert une réponse cellulaire à l’instabilité centromérique, induite par la protéine ICP0, et nommée iCDR (pour interphase Centromere Damage Response.). L’objectif général de ma thèse est de déterminer les modifications structurales que subissent les centromères endommagés par ICP0 à l’origine de l’iCDR et probablement de la réactivation. J’ai pu démontrer qu’ICP0 affectait toute la structure protéique étroitement associée aux centromères durant l’interphase. Suite à ces résultats, j’ai pu démontrer, par des analyses de digestion de chromatine à la nucléase microccocale (MNAse), que l’occupation nucléosomique de la chromatine centromérique suite à l’activité d’ICP0 était affectée de façon significative. Une étude in vivo effectuée à partir de tissus nerveux provenant de souris infectées de manière latente, a permis de démontrer une co-localisation entre les génomes HSV-1 latents et les centromères. Cette co-localisation est associée à une répression transcriptionnelle du virus. Les résultats de ma thèse montrent donc que les effets d’ICP0 sur la déstabilisation des centromères sont en relation avec un rôle de ces centromères durant la latence. Ceci suggère fortement une implication de la déstabilisation des centromères dans le processus de réactivation contrôlé par ICP0. / The Herpes Simplex type 1 (HSV-1) virus possesses a bimodal mode of infection. It can either replicates in an active way during the lytic cycle, or it can infect neurons and stay in latency. HSV-1 reactivates from latently infected neurons for re-establishing a lytic infection. A major viral protein implicated in reactivation is ICP0. It is a nuclear E3 ubiquitin-ligase, which has the particularity to induce the proteasome-mediated degradation of several constitutive centromeric proteins. This activity severely destabilizes the interphase centromere. My team has discovered a novel cellular response triggered by the estabilization of centromeres by ICP0, called iCDR (interphase Centromere Damage Response). The general aim of my thesis is to determine the centromere structural modifications induced by ICP0 that can trigger the iCDR and probably the reactivation. I was able to demonstrate that ICP0 affected the entire proteinacious structure of interphase centromeres. Following this, I showed by micrococcal nuclease (MNase) digestion approach that the nucleosomal organization of centromeric chromatin was significantly affected by ICP0. An in vivo study in nervous tissues coming from latently infected mice enabled to show a co-localization between latent HSV-1 genomes and centromeres. This co-localisation is linked to a transcriptional repression of the virus. The results of my thesis show that the destabilization of centromere by ICP0 correlates with a role of the centromeres during latency. This strongly suggests an implication of centromere destabilization in the ICP0-controlled reactivation process.

A dor é descrita como uma sensação sensorial e emocional desagradável de extrema importância para sobrevivência e integridade do organismo. As dores crônicas de origem neuropática são de difícil tratamento e seus mecanismos fisiopatológicos pouco conhecidos. Este estudo foi realizado após inoculação do vírus HSV-1 na pata traseira esquerda de camundongos machos da linhagem Balb/C, e teve como objetivo investigar comportamentalmente o desenvolvimento de alodínia mecânica e hipernocicepção nas fases herpética e pós herpética, caracterizar a atividade de vias intracelulares de sinalização das proteínas JNK, AKT CREB, P38, ERK, Glutamina Sintetase e Stat 3, através de western blot na coluna dorsal da medula espinal nas fases herpética e pós herpética, além de verificar a presença do vírus HSV-1 na medula espinal e gânglio dos animais por meio da reação em cadeia da polimerase em tempo real (RT-PCR). Os resultados evidenciaram alteração de sensibilidade nos animais a partir do 7º dia que permaneceu até o 28º dia após a inoculação do vírus. Houve uma mudança na expressão das proteínas MAPKs, com aumento na expressão de JNK, AKT e CREB no corno dorsal da medula no 8º e 21º dia após a inoculação do vírus HSV-1, aumento na expressão de P 38 e P ERK no 8º dia após inoculação do vírus e uma diminuição na expressão da proteína Stat 3 no 8º e 21º dia após a inoculação do vírus, sugerindo assim a participação dessas proteínas na alteração de sensibilidade tanto no período herpético quanto pós herpético. Também ocorreu um aumento na amplificação do DNA viral HSV-1 na medula espinal e gânglio espinal esquerdo no período herpético após inoculação do vírus HSV-1 / The pain is described as a sensorial and emotional unpleasant sensation of extreme importance for survival and integrity of the organism. The chronic pains that has neuropathic source are hard to treat and its physiopathologic mechanisms not well known. This study was performed after inoculation of the virus HSV-1 in the left back foot of male Balb/C mouse, and had as main objectives to behaviorally investigate the development of