Journal: Scientific Reports

Article Title: Induction of systemic and mucosal immune response against Zika virus by vaccination with non-infectious chimeric VLPs

doi: 10.1038/s41598-025-07059-6

Figure Lengend Snippet: Expression and assembly of VSP-VLPs ZIKV-E. ( A ) Fluorescence microscopy was conducted after transfecting VSP-expressing cells with the DNA constructs pZIKV-E and/or pGag. The expression of each specific protein is indicated at the top of each image, using the corresponding fluorescence color. Cells were analyzed for the expression of ZIKV-E and VSP proteins after incubating with the respective specific antibodies, followed by a fluorophore-conjugated secondary antibody. The expression of MLV-Gag was visualized by the fluorescence emitted from eYFP. In all fluorescence images, nuclei are labeled with DAPI (blue), and the yellow signal indicates co-expression. ( B ) Purified VSP-VLPs ZIKV-E were examined through transmission electron microscopy (TEM) and immunogold labeling, which confirmed the proper display of ZIKV-E (left), VSP (center) and MLV-Gag (right). Specific monoclonal antibodies were employed to identify the various components present in the particles. Immunogold-positive signals (black dots) corresponding to the specific markers are indicated by blue arrows.

Article Snippet: Then, fixed VSP-VLPs ZIKV-E were incubated with the anti-ZIKV GpE mAb (R&D Systems, MAB10009, 1/100), 7F5 mAb (1/100), and R187 mAb (ATCC ® CRL-1912TM, 1/1000).

Techniques: Expressing, Fluorescence, Microscopy, Construct, Labeling, Purification, Transmission Assay, Electron Microscopy, Bioprocessing

Journal: Scientific Reports

Article Title: Induction of systemic and mucosal immune response against Zika virus by vaccination with non-infectious chimeric VLPs

doi: 10.1038/s41598-025-07059-6

Figure Lengend Snippet: Detection of ZIKV-E from purified VSP-VLPs by Western blot. Purified VSP-VLPs ZIKV-E and VSP-VLPs without ZIKV-E (Ctrl) were resolved by SDS-PAGE, under reducing (5% β-ME) conditions, followed by immunodetection using anti-Zika envelope-specific antibody. The protein molecular weight marker (MW) is indicated on the left. The upper band (~ 58 kDa, red arrow) corresponds to the full-length ZIKV-E protein. A lower band (blue arrow), also recognised by the antibody, is marked as a putative truncated or degraded form of ZIKV-E.

Article Snippet: Then, fixed VSP-VLPs ZIKV-E were incubated with the anti-ZIKV GpE mAb (R&D Systems, MAB10009, 1/100), 7F5 mAb (1/100), and R187 mAb (ATCC ® CRL-1912TM, 1/1000).

Techniques: Purification, Western Blot, SDS Page, Immunodetection, Molecular Weight, Marker

Journal: Scientific Reports

Article Title: Induction of systemic and mucosal immune response against Zika virus by vaccination with non-infectious chimeric VLPs

doi: 10.1038/s41598-025-07059-6

Figure Lengend Snippet: Induction of anti-ZIKV systemic humoral response in mice vaccinated with VSP-VLPs ZIKV-E. ( A ) Mice were vaccinated orally or subcutaneously with VSP-VLPs ZIKV-E and the specific anti-ZIKV IgG plus IgM were determined in sera 14 days after each dose. ( B ) Serum IgG plus IgM titer was evaluated at 42 days post immunization (d.p.i.) ( C ) Serum levels of IgG1 subclass were measured at 42 d.p.i. ( D ) Serum levels of IgG2a subclass were measured at 42 d.p.i. ( E ) Serum IgA levels were assessed at 42 d.p.i. n = 6 from two independent experiments for VSP-VLPs ZIKV-E vaccinated groups; n = 5 for mock group. Data are shown as means ± SEM. In the antibodies kinetic response ( A ) data were analyzed by two-way ANOVA and Bonferroni’s multiple comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 respect to mock group at the same d.p.i.

Article Snippet: Then, fixed VSP-VLPs ZIKV-E were incubated with the anti-ZIKV GpE mAb (R&D Systems, MAB10009, 1/100), 7F5 mAb (1/100), and R187 mAb (ATCC ® CRL-1912TM, 1/1000).

Techniques:

Journal: Scientific Reports

Article Title: Induction of systemic and mucosal immune response against Zika virus by vaccination with non-infectious chimeric VLPs

doi: 10.1038/s41598-025-07059-6

Figure Lengend Snippet: Specific anti-ZIKV mucosal response after VSP-VLPs ZIKV-E immunization. Mice were vaccinated orally or subcutaneously with VSP-VLPs ZIKV-E and the specific anti-ZIKV antibodies were determined in mucosal samples 14 days after the last dose (42 d.p.i.). ( A ) IgG + IgM levels in vaginal swabs were measured. ( B ) Specific IgA levels in vaginal swabs. ( C ) Specific IgA levels in faecal samples. n = 6 from two independent experiments for VSP-VLPs ZIKV-E vaccinated groups; n = 5 for mock group. Data are shown as means ± SEM. * p < 0.05, ** p < 0.01.

Article Snippet: Then, fixed VSP-VLPs ZIKV-E were incubated with the anti-ZIKV GpE mAb (R&D Systems, MAB10009, 1/100), 7F5 mAb (1/100), and R187 mAb (ATCC ® CRL-1912TM, 1/1000).

Techniques:

Journal: Scientific Reports

Article Title: Induction of systemic and mucosal immune response against Zika virus by vaccination with non-infectious chimeric VLPs

doi: 10.1038/s41598-025-07059-6

Figure Lengend Snippet: Induction of anti-ZIKV systemic cellular immune response in mice vaccinated with VSP-VLPs ZIKV-E. Balb/c mice were vaccinated orally or subcutaneously with VSP-VLPs ZIKV-E and the cellular immune response against ZIKV was evaluated 14 days after the last dose (42 d.p.i.) by the measurement of cytokines in splenocyte supernatants. n = 6 from two independent experiments for VSP-VLPs ZIKV-E vaccinated groups; n = 5 for mock group. Data are shown as means ± SEM. * p < 0.05, ** p < 0.01.

Article Snippet: Then, fixed VSP-VLPs ZIKV-E were incubated with the anti-ZIKV GpE mAb (R&D Systems, MAB10009, 1/100), 7F5 mAb (1/100), and R187 mAb (ATCC ® CRL-1912TM, 1/1000).

Techniques:

Journal: Scientific Reports

Article Title: Induction of systemic and mucosal immune response against Zika virus by vaccination with non-infectious chimeric VLPs

doi: 10.1038/s41598-025-07059-6

Figure Lengend Snippet: VSP-VLPs ZIKV-E immunization increases neutralizing antibodies. Balb/c mice were vaccinated orally or subcutaneously at days 0, 14 and 28 with VSP-VLPs ZIKV-E. Neutralizing antibody endpoint titers were determined in serum samples collected 14 days after the final dose using ZIKV-AR. The neutralization titer was defined as the serum dilution that reduces the cytopathic effect by 50% (MNT50) ( A ) or 80% (MNT80) ( B ). A total of n = 6 animals were analyzed from two independent experiments in the VSP-VLPs ZIKV-E vaccinated groups, while n = 5 animals were analyzed from the mock group. Data are shown as means ± SEM. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Then, fixed VSP-VLPs ZIKV-E were incubated with the anti-ZIKV GpE mAb (R&D Systems, MAB10009, 1/100), 7F5 mAb (1/100), and R187 mAb (ATCC ® CRL-1912TM, 1/1000).

Techniques: Neutralization