Journal: PLoS ONE

Article Title: Prevention of Herpes Simplex Virus Induced Stromal Keratitis by a Glycoprotein B-Specific Monoclonal Antibody

doi: 10.1371/journal.pone.0116800

Figure Lengend Snippet: Confluent Vero cells were infected with 100 TCID 50 of HSV-1 KOS and treated with either pooled human polyclonal HSV-neutralizing sera (1:40 in medium) or mAb 2c (500 nM). Cells were stained 48 h after infection for viral transmission with an HSV-1/2-glycoprotein D specific murine antibody and an Alexa488-conjugated secondary anti-mouse or anti-human antibody. Bound human antibodies or mAb 2c were detected with a Cy3-conjugated secondary antibody. Uninfected cells served as negative controls and showed no background staining (not shown). Magnification: 100x. Scale bar: 100 μm.

Article Snippet: HSV-1 infected cells were stained with a mouse anti-HSV-1/2-gD-antibody (Acris Antibodies, San Diego, CA, USA) and an Alexa Fluor 488 goat anti-mouse IgG specific secondary antibody (Invitrogen).

Techniques: Infection, Staining, Transmission Assay

Journal: PLoS ONE

Article Title: Prevention of Herpes Simplex Virus Induced Stromal Keratitis by a Glycoprotein B-Specific Monoclonal Antibody

doi: 10.1371/journal.pone.0116800

Figure Lengend Snippet: For the investigation of cell-to-neuron spread, epithelial C127I cells (arrows) were infected with the reporter virus HSV-1(17 + )Lox-Che. At 14 hpi cells were detached, treated with the indicated antibodies and co-cultured with uninfected DRG sensory neurons (arrowheads) (A-E) . For the investigation of neuron-to-cell transmission, DRG neurons were infected with HSV-1(17 + )Lox-Che. At 24 hpi, GFP-transfected, uninfected C127I were detached, treated with the indicated antibodies and co-cultured with the infected neurons (G-K) . For both settings, cells were fixed as indicated at 7, 22 and 30 hpi with PHEMO and labeled with anti-β-tubulin-III. Images were acquired with a confocal microscope. For the quantification of the cell-to-neuron (F) or neuron-to-cell transmission (L) , the values of mCherry-fluorescence were determined in a defined area within neurons (F) or GFP-positive C127I cells (G). (A-E and G-K) Scale bar is 10 µm. (F and L) . Error bars show standard error of the mean (SEM) of one representative experiment. Differences between the groups were statistically significant by a nonparametric ANOVA one way test (*** P < 0.001).

Article Snippet: HSV-1 infected cells were stained with a mouse anti-HSV-1/2-gD-antibody (Acris Antibodies, San Diego, CA, USA) and an Alexa Fluor 488 goat anti-mouse IgG specific secondary antibody (Invitrogen).

Techniques: Infection, Cell Culture, Transmission Assay, Transfection, Labeling, Microscopy, Fluorescence

Journal: PLoS ONE

Article Title: Prevention of Herpes Simplex Virus Induced Stromal Keratitis by a Glycoprotein B-Specific Monoclonal Antibody

doi: 10.1371/journal.pone.0116800

Figure Lengend Snippet: Lymphocytes derived from spleens or DLNs (R = ipsilateral, L = contralateral) of HSV-1 KOS infected mice receiving prophylactical or postexposure treatment (therapy) with mAb 2c were stimulated either with UV‑inactivated HSV-1 or Concavalin A for four days. Proliferation of CD4 + and CD8 + lymphocytes was determined by flow cytometry. Differences between the groups were statistically significant as calculated by a nonparametric ANOVA one way test (* P < 0.05; ** P < 0.01). Error bars represent the SEM.

Article Snippet: HSV-1 infected cells were stained with a mouse anti-HSV-1/2-gD-antibody (Acris Antibodies, San Diego, CA, USA) and an Alexa Fluor 488 goat anti-mouse IgG specific secondary antibody (Invitrogen).

Techniques: Derivative Assay, Infection, Flow Cytometry

Journal: PLoS ONE

Article Title: Prevention of Herpes Simplex Virus Induced Stromal Keratitis by a Glycoprotein B-Specific Monoclonal Antibody

doi: 10.1371/journal.pone.0116800

Figure Lengend Snippet: BALB/c mice (n = 5) were intravenously injected with 300 µg mAb 2c before corneal infection with HSV-1 KOS (1x105 PFU/cornea). At day 14 post infection, spleens and draining lymph nodes were isolated and homogenized. Cells were cultivated in triplicates in the presence of 2 × 10 7 PFU UV-inactivated HSV-1 or medium alone for stimulation. After 24 h, the contents of IL-2 and IFN-γ in cell supernatants were quantified by ELISA. Statistical analysis was undertaken with a nonparametric ANOVA test. Comparisons were considered significant at * P < 0.05. Error bars represent the standard error of the mean.