mechanical allodynia and hyper nociception during the herpetic and post-herpetic phase, characterize the activity of the intracellular signalization paths of the proteins JNK, AKT CRB, P38, ERK, glutamine synthetase and Sat 3 during herpetic and post-herpetic phase using Western Blot, besides checking as well for the presence of HSV-1 viral load in the spinal cord and ganglions using RT-PCR. The results evidenced alteration of sensitivity in the animals from the 7th day that remained until the 28th day after inoculation of the virus, a change in the MAPKs proteins expression, with a raise of expression of JNK, AKT e CREB in the dorsal horn of the spinal cord in the 8th and 21st day after the HSV-1 virus inoculation, thus suggesting the participation of these proteins in the alteration of sensitivity both in the herpetic and post herpetic periods. It is also possible to observe the presence of viral load in the spinal cord and left spinal ganglion in the herpetic period after HSV-1 virus inoculation

Recentemente, estudos têm associado a presença de vírus da família herpesviridae à doença periodontal, os quais poderiam estar envolvidos na ocorrência e progressão de diferentes formas da doença periodontal, através da supressão do sistema imune do periodonto, liberação de citotoxinas, mediadores pró-inflamatórios, o que poderia favorecer o crescimento subgengival de microrganismos. Neste estudo, testamos a hipótese de que a prevalência do herpes vírus na placa subgengival de pacientes com gengivite é igual a de pacientes portadores de periodontite crônica. Desse modo, o presente estudo teve como objetivo determinar a presença dos vírus Herpes simples vírus tipo 1 (HSV-1), Citomegalovírus (HCMV) e Epstein-Barr vírus tipo 1 (EBV-1), relacionando-os com a presença de bactérias periodontopatogênicas como: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Tannerella forsythia (Tf) e Dialister pneumosintes (Dp) em amostras de placa subgengival, coletadas de 30 pacientes portadores de periodontite crônica (grupo PC), 30 pacientes com gengivite (grupo G) e 30 indivíduos periodontalmente saudáveis (grupo C). Foram coletadas quatro amostras de placa subgengival do sítio mais profundo de cada quadrante nos pacientes do grupo PC e nos pacientes do grupo G e C, um sítio aleatório de cada quadrante foi examinado. A detecção de espécies bacterianas e de herpes vírus na placa subgengival dos grupos foi realizada por PCR e Nested PCR, respectivamente. A análise estatística mostrou que o HCMV foi detectado com freqüência similar nos três grupos estudados e que houve uma maior prevalência do HSV-1, EBV-1 e P. intermedia nos pacientes com periodonite crônica e gengivite em relação ao grupo controle. Houve associação da periodontite crônica com o EBV-1 e as cinco bactérias estudadas, além da associação entre os vírus (EBV-1+ HCMV; EBV-1 + HSV-1; HSV-1 + HCMV) e entre vírus e bactérias (EBV-1+ P.intermedia, EBV-1 + P. gingivalis; HCMV + T. forsythia; HCMV + A. actinomycetemcomitans; HSV-1 + T. forsythia; HSV-1 + P. gingivalis). / Recently, studies have linked the presence of the virus family herpesviridae and periodontal disease, which may be involved in the occurrence and progression of different forms of periodontal disease through the suppression of the immune system of the periodontium, release of cytotoxins, pro-inflammatory mediators and immunopathological events, may promote growth of subgingival microorganisms. In this study, we tested the hypothesis that the prevalence of herpes virus in sub-gingival plaque of patients with gingivitis is equal to patients with chronic periodontitis. Thus, this study aimed to determine the presence of the virus Herpes simplex virus type 1 (HSV-1), Cytomegalovirus (HCMV) and Epstein-Barr virus type 1 (EBV-1), relating to the presence of bacteria as periodontopathogens: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Tannerella forsythia (Tf) and Dialister pneumosintes (Dp) in subgingival plaque samples collected from 30 patients with chronic periodontitis (CP group), 30 patients with gingivitis (group G) and 30 periodontally healthy subjects (group C). We collected four samples of subgingival plaque from the deepest site in each quadrant and in the PC group and patients in group C and G, a random site in each quadrant was examined. The detection of bacterial species and herpes virus in subgingival plaque of the groups were identified by PCR and nested PCR respectively. Statistical analysis showed that HCMV was detected with a similar frequency among the three groups and significant difference in prevalence of HSV-1, EBV-1 and P. intermedia in patients with gingivitis in the control group. The chronic periodontitis was associated with EBV-1 and the five bacteria studied, and the association of the virus (EBV-1 + HCMV, EBV-1 + HSV-1, HSV-1 + HCMV) and between viruses and bacteria (EBV-1+ P.intermedia, EBV-1 + P. gingivalis; HCMV + T. forsythia; HCMV + A. actinomycetemcomitans, HSV-1 + T. forsythia; HSV-1 + P. gingivalis).

