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wt hsv 1  (ATCC)


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    ATCC wt hsv 1
    Impaired induction of IFNs via TBK1-mediated pathways in the patient’s SV40-fibroblasts. (A) TBK1 mRNA levels (left panel) were measured by RT-qPCR in fibroblasts (SV40-fibroblasts) from healthy controls (C1, C2), P1, and two other TBK1-deficient patients (WT/G159A, W619*/W619*), and an immunoblot analysis of endogenous TBK1 protein levels (right panel) was performed with antibodies against the N-terminus and C-terminus of TBK1. The results shown are representative of three independent experiments. (B) SV40-fibroblasts from healthy controls (C1, C2), P1, two other TBK1 patients (WT/G159A, W619*/W619*), and a TLR3 −/− HSE patient were left unstimulated (NS) or were stimulated with poly (I:C) alone, Lipofectamine alone (Lipo), or both (poly (I:C)+Lipo), for 6 h. The relative expression levels of IFNB1 , IFNL1 , IFIT1 , and IL6 were measured by RT-qPCR. The results shown are from three independent experiments. (C) IFNB mRNA levels were measured by RT-qPCR in SV40-F from healthy controls ( n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies. Cells were either left uninfected (NS) or infected <t>with</t> <t>HSV-1</t> (KOS strain, MOI = 1) for 24 h. Data represent the means of three independent experiments. (D) SV40-F from healthy controls (Ctrls, n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies were either left untreated or pretreated with IFN-β for 24 h, followed by infection with HSV-1 (MOI = 0.001). Viral replication was assessed at the indicated time points post-infection using the TCID 50 virus titration method. Data represent means ± SEM from three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test; P values at 48 h after infection compare P1’s cells to controls. ***P < 0.0001. Source data are available for this figure: .
    Wt Hsv 1, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 193 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 193 article reviews
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    1) Product Images from "Recurrent severe viral infection in a child with inherited complete TBK1 deficiency"

    Article Title: Recurrent severe viral infection in a child with inherited complete TBK1 deficiency

    Journal: Journal of Human Immunity

    doi: 10.70962/jhi.20250058

    Impaired induction of IFNs via TBK1-mediated pathways in the patient’s SV40-fibroblasts. (A) TBK1 mRNA levels (left panel) were measured by RT-qPCR in fibroblasts (SV40-fibroblasts) from healthy controls (C1, C2), P1, and two other TBK1-deficient patients (WT/G159A, W619*/W619*), and an immunoblot analysis of endogenous TBK1 protein levels (right panel) was performed with antibodies against the N-terminus and C-terminus of TBK1. The results shown are representative of three independent experiments. (B) SV40-fibroblasts from healthy controls (C1, C2), P1, two other TBK1 patients (WT/G159A, W619*/W619*), and a TLR3 −/− HSE patient were left unstimulated (NS) or were stimulated with poly (I:C) alone, Lipofectamine alone (Lipo), or both (poly (I:C)+Lipo), for 6 h. The relative expression levels of IFNB1 , IFNL1 , IFIT1 , and IL6 were measured by RT-qPCR. The results shown are from three independent experiments. (C) IFNB mRNA levels were measured by RT-qPCR in SV40-F from healthy controls ( n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies. Cells were either left uninfected (NS) or infected with HSV-1 (KOS strain, MOI = 1) for 24 h. Data represent the means of three independent experiments. (D) SV40-F from healthy controls (Ctrls, n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies were either left untreated or pretreated with IFN-β for 24 h, followed by infection with HSV-1 (MOI = 0.001). Viral replication was assessed at the indicated time points post-infection using the TCID 50 virus titration method. Data represent means ± SEM from three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test; P values at 48 h after infection compare P1’s cells to controls. ***P < 0.0001. Source data are available for this figure: .
    Figure Legend Snippet: Impaired induction of IFNs via TBK1-mediated pathways in the patient’s SV40-fibroblasts. (A) TBK1 mRNA levels (left panel) were measured by RT-qPCR in fibroblasts (SV40-fibroblasts) from healthy controls (C1, C2), P1, and two other TBK1-deficient patients (WT/G159A, W619*/W619*), and an immunoblot analysis of endogenous TBK1 protein levels (right panel) was performed with antibodies against the N-terminus and C-terminus of TBK1. The results shown are representative of three independent experiments. (B) SV40-fibroblasts from healthy controls (C1, C2), P1, two other TBK1 patients (WT/G159A, W619*/W619*), and a TLR3 −/− HSE patient were left unstimulated (NS) or were stimulated with poly (I:C) alone, Lipofectamine alone (Lipo), or both (poly (I:C)+Lipo), for 6 h. The relative expression levels of IFNB1 , IFNL1 , IFIT1 , and IL6 were measured by RT-qPCR. The results shown are from three independent experiments. (C) IFNB mRNA levels were measured by RT-qPCR in SV40-F from healthy controls ( n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies. Cells were either left uninfected (NS) or infected with HSV-1 (KOS strain, MOI = 1) for 24 h. Data represent the means of three independent experiments. (D) SV40-F from healthy controls (Ctrls, n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies were either left untreated or pretreated with IFN-β for 24 h, followed by infection with HSV-1 (MOI = 0.001). Viral replication was assessed at the indicated time points post-infection using the TCID 50 virus titration method. Data represent means ± SEM from three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test; P values at 48 h after infection compare P1’s cells to controls. ***P < 0.0001. Source data are available for this figure: .

