Journal: Biomacromolecules

Article Title: Mannosylated Polycations Target CD206 + Antigen-Presenting Cells and Mediate T-Cell-Specific Activation in Cancer Vaccination

doi: 10.1021/acs.biomac.2c00993

Figure Lengend Snippet: CD86 expression and H-2K b –SIINFEKL presentation on JAWS II mouse DCs. CD86 (A,B) or H-2K b –SIINFEKL (C,D) positive cells percentage (A,C) and MFI (B,D) after cell incubation with pOVA, PEI L25kDa , M 58 A 45 , M 29 A 29 B 9 , and M 62 A 52 B 32 copolymers and PEI L25kDa /pOVA formulated at the N/P ratio of 10, and M 58 A 45 /pOVA, M 29 A 29 B 9 /pOVA, and M 62 A 52 B 32 /pOVA GPPs formulated at the N/P ratio of 2.5 (2.5 μg mL –1 pOVA concentration) as detected by flow cytometry. Untreated cells were used as control (Ctrl). (D) Negative values were considered as zero and labeled with a “+”. (E) CD206-mediated endocytosis of copolymer/pOVA complexes leads to increased CD86 expression and presentation of SIINFEKL antigenic peptide, derived from OVA, on MHC class I. Data are presented as mean ± s.d. ( n = 2). Statistical significance is reported as samples vs pOVA; * p < 0.05; ** p < 0.01; **** p < 0.0001.

Article Snippet: CMV-promoted pOVA was supplied by GeneScript (Leiden, Netherlands) and amplified by Escherichia coli bacterial cell transformation using One Shot TOP10 chemically competent E. coli (Invitrogen, Carlsbad, USA) and purified with a NucleoBond Xtra Midi kit (Macherey-Nagel GmbH, Düren, Germany). pDNA concentration and purity were measured on a NanoDrop One (ThermoFisher, UK) prior to complexation.

Techniques: Expressing, Incubation, Concentration Assay, Flow Cytometry, Control, Labeling, Derivative Assay

Journal: Biomacromolecules

Article Title: Mannosylated Polycations Target CD206 + Antigen-Presenting Cells and Mediate T-Cell-Specific Activation in Cancer Vaccination

doi: 10.1021/acs.biomac.2c00993

Figure Lengend Snippet: In vivo antitumor therapeutic efficacy. (A) Timeline for B16-OVA/B16-F1 tumor cell inoculation in C57BL/6 mice and schedule of treatment. (B) B16-OVA melanoma tumor growth after subcutaneous injection of pOVA (black), M 58 A 45 /pOVA (green), or M 62 A 52 B 32 /pOVA (red) ( N = 4/group). Control copolymer/pEGFP polyplexes are reported in Figure S20 . (C–E) Immunological analysis of T-cell population in B16-OVA tumor mass. T-cells were analyzed for their expression (reported as percentage of positive cells) of CD4 + (C) or CD8 + (D) molecules on the surface and for their ability to specifically recognize SIINFEKL bound to the MHC I molecule (E). (F) IFN-γ production by splenic T-cells isolated from B16-OVA tumor-bearing mice after their stimulation with SIINFEKL (blue bars) or gp100 (yellow bars) peptides, as revealed by ELISPOT assay. Results are reported as mean ± SEM ( n = 4, * p < 0.05, ** p < 0.01, **** p < 0.0001). FC: flow cytometry.

Article Snippet: CMV-promoted pOVA was supplied by GeneScript (Leiden, Netherlands) and amplified by Escherichia coli bacterial cell transformation using One Shot TOP10 chemically competent E. coli (Invitrogen, Carlsbad, USA) and purified with a NucleoBond Xtra Midi kit (Macherey-Nagel GmbH, Düren, Germany). pDNA concentration and purity were measured on a NanoDrop One (ThermoFisher, UK) prior to complexation.

Techniques: In Vivo, Drug discovery, Injection, Control, Expressing, Isolation, Enzyme-linked Immunospot, Flow Cytometry