Journal: Advanced Science

Article Title: An Adenoviral‐Vectored Tp0326 Vaccine Elicits Robust Functional Antibodies to Prevent Treponema pallidum Dissemination in a Rabbit Model

doi: 10.1002/advs.78078

Figure Lengend Snippet: The immunological activity of serum antibodies in mice immunized with recombinant adenovirus and protein vaccines. (a) Flow chart of immunization plan for Balb/c Mice ( n = 5, respectively). (b) Detection of specific antibody titers at the immune endpoint in the serum of mice immunized with different vaccine groups. (c) The ratio of serum antibody IgG1 to IgG2a in each vaccine immunization group. (d) The antigen sensitivity of antibodies in each vaccine group and the detection of T.pallidum lysat. (e) Detection of the effects of antisera from each group on the adhesion of T.pallidum to cells by dark‐field microscopy. (f) Detection of the effects of different sera on the adhesion of T.pallidum to cells by indirect immunofluorescence. (g) Bactercidal of antibodies from different groups on the growth of T.pallidum cultivation in vitro. (h) The phagocytosis rate of each group was statistically analyzed according to the number of T.pallidum phagocytized by macrophage and separate tests were conducted on the Nichols reference strain and clinical strains. n.s.: no significance.

Article Snippet: ELISA kits for Mouse IgG1, Mouse IgG2a and Mouse IgG2b were purchased from Elabscience Co., Ltd. (China).

Techniques: Activity Assay, Recombinant, Vaccines, Sensitivity, Microscopy, Immunofluorescence, In Vitro

Journal: Journal of Fish Diseases

Article Title: Ocular Manifestations of Infectious Salmon Anaemia Virus Infection in Atlantic Salmon ( Salmo salar )

doi: 10.1111/jfd.70219

Figure Lengend Snippet: Antibody reagents used in this work.

Article Snippet: Mouse IgG2b , Mouse , Goat Polyclonal , Alkaline Phosphatase , Jackson Immunoresearch , 115‐055‐207.

Techniques: Cell-Signaling

Journal: Journal of Fish Diseases

Article Title: Ocular Manifestations of Infectious Salmon Anaemia Virus Infection in Atlantic Salmon ( Salmo salar )

doi: 10.1111/jfd.70219

Figure Lengend Snippet: Marker expression and morphological analysis of the central cornea region of ISAV‐infected Atlantic salmon. Immunohistochemistry for IgM expression showing the presence of IgM (red colour) in the central corneal epithelium (brackets) of non‐infected (A) and ISAV‐infected Atlantic salmon (B). Increased levels of IgM staining in the central corneal stroma of ISAV‐infected animals are shown in the boxed area. The characteristic morphological cellular layering of the central corneal epithelium indicated with black arrows in non‐infected Atlantic salmon (A) does not appear as prominant in the ISAV‐infected animals (B). A negative control for anti‐IgY‐AP is shown in the inset in (B). Immunohistochemistry for Sox6 showing negative results in non‐infected Atlantic salmon (C). Cells in the central corneal epithelium of ISAV‐infected Atlantic salmon were positive for Sox6 (D), with the highest signal being detected near Bowman's membrane (Bm). A negative control for anti‐IgG2b‐AP is shown in the inset in (D). Sox9 was negative within non‐infected Atlantic salmon (E) but positive in some cells in ISAV‐infected Atlantic salmon (F) and was localized near the outer central corneal epithelium (arrows in F). The upper right inset in (F) shows a magnified view of the cell boxed in black in (F). The bottom left inset in (F) shows a negative control on the same tissue for anti‐rabbit IgG‐AP. Immunohistochemistry for CD10 showed central corneal stromal staining in what appeared to be keratocytes in both non‐infected (G) and ISAV‐infected (H) salmon. A negative control on the same tissue for anti‐rabbit IgG‐AP is shown in the inset in (H). Giemsa and Herovichi histological stains highlighted the change in organization of the central corneal epithelium in healthy (I and K, bracket) versus ISAV‐infected (J, L) salmon. 400X. Scale bar subdivisions equal 10 μm.

Article Snippet: Mouse IgG2b , Mouse , Goat Polyclonal , Alkaline Phosphatase , Jackson Immunoresearch , 115‐055‐207.