    Techniques Used: Quantitative RT-PCR, Western Blot, Expressing, Infection, Virus, Titration



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    Impaired induction of IFNs via TBK1-mediated pathways in the patient’s SV40-fibroblasts. (A) TBK1 mRNA levels (left panel) were measured by RT-qPCR in fibroblasts (SV40-fibroblasts) from healthy controls (C1, C2), P1, and two other TBK1-deficient patients (WT/G159A, W619*/W619*), and an immunoblot analysis of endogenous TBK1 protein levels (right panel) was performed with antibodies against the N-terminus and C-terminus of TBK1. The results shown are representative of three independent experiments. (B) SV40-fibroblasts from healthy controls (C1, C2), P1, two other TBK1 patients (WT/G159A, W619*/W619*), and a TLR3 −/− HSE patient were left unstimulated (NS) or were stimulated with poly (I:C) alone, Lipofectamine alone (Lipo), or both (poly (I:C)+Lipo), for 6 h. The relative expression levels of IFNB1 , IFNL1 , IFIT1 , and IL6 were measured by RT-qPCR. The results shown are from three independent experiments. (C) IFNB mRNA levels were measured by RT-qPCR in SV40-F from healthy controls ( n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies. Cells were either left uninfected (NS) or infected <t>with</t> <t>HSV-1</t> (KOS strain, MOI = 1) for 24 h. Data represent the means of three independent experiments. (D) SV40-F from healthy controls (Ctrls, n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies were either left untreated or pretreated with IFN-β for 24 h, followed by infection with HSV-1 (MOI = 0.001). Viral replication was assessed at the indicated time points post-infection using the TCID 50 virus titration method. Data represent means ± SEM from three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test; P values at 48 h after infection compare P1’s cells to controls. ***P < 0.0001. Source data are available for this figure: .
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    Impaired induction of IFNs via TBK1-mediated pathways in the patient’s SV40-fibroblasts. (A) TBK1 mRNA levels (left panel) were measured by RT-qPCR in fibroblasts (SV40-fibroblasts) from healthy controls (C1, C2), P1, and two other TBK1-deficient patients (WT/G159A, W619*/W619*), and an immunoblot analysis of endogenous TBK1 protein levels (right panel) was performed with antibodies against the N-terminus and C-terminus of TBK1. The results shown are representative of three independent experiments. (B) SV40-fibroblasts from healthy controls (C1, C2), P1, two other TBK1 patients (WT/G159A, W619*/W619*), and a TLR3 −/− HSE patient were left unstimulated (NS) or were stimulated with poly (I:C) alone, Lipofectamine alone (Lipo), or both (poly (I:C)+Lipo), for 6 h. The relative expression levels of IFNB1 , IFNL1 , IFIT1 , and IL6 were measured by RT-qPCR. The results shown are from three independent experiments. (C) IFNB mRNA levels were measured by RT-qPCR in SV40-F from healthy controls ( n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies. Cells were either left uninfected (NS) or infected <t>with</t> <t>HSV-1</t> (KOS strain, MOI = 1) for 24 h. Data represent the means of three independent experiments. (D) SV40-F from healthy controls (Ctrls, n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies were either left untreated or pretreated with IFN-β for 24 h, followed by infection with HSV-1 (MOI = 0.001). Viral replication was assessed at the indicated time points post-infection using the TCID 50 virus titration method. Data represent means ± SEM from three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test; P values at 48 h after infection compare P1’s cells to controls. ***P < 0.0001. Source data are available for this figure: .
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    Impaired induction of IFNs via TBK1-mediated pathways in the patient’s SV40-fibroblasts. (A) TBK1 mRNA levels (left panel) were measured by RT-qPCR in fibroblasts (SV40-fibroblasts) from healthy controls (C1, C2), P1, and two other TBK1-deficient patients (WT/G159A, W619*/W619*), and an immunoblot analysis of endogenous TBK1 protein levels (right panel) was performed with antibodies against the N-terminus and C-terminus of TBK1. The results shown are representative of three independent experiments. (B) SV40-fibroblasts from healthy controls (C1, C2), P1, two other TBK1 patients (WT/G159A, W619*/W619*), and a TLR3 −/− HSE patient were left unstimulated (NS) or were stimulated with poly (I:C) alone, Lipofectamine alone (Lipo), or both (poly (I:C)+Lipo), for 6 h. The relative expression levels