rabbit polyclonal anti hsv 1 antibody  (Novus Biologicals)

Journal: mBio

Article Title: Caspase-mediated DDX46 cleavage unchains antiviral immunity

doi: 10.1128/mbio.03519-25

Figure Lengend Snippet: Virus infection or treatment with RNA ligands induces reduced DDX46 protein levels. ( A–C ) HeLa cells were mock-infected or infected with VSV ( A ), NDV ( B ), or HSV-1 ( C ) at an MOI of 1 for 6, 12, 18, and 24 h. Protein levels of VSV-G, NDV-NP, or HSV-1-gD were analyzed by WB. β-Actin served as the loading control. ( D–F ) Representative results, with graphs representing the band intensity ratios of DDX46/β-actin normalized to the control conditions for the VSV ( D ), NDV ( E ), and HSV-1 ( F ) infection groups. ( G–J ) HeLa cells were transfected with RNA ligands [poly(I:C) or 3p-hpRNA, panels G and H ] or DNA ligands [poly(G:C) or HSV-60, panels I and J ] for 18 h. Protein levels of DDX46 were analyzed by WB. β-Actin served as the loading control. Representative results, with graphs representing the band intensity ratios of DDX46/β-actin normalized to the control conditions for the RNA ligands ( H ) or DNA ligand treatment groups ( J ). ( K ) Schematic diagram of DDX46 isoform I (full-length; DDX46-I) and isoform II (lacking valine at amino acid 872; DDX46-II). ( L–N ) HeLa cells were transfected with the empty vector p3×Flag, Flag-DDX46-I, or Flag-DDX46-II for 24 h, then mock infected or infected with VSV ( L ), NDV ( M ), or HSV-1 ( N ) at an MOI of 1 for 6, 12, 18, and 24 h. Protein levels of exogenous Flag-DDX46 and viral proteins (VSV-G, NDV-NP, or HSV-1-gD) were analyzed by WB. β-Actin served as the loading control. Data are presented as means from three independent experiments. *** P < 0.001.

Article Snippet: The rabbit polyclonal anti-HSV-1 antibody (NB600-516) was obtained from Novus Biologicals.

Techniques: Virus, Infection, Control, Transfection, Plasmid Preparation

Journal: mBio

Article Title: Caspase-mediated DDX46 cleavage unchains antiviral immunity

doi: 10.1128/mbio.03519-25

Figure Lengend Snippet: Viral infection induces DDX46 cleavage and translocation from the nucleus to the cytoplasm. ( A–D ) HeLa cells were transfected with the Flag-DDX46 for 24 h, then mock-infected or infected with VSV ( A, B ) or HSV-1 ( C, D ) at an MOI of 1 for 6 and 12 h. Cells were fixed and subjected to IF analysis using anti-Flag and anti-viral-protein (VSV-G or HSV-1-gD) antibodies. Nuclei were counterstained with DAPI. Quantification of the relative percentages of cells with nuclear (Nuc) and cytoplasmic (Cyto) Flag signal after VSV ( B ) or HSV-1 ( D ) infection. Six randomly selected fields were analyzed using ImageJ. ( E ) Schematic of DDX46 mutants: point mutant D226A, N-terminal truncation (1–225), and C-terminal truncation (227–1,032). ( F, G ) HeLa cells were transfected with Flag-tagged WT-DDX46 or mutants (D226A, 1–225, and 227–1,032) for 24 h, then mock-infected or infected with VSV (MOI = 1) for 12 h. Cells were fixed and subjected to IF analysis using anti-Flag and anti-VSV-G antibodies. Nuclei were counterstained with DAPI. The two panels on the left show wider fields at 63×, and the two panels on the right show smaller fields at 20× ( F ). Quantification of the relative percentages of cells with nuclear or cytoplasmic Flag signal after VSV infection. Six randomly selected fields were analyzed using ImageJ ( G ). ( H and I ) HeLa cells were transfected with Flag-tagged WT-DDX46 or D226A-DDX46 for 24 h, then mock-infected ( H ) or infected with VSV at an MOI of 1 ( I ) for 12 h. Cells were harvested, and nuclear and cytoplasmic fractions were prepared using a nucleocytoplasmic isolation kit. Protein levels of exogenous Flag-DDX46 and VSV-G were analyzed by WB. β-Tubulin and Lamin B1 served as the loading controls for cytoplasmic and nuclear fractions, respectively. ( J ) Representative results, with graphs showing the band intensity ratios of DDX46/β-actin normalized to the control conditions for the VSV infection group ( I ). Data are presented as means from three independent experiments. *** P < 0.001.