A dor é descrita como uma sensação sensorial e emocional desagradável de extrema importância para sobrevivência e integridade do organismo. As dores crônicas de origem neuropática são de difícil tratamento e seus mecanismos fisiopatológicos pouco conhecidos. Este estudo foi realizado após inoculação do vírus HSV-1 na pata traseira esquerda de camundongos machos da linhagem Balb/C, e teve como objetivo investigar comportamentalmente o desenvolvimento de alodínia mecânica e hipernocicepção nas fases herpética e pós herpética, caracterizar a atividade de vias intracelulares de sinalização das proteínas JNK, AKT CREB, P38, ERK, Glutamina Sintetase e Stat 3, através de western blot na coluna dorsal da medula espinal nas fases herpética e pós herpética, além de verificar a presença do vírus HSV-1 na medula espinal e gânglio dos animais por meio da reação em cadeia da polimerase em tempo real (RT-PCR). Os resultados evidenciaram alteração de sensibilidade nos animais a partir do 7º dia que permaneceu até o 28º dia após a inoculação do vírus. Houve uma mudança na expressão das proteínas MAPKs, com aumento na expressão de JNK, AKT e CREB no corno dorsal da medula no 8º e 21º dia após a inoculação do vírus HSV-1, aumento na expressão de P 38 e P ERK no 8º dia após inoculação do vírus e uma diminuição na expressão da proteína Stat 3 no 8º e 21º dia após a inoculação do vírus, sugerindo assim a participação dessas proteínas na alteração de sensibilidade tanto no período herpético quanto pós herpético. Também ocorreu um aumento na amplificação do DNA viral HSV-1 na medula espinal e gânglio espinal esquerdo no período herpético após inoculação do vírus HSV-1 / The pain is described as a sensorial and emotional unpleasant sensation of extreme importance for survival and integrity of the organism. The chronic pains that has neuropathic source are hard to treat and its physiopathologic mechanisms not well known. This study was performed after inoculation of the virus HSV-1 in the left back foot of male Balb/C mouse, and had as main objectives to behaviorally investigate the development of mechanical allodynia and hyper nociception during the herpetic and post-herpetic phase, characterize the activity of the intracellular signalization paths of the proteins JNK, AKT CRB, P38, ERK, glutamine synthetase and Sat 3 during herpetic and post-herpetic phase using Western Blot, besides checking as well for the presence of HSV-1 viral load in the spinal cord and ganglions using RT-PCR. The results evidenced alteration of sensitivity in the animals from the 7th day that remained until the 28th day after inoculation of the virus, a change in the MAPKs proteins expression, with a raise of expression of JNK, AKT e CREB in the dorsal horn of the spinal cord in the 8th and 21st day after the HSV-1 virus inoculation, thus suggesting the participation of these proteins in the alteration of sensitivity both in the herpetic and post herpetic periods. It is also possible to observe the presence of viral load in the spinal cord and left spinal ganglion in the herpetic period after HSV-1 virus inoculation

