Journal: Journal of Biomedical Research

Article Title: NS3 epitope-decorated nanoparticles produced in bacteria trigger potent T cell immunity against hepatitis C virus

doi: 10.7555/JBR.39.20250197

Figure Lengend Snippet: Structure of recombinant proteins. A: Schematic representation of recombinant fusion proteins. B and C: Analysis of the amino acid sequence of the Sp (B) and Spn (C) linkers using the PCOILS program. The probability of formation of a coiled-coil structure is shown. Abbreviations: His, hexahistidine tag; NS3, epitopes from nonstructural protein 3; NS4a, epitopes from nonstructural protein 4A; NS5a, epitopes from nonstructural protein 5A; P, Th epitope PADRE; SAP, self-assembling peptide; Sp, helical linker; Spn, modified helical linker.

Article Snippet: Splenocytes were stimulated by incubation with 1 μg/mL of the PepTivator® HCV1b NS3 multiepitope peptide mixture (Miltenyi Biotec) for 54 h. Splenocyte proliferation was analyzed using the CCK-8 kit according to the manufacturer's instructions.

Techniques: Recombinant, Sequencing, Modification

Journal: Journal of Biomedical Research

Article Title: NS3 epitope-decorated nanoparticles produced in bacteria trigger potent T cell immunity against hepatitis C virus

doi: 10.7555/JBR.39.20250197

Figure Lengend Snippet: Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of recombinant proteins. A: Expression and purification of recombinant proteins. Lanes 1–3, His-SAP-Sp-P-NS3 (cell lysate, insoluble fraction, purified protein); lanes 4–6, His-SAP-Spn-P-NS3 (cell lysate, insoluble fraction, purified protein); lanes 7–9, His-SAP-Spn-P-NS3,5a,4a (cell lysate, insoluble fraction, purified protein); lane 10, molecular weight marker. B: Analysis of purified proteins with alkylation. The positions of the monomers are marked with an asterisk. Lane 1, His-SAP-Sp-P-NS3; lane 2, His-SAP-Spn-P-NS3; lane 3, His-SAP-Spn-P-NS3,5a,4a; lane 4, His-P-NS3,5a,4a.

Article Snippet: Splenocytes were stimulated by incubation with 1 μg/mL of the PepTivator® HCV1b NS3 multiepitope peptide mixture (Miltenyi Biotec) for 54 h. Splenocyte proliferation was analyzed using the CCK-8 kit according to the manufacturer's instructions.

Techniques: Polyacrylamide Gel Electrophoresis, SDS Page, Recombinant, Expressing, Purification, Molecular Weight, Marker

Journal: Journal of Biomedical Research

Article Title: NS3 epitope-decorated nanoparticles produced in bacteria trigger potent T cell immunity against hepatitis C virus

doi: 10.7555/JBR.39.20250197

Figure Lengend Snippet: Nanoparticle size distribution in purified protein samples revealed by dynamic light scattering analysis. Proteins: His-SAP-Sp-P-NS3 (A), His-SAP-Spn-P-NS3 (B), His-SAP-Spn-P-NS3,5a,4a (C), and His-P-NS3,5a,4a (D).

Article Snippet: Splenocytes were stimulated by incubation with 1 μg/mL of the PepTivator® HCV1b NS3 multiepitope peptide mixture (Miltenyi Biotec) for 54 h. Splenocyte proliferation was analyzed using the CCK-8 kit according to the manufacturer's instructions.

Techniques: Purification

Journal: Journal of Biomedical Research

Article Title: NS3 epitope-decorated nanoparticles produced in bacteria trigger potent T cell immunity against hepatitis C virus

doi: 10.7555/JBR.39.20250197

Figure Lengend Snippet: Analysis of immune response in vitro . Human dendritic cells were activated by recombinant proteins in vitro and then used to stimulate autologous lymphocytes. Stimulated lymphocytes were analyzed after a secondary stimulation with a mixture of peptides from the NS3 protein of the hepatitis C virus. A: Proliferative activity of lymphocytes. B–D: Interferon-γ (IFN-γ; B), interleukin (IL)-2 (C), and IL-4 (D) levels in the culture medium. Lanes: 1, PBS (negative control); 2, His-SAP-Sp; 3, His-SAP-Spn; 4, His-SAP-Sp-P-NS3; 5, His-SAP-Spn-P-NS3; 6, His-SAP-Spn-P-NS3,5a,4a; 7, His-P-NS3,5a,4a; 8, phytohemagglutinin (PHA, positive control). Each test was performed in triplicate. Data are shown as mean ± standard error of the mean. Normally distributed data were compared using the Student's t -test. * P < 0.05.

Article Snippet: Splenocytes were stimulated by incubation with 1 μg/mL of the PepTivator® HCV1b NS3 multiepitope peptide mixture (Miltenyi Biotec) for 54 h. Splenocyte proliferation was analyzed using the CCK-8 kit according to the manufacturer's instructions.

Techniques: In Vitro, Recombinant, Virus, Activity Assay, Negative Control, Positive Control

Journal: Journal of Biomedical Research

Article Title: NS3 epitope-decorated nanoparticles produced in bacteria trigger potent T cell immunity against hepatitis C virus

doi: 10.7555/JBR.39.20250197

Figure Lengend Snippet: Proliferative activity of splenocytes of immunized mice after stimulation with NS3 peptides. Lanes: 1, PBS (negative control); 2, His-SAP-Sp; 3, His-SAP-Spn; 4, His-SAP-Sp-P-NS3; 5, His-SAP-Spn-P-NS3; 6, His-SAP-Spn-P-NS3,5a,4а; 7, His-P-NS3,5a,4a; 8, phytohemagglutinin (PHA, positive control). Each test was performed in triplicate. Data are shown as mean ± standard error of the mean. Normally distributed data were compared using the Student's t -test. * P < 0.05.

Article Snippet: Splenocytes were stimulated by incubation with 1 μg/mL of the PepTivator® HCV1b NS3 multiepitope peptide mixture (Miltenyi Biotec) for 54 h. Splenocyte proliferation was analyzed using the CCK-8 kit according to the manufacturer's instructions.

Techniques: Activity Assay, Negative Control, Positive Control