Additional file 1 of A new polysaccharide platform constructs self-adjuvant nanovaccines to enhance immune responses
Creators: name: Chen, Sisi
name: Yang, Liu
name: Ou, Xia
name: Li, Jin-Yu
name: Zi, Cheng-Ting
name: Wang, Hao
name: Hu, Jiang-Miao
name: Liu, Ye
Publisher: name: figshare
Year: 2022
Subjects: subject: Immunology
subject: FOS: Clinical medicine

description: Additional file 1: Table S1. Major chemical content of the polysaccharide PRBS. Table S2. The results of chemical analyses of PRBS fractions. Figure S1. Ultraviolet spectrum of PRBS (2 mg/mL) was analyzed by Shimadzu UV-2700 UV-vis spectrophotometer in the wavelength range of 190-600 nm. Figure S2. Polysaccharide purity by high-performance gel permeation chromatography (HPGPC) profiles. Figure S3. Standard curve of average molecular weight determination by high-performance gel permeation chromatography (HPGPC) profiles. Figure S4. HPLC analysis of monosaccharide standards and PRBS after PMP derivation. Figure S5. FT-IR spectra showing the characteristic absorption peaks of PRBS such as hydroxyl and carbony l.20 μL PRBS (2 mg/mL) aqueous solution mixed with KBr powder, dried and pressed, then detected by FT-IR. Table S3. Chemical shifts for the resonances of glycosyl residues of PRBS 1H /13C NMR spectra. Figure S6. TEM images of naked PRBS, RBD protein and HIV plasmid. Figure S7. (a) DLS exhibited that the hydrate size of PRBS-RBD protein nanovaccine on 1/4,1/2,1,2,3 days. (b) DLS exhibited that the hydrate size of PRBS-HIV plasmid nanovaccine on 1/4,1/2,1,2,3 days. Table S4. Hydrogen-bonds between PRBS and RBD. Figure S8. Intermolecular interactions of the PRBS docked to HIV plasmid. Figure S9. The inhibitory concentration curves of anti-SARS-CoV-2 neutralizating antibodies induced by either PRBS-RBD protein nanovaccine (10 μg RBD protein and 50 μg PRBS per each injection) or traditional RBD protein vaccine (10 μg RBD protein per each injection). Figure S10. IgG responses in three mouse vaccination groups: Blank group (100 μL saline per each injection), RBD vaccine (10 μg RBD protein per each injection), and 100 μL PRBS-RBD nanovaccine (10 μg RBD protein and 50 μg PRBS). All groups’ data from 1:10000 dilution. Figure S11. IgG subtype responses (IgG1, IgG2a, IgG2b, IgG3) in three mouse vaccination groups: Blank group (100 μL saline per each injection), RBD vaccine (10 μg RBD protein per each injection) and 100 μL PRBS-RBD nanovaccine (10 μg RBD protein and 50 μg PRBS). All groups’ data from 1:1000 dilution. All values are expressed as mean ± SEM for three duplicates. *P &lt; 0.05 versus the control group. Figure S12. The specific IFN-γ ELISPOT photography results of RBD-specific T cell response induced by either PRBS-RBD protein nanovaccine (10 μg RBD protein and 50 μg PRBS per each injection) or traditional RBD protein vaccine (10 μg RBD protein per each injection). Figure S13. The representative gating strategy between parallel groups. Figure S14. Western blotting results of HIV env plasmid expressing gp140 protein. Figure S15. The IC50 of anti-HIV neutralizing antibodies induced by either PRBS-HIV plasmid nanovaccine (10 μg HIV plasmid and 50 μg PRBS per each injection) or traditional HIV DNA vaccine (10 μg HIV plasmid per each injection). Figure S16. The specific IFN-γ ELISPOT photography results of HIV-specific T cell response induced by either PRBS-HIV plasmid nanovaccine (10 μg HIV plasmid and 50 μg PRBS per each injection) or traditional HIV DNA vaccine (10 μg HIV plasmid per each injection). Figure S17. The representative detailed gating strategy and parallel groups results from either PRBS-RBD protein nanovaccine group (10 μg RBD protein and 50 μg PRBS per each injection) or traditional RBD protein vaccine group (10 μg RBD protein per each injection). Figure S18. The representative detailed gating strategy and parallel groups results from either PRBS-HIV plasmid nanovaccine group (10 μg HIV plasmid and 50 μg PRBS per each injection) or traditional HIV plasmid vaccine group (10 μg HIV plasmid per each injection). Figure S19. Different concentrations of PRBS (100 μg/mL or 200 μg/mL) promote the phagocytosis ability of macrophages. Figure S20. Fluorescence images for the phagocytosis of dendritic cells (DC2.4). Figure S21. PRBS-specific IgG responses in two mouse vaccination groups: PRBS (50 μg PRBS per each injection), and negative control (100 μL salin per each injection). Figure S22. (a) IgG responses in two mouse vaccination groups: Alu-RBD vaccine (10 μg RBD protein and 35 μg Alu per each injection), and 100 μL PRBS-RBD nanovaccine (10 μg RBD protein and 50 μg PRBS per each injection). All groups’ data from 1:10000 dilution. (b) The titer of HIV-specific IgG induced by either CpG-HIV vaccine (15 μg CpG and 10 μg HIV plasmid per each injection) or 100 μL PRBS-HIV nanovaccine (10 μg HIV plasmid and 50 μg PRBS per each injection) (c) The magnitude of RBD-specific T cell response (IFN-γ ELISPOT) induced by either Alu-RBD vaccine (10 μg RBD protein and 35 μg Alu per each injection) or 100 μL PRBS-RBD nanovaccine (10 μg RBD protein and 50 μg PRBS per each injection). (d) The magnitude of HIV-specific T cell response (IFN-γ ELISPOT) induced by either CpG-HIV vaccine (15 μg CpG and 10 μg HIV plasmid per each injection) or 100 μL PRBS-HIV nanovaccine (10 μg HIV plasmid and 50 μg PRBS per each injection).