Recentemente, estudos têm associado a presença de vírus da família herpesviridae à doença periodontal, os quais poderiam estar envolvidos na ocorrência e progressão de diferentes formas da doença periodontal, através da supressão do sistema imune do periodonto, liberação de citotoxinas, mediadores pró-inflamatórios, o que poderia favorecer o crescimento subgengival de microrganismos. Neste estudo, testamos a hipótese de que a prevalência do herpes vírus na placa subgengival de pacientes com gengivite é igual a de pacientes portadores de periodontite crônica. Desse modo, o presente estudo teve como objetivo determinar a presença dos vírus Herpes simples vírus tipo 1 (HSV-1), Citomegalovírus (HCMV) e Epstein-Barr vírus tipo 1 (EBV-1), relacionando-os com a presença de bactérias periodontopatogênicas como: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Tannerella forsythia (Tf) e Dialister pneumosintes (Dp) em amostras de placa subgengival, coletadas de 30 pacientes portadores de periodontite crônica (grupo PC), 30 pacientes com gengivite (grupo G) e 30 indivíduos periodontalmente saudáveis (grupo C). Foram coletadas quatro amostras de placa subgengival do sítio mais profundo de cada quadrante nos pacientes do grupo PC e nos pacientes do grupo G e C, um sítio aleatório de cada quadrante foi examinado. A detecção de espécies bacterianas e de herpes vírus na placa subgengival dos grupos foi realizada por PCR e Nested PCR, respectivamente. A análise estatística mostrou que o HCMV foi detectado com freqüência similar nos três grupos estudados e que houve uma maior prevalência do HSV-1, EBV-1 e P. intermedia nos pacientes com periodonite crônica e gengivite em relação ao grupo controle. Houve associação da periodontite crônica com o EBV-1 e as cinco bactérias estudadas, além da associação entre os vírus (EBV-1+ HCMV; EBV-1 + HSV-1; HSV-1 + HCMV) e entre vírus e bactérias (EBV-1+ P.intermedia, EBV-1 + P. gingivalis; HCMV + T. forsythia; HCMV + A. actinomycetemcomitans; HSV-1 + T. forsythia; HSV-1 + P. gingivalis). / Recently, studies have linked the presence of the virus family herpesviridae and periodontal disease, which may be involved in the occurrence and progression of different forms of periodontal disease through the suppression of the immune system of the periodontium, release of cytotoxins, pro-inflammatory mediators and immunopathological events, may promote growth of subgingival microorganisms. In this study, we tested the hypothesis that the prevalence of herpes virus in sub-gingival plaque of patients with gingivitis is equal to patients with chronic periodontitis. Thus, this study aimed to determine the presence of the virus Herpes simplex virus type 1 (HSV-1), Cytomegalovirus (HCMV) and Epstein-Barr virus type 1 (EBV-1), relating to the presence of bacteria as periodontopathogens: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Tannerella forsythia (Tf) and Dialister pneumosintes (Dp) in subgingival plaque samples collected from 30 patients with chronic periodontitis (CP group), 30 patients with gingivitis (group G) and 30 periodontally healthy subjects (group C). We collected four samples of subgingival plaque from the deepest site in each quadrant and in the PC group and patients in group C and G, a random site in each quadrant was examined. The detection of bacterial species and herpes virus in subgingival plaque of the groups were identified by PCR and nested PCR respectively. Statistical analysis showed that HCMV was detected with a similar frequency among the three groups and significant difference in prevalence of HSV-1, EBV-1 and P. intermedia in patients with gingivitis in the control group. The chronic periodontitis was associated with EBV-1 and the five bacteria studied, and the association of the virus (EBV-1 + HCMV, EBV-1 + HSV-1, HSV-1 + HCMV) and between viruses and bacteria (EBV-1+ P.intermedia, EBV-1 + P. gingivalis; HCMV + T. forsythia; HCMV + A. actinomycetemcomitans, HSV-1 + T. forsythia; HSV-1 + P. gingivalis).

