Journal: PLoS ONE

Article Title: Norovirus P Particle Efficiently Elicits Innate, Humoral and Cellular Immunity

doi: 10.1371/journal.pone.0063269

Figure Lengend Snippet: VA387 VLPs, P particles, P dimers were administrated to the mice (30 µg/mouse, n = 5) intranasally without adjuvant for three times in a two-week interval. Mice administered with PBS were regarded as negative controls (Ctrl). Sera were collected from each mouse prior to the first and one week after the final immunization. The titers of NoV-specific total IgG (A) and its subtypes IgG1 (B), IgG2a (C) and IgG2b (D) were determined by ELISA against purified P dimer. The antibody titers were defined as the endpoint dilution with a cut off signal intensity of 0.2. ∗ P <0.05 and ∗∗ P <0.01 were calculated vs. naïve mice control (Ctrl), while $ P <0.05 and $$ P <0.01 were calculated vs. VLP and # P <0.05 vs. P particle, respectively. Experiments were repeated three times.

Article Snippet: Baculovirus-expressed VLP of NoV strain VA387 (GII.4) was obtained from LigoCyte Pharmaceuticals.

Techniques: Enzyme-linked Immunosorbent Assay, Purification

Journal: PLoS ONE

Article Title: Norovirus P Particle Efficiently Elicits Innate, Humoral and Cellular Immunity

doi: 10.1371/journal.pone.0063269

Figure Lengend Snippet: (A) One week after the third immunization, splenocytes were collected and resuspended in RPMI-1640 with 3 µg/ml of brefeldin A and stimulated with 5 µg/ml of VA387 CD4 T cell epitope. Intracellular cytokines of IL-2, IFN-γ and TNF-α were stained as described . (B) Splenocytes were collected from immunized mice and stimulated with VA387-specific CD4 + T cell epitope without brefeldin A. IFN-γ productions in culture supernatant 24 or 48 hours post stimulation were determined by ELISA. ∗ P <0.01 and ∗∗ P <0.001 were calculated vs. naïve mice control (Ctrl), while $ P <0.01 and $$ P <0.001 were calculated vs. VLP and # P <0.05 and ## P <0.01 vs. P particle, respectively. Experiments were repeated three times.

Article Snippet: Baculovirus-expressed VLP of NoV strain VA387 (GII.4) was obtained from LigoCyte Pharmaceuticals.

Techniques: Staining, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Norovirus P Particle Efficiently Elicits Innate, Humoral and Cellular Immunity

doi: 10.1371/journal.pone.0063269

Figure Lengend Snippet: Splenocytes were isolated from immunized mice and labeled with carboxyfluorescein succinimidyl ester (CFSE). At the same time, mature bone marrow-derived DCs (BMDCs) pulsed with related antigens of VA387 VLPs, P particles, or P dimers were collected. Then, CFSE-labeled splenocytes were co-incubated with antigen-induced mature BMDCs for 5 days. Finally, cells were stained with APC-conjugated CD4 and were acquired by BD FACSCanto or BD Accuri™ C6. The resulting flow data were analyzed by FlowJo or Accuri™ C6 software. (A) Representative flow data of CD4 + T cell proliferation. (B) Percentages of proliferating CD4 + T cells (CD4 + CFSE low population) were shown in bar graph. ∗ P <0.001 and ∗∗ P <0.0001 were calculated vs. VLP, while $ P <0.001 and $$ P <0.0001 were calculated vs. P particle. Experiments were repeated three times.

Article Snippet: Baculovirus-expressed VLP of NoV strain VA387 (GII.4) was obtained from LigoCyte Pharmaceuticals.

Techniques: Isolation, Labeling, Derivative Assay, Incubation, Staining, Software