Techniques: Marker, Expressing, Analysis, Infection, Immunohistochemistry, Staining, Negative Control, Membrane, Tissue

Journal: Journal of Fish Diseases

Article Title: Ocular Manifestations of Infectious Salmon Anaemia Virus Infection in Atlantic Salmon ( Salmo salar )

doi: 10.1111/jfd.70219

Figure Lengend Snippet: Marker expression and morphological analysis of the limbic region of cornea of ISAV‐infected Atlantic salmon. A, C, E, G, I, K, M, Corneal limbic region of non‐infected Atlantic salmon. B, D, F, H, J, L, N, Corneal limbic region of ISAV‐infected Atlantic salmon. A, B, Anti‐ISAV antigen IHC (red) indicating increased ISAV staining in cellular elements of the limbic corneal stroma associated with ISAV infection. C, D, Anti‐collagen type 1 IHC (red) indicating increased collagen type 1 staining in cellular elements of the limbic corneal stroma associated with ISAV infection. E, F, Giemsa, suggesting cellular infiltration in limbic corneal stroma of ISAV‐infected Atlantic salmon. G, H, Prussian blue, suggesting extracellular haemoglobin deposition (darker blue staining in black boxed area in H compared to G) in limbic corneal stroma and limbic corneal epithelial dysplasia in ISAV‐infected fish (white boxed area in H compared to G). I, J, Herovici, suggesting changes in collagen distribution patterns in limbic corneal stroma in ISAV‐infected Atlantic salmon. K, L, RGB‐trichrome, suggesting cellular infiltration and changes in extracellular matrix deposition patterns in limbic corneal stroma and limbic corneal epithelial dysplasia of ISAV‐infected Atlantic salmon (L), compared to uninfected (K). (M, N) Verhoeff elastin stain suggesting cellular infiltration, changes in extracellular matrix deposition patterns and limbic corneal epithelial dysplasia of ISAV‐infected salmon (N), compared to uninfected (M). Ce, corneal epithelium. Cs, corneal stroma. Cd, corneal dysplasia. Representative negative control for ISAV staining is shown in inset in B (cornea negative control for anti‐IgG2b‐AP). Representative negative control for Collagen 1 staining is shown in inset in D (cornea negative control for anti‐mouse IgG1‐AP). 400×. Scale bar subdivisions in M are 10 μm and representative for all panels.

Article Snippet: Mouse IgG2b , Mouse , Goat Polyclonal , Alkaline Phosphatase , Jackson Immunoresearch , 115‐055‐207.

Techniques: Marker, Expressing, Analysis, Infection, Staining, Negative Control

Journal: Journal of Fish Diseases

Article Title: Ocular Manifestations of Infectious Salmon Anaemia Virus Infection in Atlantic Salmon ( Salmo salar )

doi: 10.1111/jfd.70219

Figure Lengend Snippet: Marker expression and morphological analysis of the choroid rete mirabile in ISAV‐infected Atlantic salmon. (A, B) Representative ISAV antigen IHC showing (A), Vascular staining (red) for ISAV in the rete mirabile of a non‐infected Atlantic salmon and (B) Intense ISAV antigen staining in the vasculature (boxed) of an ISAV‐infected fish. Inset in (B) shows negative control staining using anti‐mouse IgG2b‐AP. (C) Giemsa staining of a non‐infected and (D) ISAV‐infected Atlantic salmon. Regions of cellular pooling are boxed. (E) RGB‐trichrome staining of a non‐infected and F, ISAV‐infected Atlantic salmon. Paucity of mature erythrocytes (green) and increased density of small round red/orange staining cells are evident across the whole image of the ISAV‐infected specimen. (G) Gata1 immunoreactivity of a non‐infected Atlantic salmon showing Gata1 staining (red) in fine capillary walls (arrows). (H) Gata1 immunoreactivity in ISAV‐infected fish showing staining (red) for both the vascular wall and numerous small round cells (exemplified by arrows in zoomed‐in area in left inset in H in which Gata1‐positive nucleoli are also visible). Inset on the right side in H shows negative control staining using anti‐mouse IgG1‐AP. All panels 400× except zoomed inset in (H). Scale bar subdivisions equal 10 μm.

Article Snippet: Mouse IgG2b , Mouse , Goat Polyclonal , Alkaline Phosphatase , Jackson Immunoresearch , 115‐055‐207.

Techniques: Marker, Expressing, Analysis, Infection, Staining, Negative Control

Journal: Journal of Fish Diseases

Article Title: Ocular Manifestations of Infectious Salmon Anaemia Virus Infection in Atlantic Salmon ( Salmo salar )

doi: 10.1111/jfd.70219

Figure Lengend Snippet: ISAV antigen expression and morphological analysis of the neural retina in ISAV‐infected Atlantic salmon. Representative IHC for ISAV antigen expression (red staining): No primary Ab negative control stained with anti‐mouse IgG2b‐AP only (A), non‐infected (B) and ISAV‐infected Atlantic salmon retina (C). Black boxed area in (C) indicates intense ISAV immunoreactivity in discrete regions of the inner retinal regions. (D) Giemsa staining of the non‐infected retina showing the retinal pigment epithelium (rpe), rods and cones (rc), outer nuclear layer (on), outer plexiform layer (op), inner nuclear layer (in), inner plexiform layer (ip), and ganglion cell layer/nerve fibre layer (gc). (E) Giemsa staining of Atlantic salmon infected with the pathogenic ISAV showing rosetting (black box) and scattering of the inner nuclear layer (white box) with rc, on, in and ip also indicated. 200×. Scale bar subdivisions equal 10 μm.