Le virus de l'herpès simplex de type 1 (HSV1) établit une infection latente dans le système nerveux de l'homme, au cours de laquelle un type de transcrits, appelés LATs (pour latency associated transcripts), s'accumule dans les neurones infectés. Le rôle clef des LATs dans le contrôle de la latence virale est reconnu. Cependant, depuis leur découverte dans les années 80, leur mécanisme d'action reste non élucidé.Le gène des LATs est transcrit en un LAT primaire de 8,3kb, qui est épissé, conduisant à la formation de deux LATs stables : le LAT2kb et le LAT1.5kb. De façon remarquable, le LAT2kb et le LAT1.5kb sont des introns. Leur stabilité est la conséquence d'un branchement non canonique qui se traduit par le maintien de la structure en lariat. Par ailleurs, la région du génome codant les LATs contient également le gène RL2 qui code ICP0, la protéine la plus en amont dans la cascade de réactivation du virus. Des études précédentes ont montré qu'au moment de la latence, des transcrits RL2 non épissés, s'accumulent au site principal de la latence (le ganglion de Gasser).Nous avons caractérisé ces transcrits non épissés du gène RL2 dans les tissus infectés de façon latente. Ils contiennent de façon reproductible l'intron 1 et sont d'autant plus abondants dans les tissus infectés de façon latente que les LAT s'accumulent. On peut ainsi distinguer plusieurs types de tissus infectés de façon latente, dont les deux exemples les plus représentatifs sont d'une part le ganglion de Gasser (forte expression des LAT et accumulation de transcrits RL2 non-épissés) et d'autre part le ganglion cervical supérieur (pas d'accumulation de LAT par rapport aux quantités exprimées pendant la phase aiguë de l'infection, et très peu d'expression dans transcrits non-épissés). Dans tous les cas, la réalité du caractère latent de l'infection était confirmé par la présence de génome viral sans expression de transcrits matures de gène viral précoce (représenté par celui de la thymidine kinase) ni tardif (gène UL18). Ces résultats suggèrent une relation entre la présence des LAT et l'accumulation de transcrits RL2 non-épissés, ce qui pourrait être en relation avec le maintien de l'infection à l'état latent dans ces tissus.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Herpes simplex virus Typ 1 (HSV-1) ist ein humanpathogenes Virus der Familie Herpesviridae. Für eine erfolgreiche Virusreplikation besitzt HSV-1 mehrere Gene, die in den meisten infizierten Zelltypen Apoptose verhindern. Im Gegensatz dazu führt die HSV-1 Infektion eines zentralen Zelltyps des Immunsystems, den unreifen dendritischen Zellen (iDCs), zu Apoptose. Dies könnte ein Aspekt der HSV-1 Immunevasion sein. Bisher waren die Ursachen der Apoptose von HSV-1 infizierten iDCs unzureichend aufgeklärt. Es wurde jedoch gezeigt, dass das antiapoptotische zelluläre Protein c-FLIP in HSV-1 infizierten iDCs reduziert ist. In dieser Arbeit wurde die c-FLIP Menge in iDCs erstmalig mit Hilfe von RNA Interferenz erfolgreich reduziert. Dies bestätigte die Bedeutung von c-FLIP für die Lebensfähigkeit von iDCs. Folglich könnte auch die Reduktion der c-FLIP Menge nach HSV-1 Infektion iDCs für Apoptose empfindlich machen. Die HSV-1 induzierte c-FLIP Reduktion erfolgte in späten Stadien der Infektion, abhängig von der ordnungsgemäßen Expression viraler „early“ und „leaky late“ Gene. Sie fand nicht auf RNA Ebene statt und war unabhängig vom Proteasom und der Bindung an den „death inducing signaling complex“. Stattdessen wurde c-FLIP wahrscheinlich von einer viralen oder zellulären Protease abgebaut. In dieser Arbeit wurde erstmals gezeigt, dass zusätzlich zu Veränderungen im zellulären Apoptosesignalnetzwerk der Mangel an einem antiapoptotischen viralen Faktor zur Apoptose von HSV-1 infizierten iDCs beiträgt. Eine Microarray Analyse der HSV-1 Genexpression ergab, dass HSV-1 Latenz-assoziierte Transkripte (LATs) in apoptotischen iDCs signifikant geringer exprimiert waren als in nicht-apoptotischen epithelialen Zellen. LATs besitzen in Neuronen und epithelialen Zellen eine antiapoptotische Aktivität. Diese könnte den Mangel an c-FLIP kompensieren. Übereinstimmend mit dieser Hypothese induzierte eine HSV-1 LAT-Deletionsmutante mehr Apoptose in iDCs im Vergleich zum Wildtyp-Virus. / Herpes simplex virus type 1 (HSV-1) is a human pathogen which belongs to the family Herpesviridae. HSV-1 encodes several genes, which serve to efficiently prevent apoptosis