Article Snippet: HSV-1 infected cells were stained with a mouse anti-HSV-1/2-gD-antibody (Acris Antibodies, San Diego, CA, USA) and an Alexa Fluor 488 goat anti-mouse IgG specific secondary antibody (Invitrogen).

Techniques: Injection, Infection, Isolation, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Prevention of Herpes Simplex Virus Induced Stromal Keratitis by a Glycoprotein B-Specific Monoclonal Antibody

doi: 10.1371/journal.pone.0116800

Figure Lengend Snippet: (A) Percentage of reactivating ipsilateral or contralateral trigeminal ganglia from HSV‑1 KOS infected mice is shown. Trigeminal ganglia were isolated on day 14 after infection and co-cultured with confluent Vero cells for three weeks to investigate the reactivation of virus. The occurrence of a cytopathic effect was taken as proof of virus reactivation. Each group consisted of ten mice. The differences in the total numbers of reactivating virus between the ipsilateral and contralateral ganglia were examined with Fisher´s exact test. Comparisons were considered significant at * P < 0.05. (B) Subsequent investigation of the impact of prophylactic mAb 2c application at 24 h before infection on the frequency of reactivating virus after infection with decreasing viral loads. Each group contained 5 mice. (C) Quantification of HSV-1 DNA in trigeminal ganglia isolated on day 14 after infection. Each group contained 5 mice. Statistical analysis was undertaken with a nonparametric ANOVA test. Comparisons were considered significant at * P < 0.05. Error bars represent the standard error of the mean. I = ipsilateral; C = contralateral.

Article Snippet: HSV-1 infected cells were stained with a mouse anti-HSV-1/2-gD-antibody (Acris Antibodies, San Diego, CA, USA) and an Alexa Fluor 488 goat anti-mouse IgG specific secondary antibody (Invitrogen).

Techniques: Infection, Isolation, Cell Culture

Journal: PLoS ONE

Article Title: Prevention of Herpes Simplex Virus Induced Stromal Keratitis by a Glycoprotein B-Specific Monoclonal Antibody

doi: 10.1371/journal.pone.0116800

Figure Lengend Snippet: Mice were intravenously injected with 300µg of mAb hu2c (lower line) or PBS (upper line) at 48h after corneal infection with HSV-1 KOS. Six hours after injection, eyes were removed and sectioned. HSV-1 infection was stained with polyclonal goat anti-HSV-1 FITC conjugated antibodies and bound humanized antibody was detected with a Cy3-conjugated goat anti-human IgG secondary antibody. Nuclei were stained with Hoechst. Immunofluorescence images were acquired with a Zeiss Observer Z1 fluorescence microscope at a 200-fold magnification. Scale bars: 100 μm.

Article Snippet: HSV-1 infected cells were stained with a mouse anti-HSV-1/2-gD-antibody (Acris Antibodies, San Diego, CA, USA) and an Alexa Fluor 488 goat anti-mouse IgG specific secondary antibody (Invitrogen).

Techniques: Injection, Infection, Staining, Immunofluorescence, Fluorescence, Microscopy

Journal: Journal of Translational Autoimmunity

Article Title: Serologic markers of Epstein-Barr virus reactivation are associated with increased disease activity, inflammation, and interferon pathway activation in patients with systemic lupus erythematosus

doi: 10.1016/j.jtauto.2021.100117

Figure Lengend Snippet: Systemic lupus erythematosus (SLE) patients exhibit more frequent EBV reactivation compared with controls. Seropositivity for EBV-viral capsid antigen (VCA) IgG and IgA, EBV-early antigen (EA) IgG, CMV-IgG, and HSV-1 IgG were determined by ELISA for SLE patients (n = 175) and controls (n = 47). *p < 0.05, **p < 0.01 by two-tailed z-score test of population proportions.

Article Snippet: Antibodies against EBV-VCA (IgG and IgA), EBV-EA (IgG), cytomegalovirus (CMV, IgG), and herpes simplex virus-1 (HSV-1, IgG) were measured using commercial ELISAs (Wampole Laboratories) as previously described [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Journal of Translational Autoimmunity

Article Title: Serologic markers of Epstein-Barr virus reactivation are associated with increased disease activity, inflammation, and interferon pathway activation in patients with systemic lupus erythematosus

doi: 10.1016/j.jtauto.2021.100117

Figure Lengend Snippet: Systemic lupus erythematosus (SLE) patients exhibit higher levels of serological markers of Epstein-Barr virus (EBV) reactivation compared with controls. EBV-viral capsid antigen (VCA) IgG and IgA, EBV-early antigen (EA) IgG, cytomegalovirus (CMV) IgG, and herpes simplex virus (HSV-1) IgG antibody levels (international standardized ratio; ISR) were determined by ELISA for SLE patients (n = 175) and controls (n = 47). Each dot represents an independent sample, and data are represented as mean ± SD. *p < 0.05, **p < 0.01 by two-tailed student's t-test.