Article Snippet: The rabbit polyclonal anti-HSV-1 antibody (NB600-516) was obtained from Novus Biologicals.

Techniques: Infection, Translocation Assay, Transfection, Mutagenesis, Isolation, Control

Journal: PLoS ONE

Article Title: Systemic CD8 + T Cell-Mediated Tumoricidal Effects by Intratumoral Treatment of Oncolytic Herpes Simplex Virus with the Agonistic Monoclonal Antibody for Murine Glucocorticoid-Induced Tumor Necrosis Factor Receptor

doi: 10.1371/journal.pone.0104669

Figure Lengend Snippet: CT26/NY-ESO-1 tumor sections from untreated mice (control) or mice injected i.t. with DTA-1, HF10, or HF10 combined with DTA-1 were stained with hematoxylin and eosin (A), and phycoerythrin (PE)-conjugated anti-CD8α mAb and DAPI (B). (C) Frozen sections of CT26/NY-ESO-1 tumors from mice i.t. injected with HF10 combined with DTA-1 were stained with a PE-anti-CD8α monoclonal antibody, a fluorescein isothiocyanate (FITC)-anti-HSV-1 polyclonal antibody, and DAPI. (D) CT26/NY-ESO-1 growth (mm 2 ) in both i.t. HF10- and DTA-1-treated control or CD8 + cell-depleted mice was measured. Seven mice per group were used. (E) Bilateral CT26/NY-ESO-1-bearing mice were treated with a combination of HF10 and DTA-1 in the tumors on the right flanks. Subsequent tumor growth (mm 2 ) of the treated right and contralateral left sites was measured. Tumor growth in untreated mice was measured and used as a control. Fourteen mice per group were used. By the Kruskal-Wallis ANOVA test, CT26/NY-ESO-1 growth inhibition by the combined HF10 and DTA-1 treatment in contralateral as well as treated sites was significantly different from the untreated control group. (F) CT26/NY-ESO-1 tumors from one side of bilateral tumor-bearing mice were treated i.t. with HF10 combined with DTA-1. Frozen sections of contralateral CT26/NY-ESO-1 tumors were stained with a PE-anti-CD8α monoclonal antibody, a fluorescein isothiocyanate (FITC)-anti-HSV-1 polyclonal antibody, and DAPI.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated and/or phycoerythrin (PE)-conjugated anti-mouse CD4 (RM4-5; eBioscience, Inc), anti-mouse CD8α mAb (53-6.7; BD Bioscience), anti-mouse/rat Foxp3 mAb (FJK-16s; eBioscience, Inc), anti-mouse IFN-γ mAb (XMG1.2; eBioscience, Inc), anti-mouse F4/80 mAb (BM8; BioLegend), and anti-rat IgG2b monoclonal antibodies (mAbs) (MRG2b-85; BioLegend) as well as a FITC-conjugated rabbit anti-HSV-1 polyclonal antibody (Dako) were used in flow cytometric analysis and immunohistochemstry.

Techniques: Injection, Staining, Inhibition

Journal: Molecular Therapy Oncolytics

Article Title: Efficacy of a third-generation oncolytic herpes simplex virus in refractory soft tissue sarcoma xenograft models

doi: 10.1016/j.omto.2022.04.010

Figure Lengend Snippet: Immunohistochemical analysis of HSV-1 in xenograft tumors Athymic mice were subcutaneously transplanted with SK-LMS-1 (A–D) and RMS-YM (E–H) cells, and the resulting subcutaneous tumors were inoculated with T-01 (2.0 × 10 6 PFU) or PBS twice weekly (days 0 and 3). Mice were euthanized on day 7 after inoculation, and tissue sections were stained with H&E (A, B, E, and F), anti-HSV-1 antibody (C, D), or X-gal (G, H). ICR mice with bilateral subcutaneous tumors arising from the CCRF S-180II cells (I–N) were established, and one of the bilateral subcutaneous tumors was inoculated with T-01 (2.0 × 10 6 PFU) or PBS twice weekly (days 0 and 3). Mice were euthanized 7 days after inoculation, and histological images of tissue sections stained with H&E (I–K) or X-gal (L–N) are shown. Representative images from these experiments are presented.

Article Snippet: The sections were treated to inhibit endogenous peroxidase activity and prevent nonspecific binding of the secondary antibody, incubated with a rabbit polyclonal anti-HSV-1 antibody (1:50,000) (Dako Cytomation), rinsed, and then incubated with an horseradish peroxidase-conjugated goat anti-rabbit immunoglobulin (Ig)G antibody (Nichirei Bioscience, Tokyo, Japan).

Techniques: Immunohistochemical staining, Staining