in most infected cell types, thereby ensuring successful virus replication. In contrast, HSV-1 infection of one central cell type of the immune system, immature dendritic cells (iDCs), results in apoptosis. This could be one aspect of HSV-1 immunevasion. So far, the mechanisms underlying apoptosis of HSV-1 infected iDCs were poorly defined. However, it has been shown that the antiapoptotic cellular protein c-FLIP is reduced in HSV-1 infected iDCs. In this work, the amount of c-FLIP was for the first time successfully reduced in iDCs by RNA interference. This confirmed the importance of c-FLIP for viability of iDCs. Therefore, it is likely that c-FLIP reduction after HSV-1 infection also sensitizes iDCs to apoptosis. HSV-1 induced c-FLIP reduction occurred at late stages of infection and was dependent on proper expression of early and leaky late virus genes. Furthermore, it was not operative at the RNA level and was independent from the proteasome and binding to the death inducing signaling complex. Rather, c-FLIP was presumably degraded by a viral or cellular protease. In this work it was shown for the first time, that in addition to changes in the cellular apoptosis signaling network, the lack of one antiapoptotic viral factor contributes to apoptosis of HSV-1 infected iDCs. HSV-1 latency-associated transcripts (LATs) were significantly lower expressed in apoptotic iDCs compared to non-apoptotic epithelial cells, determined by microarray analysis of HSV-1 gene expression. It is known that in neurons and epithelial cells, LATs possess a potent antiapoptotic activity. This could compensate the lack of c-FLIP. Consistent with this hypothesis, a LAT deletion mutant of HSV-1 induced more apoptosis in iDCs compared to the respective wild type virus.

L'encéphalite herpétique (EH), causée par l'herpès simplex virus-1 (HSV-1), peut résulter de défauts monogéniques de l'immunité médiée par TLR3. L'induction d'interférons (IFNs)-α/β ou -λ via TLR3 est cruciale à la protection après infection primaire avec HSV-1 dans le système nerveux central (SNC). Nous décrivons deux enfants avec l'EH portant différentes mutations hétérozygotes (D50A et G159A) dans TBK1, encodant TANK-Binding Kinase 1, une kinase aux carrefours de multiples voies de signalisation induisant des IFNs. Les deux allèles mutants de TBK1 sont perte-de-fonction par des mécanismes différents: instabilité de la protéine (D50A) ou perte d'activité kinase (G159A). Ces allèles sont associés à un trait autosomal dominant (AD) par des mécanismes différents: haplotype-insuffisance (D50A) ou dominance négative (G159A). Un défaut de réponses à poly(I:C) par TLR3 est observable dans les fibroblastes hétérozygotes pour G159A, et non pour D50A TBK1. Néanmoins, la réplication virale et la mortalité cellulaire après infection par deux virus dépendants de TLR3 (HSV-1 et VSV) étaient élevées dans les fibroblastes des deux patients. Ces phénotypes peuvent être sauvés par IFN-α2b. De plus, la production d'IFNs en réponse à des agonistes et virus indépendants de TLR3 est maintenue dans les PBMCs et fibroblastes des patients. Le phénotype cellulaire restreint, partiel représente ainsi le phénotype clinique de ces patients, limité à l'EH. Ces données identifient la déficience partielle AD de TBK1 comme une nouvelle étiologie génétique de l'EH de l'enfance, et indiquent que TBK1 est essentiel pour le contrôle de HSV-1 dans le SNC, médié par TLR3 et dépendant des IFNs

The detection of immunostimulatory DNA is well documented to occur at several cellular sites, but there is limited evidence of nuclear innate DNA sensing. Prior to this study, the detection of herpesviral DNA was thought to be restricted to the cytosol so as to limit the sensing of host DNA in the nucleus. However, given the nuclear lifecycle of these viruses, we hypothesized that viral DNA could be sensed in the nucleus of infected cells. To test this hypothesis we examined the activation of interferon regulatory factor 3 (IRF-3) in response to herpes simplex virus 1 (HSV-1) infection of primary human foreskin fibroblasts (HFF). Using a mutant defective for expression of all viral genes, we observed that the release of viral DNA into the nucleus is necessary to activate IRF-3 signaling. Furthermore, we determined this response to be dependent on nuclear-localized interferon inducible protein 16 (IFI16) and the cytoplasmic stimulator of interferon genes (STING) adaptor protein.