Article Snippet: Antibodies against EBV-VCA (IgG and IgA), EBV-EA (IgG), cytomegalovirus (CMV, IgG), and herpes simplex virus-1 (HSV-1, IgG) were measured using commercial ELISAs (Wampole Laboratories) as previously described [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Journal of Translational Autoimmunity

Article Title: Serologic markers of Epstein-Barr virus reactivation are associated with increased disease activity, inflammation, and interferon pathway activation in patients with systemic lupus erythematosus

doi: 10.1016/j.jtauto.2021.100117

Figure Lengend Snippet: Elevated concentrations of EBV-VCA (IgG and IgA) and EBV-EA (IgG) are associated with high SLE disease activity.

Article Snippet: Antibodies against EBV-VCA (IgG and IgA), EBV-EA (IgG), cytomegalovirus (CMV, IgG), and herpes simplex virus-1 (HSV-1, IgG) were measured using commercial ELISAs (Wampole Laboratories) as previously described [ ].

Techniques: Activity Assay

Journal: Journal of Translational Autoimmunity

Article Title: Serologic markers of Epstein-Barr virus reactivation are associated with increased disease activity, inflammation, and interferon pathway activation in patients with systemic lupus erythematosus

doi: 10.1016/j.jtauto.2021.100117

Figure Lengend Snippet: Seropositivity for Epstein-Barr virus (EBV)-early antigen (EA) IgG is associated with increased levels of EBV-related cytokine expression and type I IFN activity in systemic lupus erythematosus (SLE) patients. SLE patients were stratified based on seropositivity for (A–C) EBV-EA IgG (seropositive, n = 69; seronegative, n = 106) and (D) HSV-1 IgG (seropositive, n = 137; seronegative, n = 38). Plasma levels of (A) IL-10 and (B, D) IFN-induced protein 10 (IP-10) were determined using xMAP multiplex assays. Plasma levels of (C) B lymphocyte stimulator (BLyS) were determined by ELISA. Each dot represents an independent sample. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by two-tailed student's t-test.

Article Snippet: Antibodies against EBV-VCA (IgG and IgA), EBV-EA (IgG), cytomegalovirus (CMV, IgG), and herpes simplex virus-1 (HSV-1, IgG) were measured using commercial ELISAs (Wampole Laboratories) as previously described [ ].

Techniques: Expressing, Activity Assay, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Journal of Translational Autoimmunity

Article Title: Serologic markers of Epstein-Barr virus reactivation are associated with increased disease activity, inflammation, and interferon pathway activation in patients with systemic lupus erythematosus

doi: 10.1016/j.jtauto.2021.100117

Figure Lengend Snippet: Epstein-Barr virus (EBV)-early antigen (EA) IgG responses are enriched in previously defined systemic lupus erythematosus (SLE) patient clusters. SLE patients were stratified based on molecularly defined SLE disease clusters . Radar plots show SLEDAI scores and EBV-VCA IgG and IgA, EBV-EA IgG, CMV, and HSV-1 antibody levels in each patient cluster.

Article Snippet: Antibodies against EBV-VCA (IgG and IgA), EBV-EA (IgG), cytomegalovirus (CMV, IgG), and herpes simplex virus-1 (HSV-1, IgG) were measured using commercial ELISAs (Wampole Laboratories) as previously described [ ].

Techniques:

Journal: Frontiers in Genetics

Article Title: Development and Clinical Translation of Approved Gene Therapy Products for Genetic Disorders

doi: 10.3389/fgene.2019.00868

Figure Lengend Snippet: History and featured data of twenty approved gene and cell based gene therapy products.

Article Snippet: Imlygic , Amgen , USA FDA (2016) , Multiple solid tumors (Melanoma, Pancreatic Cancer) , HSV-1oncolytic virus with deletions in the γ34.5 and α47 regions, which GM-CSF gene inserted into the deleted γ34.5 loci. Tumor Lysis and induce antitumor immune responses , 10 6 -10 8 PFU/ml , Pregnancy, Immunocompromised Patients , $65000 per treatment , NCT02014441 NCT01740297 NCT02366195 NCT00402025 , .

Techniques: Inhibition, Injection, Plasmid Preparation, Infection, Blocking Assay, Variant Assay, Allele-specific Oligonucleotide, Lysis, Modification, Expressing, Transplantation Assay, Cell Counting