of IFNB1 , IFNL1 , IFIT1 , and IL6 were measured by RT-qPCR. The results shown are from three independent experiments. (C) IFNB mRNA levels were measured by RT-qPCR in SV40-F from healthy controls ( n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies. Cells were either left uninfected (NS) or infected <t>with</t> <t>HSV-1</t> (KOS strain, MOI = 1) for 24 h. Data represent the means of three independent experiments. (D) SV40-F from healthy controls (Ctrls, n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies were either left untreated or pretreated with IFN-β for 24 h, followed by infection with HSV-1 (MOI = 0.001). Viral replication was assessed at the indicated time points post-infection using the TCID 50 virus titration method. Data represent means ± SEM from three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test; P values at 48 h after infection compare P1’s cells to controls. ***P < 0.0001. Source data are available for this figure: .
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    Impaired induction of IFNs via TBK1-mediated pathways in the patient’s SV40-fibroblasts. (A) TBK1 mRNA levels (left panel) were measured by RT-qPCR in fibroblasts (SV40-fibroblasts) from healthy controls (C1, C2), P1, and two other TBK1-deficient patients (WT/G159A, W619*/W619*), and an immunoblot analysis of endogenous TBK1 protein levels (right panel) was performed with antibodies against the N-terminus and C-terminus of TBK1. The results shown are representative of three independent experiments. (B) SV40-fibroblasts from healthy controls (C1, C2), P1, two other TBK1 patients (WT/G159A, W619*/W619*), and a TLR3 −/− HSE patient were left unstimulated (NS) or were stimulated with poly (I:C) alone, Lipofectamine alone (Lipo), or both (poly (I:C)+Lipo), for 6 h. The relative expression levels of IFNB1 , IFNL1 , IFIT1 , and IL6 were measured by RT-qPCR. The results shown are from three independent experiments. (C) IFNB mRNA levels were measured by RT-qPCR in SV40-F from healthy controls ( n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies. Cells were either left uninfected (NS) or infected <t>with</t> <t>HSV-1</t> (KOS strain, MOI = 1) for 24 h. Data represent the means of three independent experiments. (D) SV40-F from healthy controls (Ctrls, n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies were either left untreated or pretreated with IFN-β for 24 h, followed by infection with HSV-1 (MOI = 0.001). Viral replication was assessed at the indicated time points post-infection using the TCID 50 virus titration method. Data represent means ± SEM from three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test; P values at 48 h after infection compare P1’s cells to controls. ***P < 0.0001. Source data are available for this figure: .
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    Impaired induction of IFNs via TBK1-mediated pathways in the patient’s SV40-fibroblasts. (A) TBK1 mRNA levels (left panel) were measured by RT-qPCR in fibroblasts (SV40-fibroblasts) from healthy controls (C1, C2), P1, and two other TBK1-deficient patients (WT/G159A, W619*/W619*), and an immunoblot analysis of endogenous TBK1 protein levels (right panel) was performed with antibodies against the N-terminus and C-terminus of TBK1. The results shown are representative of three independent experiments. (B) SV40-fibroblasts from healthy controls (C1, C2), P1, two other TBK1 patients (WT/G159A, W619*/W619*), and a TLR3 −/− HSE patient were left unstimulated (NS) or were stimulated with poly (I:C) alone, Lipofectamine alone (Lipo), or both (poly (I:C)+Lipo), for 6 h. The relative expression levels of IFNB1 , IFNL1 , IFIT1 , and IL6 were measured by RT-qPCR. The results shown are from three independent experiments. (C) IFNB mRNA levels were measured by RT-qPCR in SV40-F from healthy controls ( n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies. Cells were either left uninfected (NS) or infected with HSV-1 (KOS strain, MOI = 1) for 24 h. Data represent the means of three independent experiments. (D) SV40-F from healthy controls (Ctrls, n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies were either left untreated or pretreated with IFN-β for 24 h, followed by infection with HSV-1 (MOI = 0.001). Viral replication was assessed at the indicated time points post-infection using the TCID 50 virus titration method. Data represent means ± SEM from three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test; P values at 48 h after infection compare P1’s cells to controls. ***P < 0.0001. Source data are available for this figure: .