Article Snippet: Mouse IgG2b , Mouse , Goat Polyclonal , Alkaline Phosphatase , Jackson Immunoresearch , 115‐055‐207.

Techniques: Expressing, Analysis, Infection, Staining, Negative Control

Journal: Journal of Fish Diseases

Article Title: Ocular Manifestations of Infectious Salmon Anaemia Virus Infection in Atlantic Salmon ( Salmo salar )

doi: 10.1111/jfd.70219

Figure Lengend Snippet: CD10 expression and morphological analysis of extraocular muscle myofibrils of Atlantic salmon. Representative IHC for CD10 expression showing staining (red) in both the non‐infected (A) and ISAV‐infected (B) Atlantic salmon. Both striated (boxed) and punctate (arrows) staining was observed, with more intense staining seen in ISAV‐infected than non‐infected specimens. Inset on the right side of B shows a negative control stained using anti‐rabbit IgG‐AP. Herovici staining in non‐infected (C) and ISAV‐infected fish (D) is shown, with a reddish/brown staining seen in ISAV‐infected specimens. Muscle degeneration (D, arrows) and leukocytic‐like cell infiltration (D, boxed) are also observed in ISAV‐infected specimens. eom, extraocular muscle myofibril. 400×. Scale bar subdivisions equal 10 μm.

Article Snippet: Mouse IgG2b , Mouse , Goat Polyclonal , Alkaline Phosphatase , Jackson Immunoresearch , 115‐055‐207.

Techniques: Expressing, Analysis, Staining, Infection, Negative Control

Journal: Vaccines

Article Title: Exploring the Vaccine Adjuvant Effect and Mechanism of Epimedium Using Network Pharmacology, Molecular Docking, and Molecular Dynamics Simulations

doi: 10.3390/vaccines14050385

Figure Lengend Snippet: Flow chart showing the experimental design. The study consists of three phases: (1) Network pharmacology: screening of Epimedium bioactive compounds (TCMSP + SwissADME), target prediction (SwissTargetPrediction), immunomodulation-related genes collection (GeneCards/OMIM), PPI network construction (STRING), KEGG/GO enrichment, and “Epimedium-target-pathway-immunomodulation” network analysis. (2) Molecular docking and MD simulations: binding affinity evaluation and 100 ns dynamic simulation of ligand-target complexes. (3) In vivo validation: immunization of mice with HI antigen + ICA, measurement of HI-specific IgG subclasses by ELISA, and survival monitoring after lethal S. aureus USA300 challenge. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Subsequently, serum samples were added to the 96-well plate, serially diluted, and incubated for another 1 h. After washing the 96-well plate with PBST solution, 100 μL per well of horseradish peroxidase-conjugated goat anti-mouse IgG, IgG1, IgG2a, and IgG2b (Bethyl Biotech) were added and incubated at 37 °C for 1 h. Next, 100 μL per well of the corresponding enzyme substrate (Tian Gen) was added and allowed to react in a 37 °C incubator for 10 min.

Techniques: Binding Assay, In Vivo, Biomarker Discovery, Enzyme-linked Immunosorbent Assay

Journal: Vaccines

Article Title: Exploring the Vaccine Adjuvant Effect and Mechanism of Epimedium Using Network Pharmacology, Molecular Docking, and Molecular Dynamics Simulations

doi: 10.3390/vaccines14050385

Figure Lengend Snippet: Effects of ICA on HI-specific antibody levels and protection against S. aureus USA300 challenge in mice. ( A – E ) HI-specific IgG, IgG1, IgG2a, IgG2b, and the IgG1/IgG2a ratio in serum were measured by ELISA. ( F ) Survival rates of mice challenged with USA300 were monitored daily for 5 days. Data are presented as mean ± SD ( n = 8 per group). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Subsequently, serum samples were added to the 96-well plate, serially diluted, and incubated for another 1 h. After washing the 96-well plate with PBST solution, 100 μL per well of horseradish peroxidase-conjugated goat anti-mouse IgG, IgG1, IgG2a, and IgG2b (Bethyl Biotech) were added and incubated at 37 °C for 1 h. Next, 100 μL per well of the corresponding enzyme substrate (Tian Gen) was added and allowed to react in a 37 °C incubator for 10 min.

Techniques: Enzyme-linked Immunosorbent Assay