    Journal: Journal of Human Immunity

    Article Title: Recurrent severe viral infection in a child with inherited complete TBK1 deficiency

    doi: 10.70962/jhi.20250058

    Figure Lengend Snippet: Impaired induction of IFNs via TBK1-mediated pathways in the patient’s SV40-fibroblasts. (A) TBK1 mRNA levels (left panel) were measured by RT-qPCR in fibroblasts (SV40-fibroblasts) from healthy controls (C1, C2), P1, and two other TBK1-deficient patients (WT/G159A, W619*/W619*), and an immunoblot analysis of endogenous TBK1 protein levels (right panel) was performed with antibodies against the N-terminus and C-terminus of TBK1. The results shown are representative of three independent experiments. (B) SV40-fibroblasts from healthy controls (C1, C2), P1, two other TBK1 patients (WT/G159A, W619*/W619*), and a TLR3 −/− HSE patient were left unstimulated (NS) or were stimulated with poly (I:C) alone, Lipofectamine alone (Lipo), or both (poly (I:C)+Lipo), for 6 h. The relative expression levels of IFNB1 , IFNL1 , IFIT1 , and IL6 were measured by RT-qPCR. The results shown are from three independent experiments. (C) IFNB mRNA levels were measured by RT-qPCR in SV40-F from healthy controls ( n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies. Cells were either left uninfected (NS) or infected with HSV-1 (KOS strain, MOI = 1) for 24 h. Data represent the means of three independent experiments. (D) SV40-F from healthy controls (Ctrls, n = 3), P1, and HSE patients with TBK1-deficient (WT/G159A) and AR IFNAR1 deficiencies were either left untreated or pretreated with IFN-β for 24 h, followed by infection with HSV-1 (MOI = 0.001). Viral replication was assessed at the indicated time points post-infection using the TCID 50 virus titration method. Data represent means ± SEM from three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test; P values at 48 h after infection compare P1’s cells to controls. ***P < 0.0001. Source data are available for this figure: .

    Article Snippet: SV40-immortalized fibroblasts (SV40-F; 5 × 10 4 cells/well) were seeded into 48-well plates and infected with WT HSV-1 (KOS strain, VR-1493; ATCC) at an MOI of 0.001.

    Techniques: Quantitative RT-PCR, Western Blot, Expressing, Infection, Virus, Titration