Journal: ACS Applied Materials & Interfaces

Article Title: Assembly and Immunological Processing of Polyelectrolyte Multilayers Composed of Antigens and Adjuvants

doi: 10.1021/acsami.6b06275

Figure Lengend Snippet: Antigens and adjuvants can be assembled into iPEMs in a pH-dependent manner. (a) Quantification of final film thickness following deposition of eight bilayers of iPEMs composed of SIIN/polyIC or SIIN*/polyIC on silicon substrates. Thicknesses were measured by ellipsometry at different pH conditions for peptide, wash, and polyIC solutions. SIIN represents SIINFEKL and SIIN* represents SIINFEKL-R 9 . (b) Stepwise film growth of iPEMs composed of SIIN*/polyIC as a function of the pH of deposition solutions. (c) Representative absorbance traces demonstrating increasing antigen and adjuvant loading during deposition of iPEMs on quartz substrates using SIIN* (pH 8), wash (pH 8), and polyIC (pH 10) solutions. SIIN* (dotted green line, 508 nm); polyIC (dotted red line, 260 nm). (d) Relative loading of SIIN* and polyIC based on UV/vis measurements. Values represent the mean ± s.e.m. ( n = 5). Concentrations of antigen and adjuvant in deposition solutions were 0.2 mg/mL in (a) and 1.0 mg/mL in panels b–d.

Article Snippet: SIINFEKL (SIIN) and SIIINFEKL-R 9 (SIIN*) were synthesized by Genscript with >98% purity, with or without a FITC label on the N-terminus.

Techniques: Adjuvant

Journal: ACS Applied Materials & Interfaces

Article Title: Assembly and Immunological Processing of Polyelectrolyte Multilayers Composed of Antigens and Adjuvants

doi: 10.1021/acsami.6b06275

Figure Lengend Snippet: Effect of pH on iPEM assembly, diameter, and cargo loading. (a) Diameters and cargo loading of iPEMs assembled on sacrificial templates using different pH conditions for peptide solutions (PBS with 0.5 M NaCl), wash buffers (PBS with 0.05 M NaCl), and polyIC solutions (PBS with 0.5 M NaCl). Values represent the mean ± s.e.m., SIIN* represents SIINFEKL-R 9 . (b) Representative confocal microscopy images of iPEM-coated particles assembled using different pH conditions. Exposure times were fixed for all samples. (c) Diameter of iPEM-coated templates during assembly. d) Antigen and adjuvant loading in iPEM capsules after removal of the template and washing.

Article Snippet: SIINFEKL (SIIN) and SIIINFEKL-R 9 (SIIN*) were synthesized by Genscript with >98% purity, with or without a FITC label on the N-terminus.

Techniques: Confocal Microscopy, Adjuvant, Capsules

Journal: ACS Applied Materials & Interfaces

Article Title: Assembly and Immunological Processing of Polyelectrolyte Multilayers Composed of Antigens and Adjuvants

doi: 10.1021/acsami.6b06275

Figure Lengend Snippet: iPEM capsules are internalized by primary DCs with a high frequency of codelivery. (a) Representative histograms demonstrating SIIN* and polyIC uptake during treatment of splenic DCs with (SIIN*/polyIC) 3 iPEM capsules for 24 h. SIIN* represents SIINFEKL-R 9 . (b) Distribution of iPEM components in cells, calculated among cells positive for at least one iPEM signal (red, polyIC; green, SIIN*; blue, polyIC + SIIN* + ) to demonstrate the high degree of codelivery. (c) Viability of DCs treated with iPEM capsules relative to cells treated with LPS following a 24 h incubation. (d) Confocal microscopy images of DCs indicating internalization and colocalization of iPEM capsule components after a 2 h incubation (white, membrane; green, SIIN*; blue, nucleus; red, polyIC; scale bar = 20 μm). Values for all panels indicate the mean ± s.e.m.

Article Snippet: SIINFEKL (SIIN) and SIIINFEKL-R 9 (SIIN*) were synthesized by Genscript with >98% purity, with or without a FITC label on the N-terminus.

Techniques: Capsules, Incubation, Confocal Microscopy, Membrane

Journal: ACS Applied Materials & Interfaces

Article Title: Assembly and Immunological Processing of Polyelectrolyte Multilayers Composed of Antigens and Adjuvants

doi: 10.1021/acsami.6b06275

Figure Lengend Snippet: iPEMs activate DCs and promote antigen presentation. Expression of CD40 (a), CD80 (b), and CD86 (c) activation markers following treatment of primary DCs with iPEM capsules or the indicated controls for 24 h. SIIN represents SIINFEKL. d) Presentation of the SIINFEKL epitope through the MHC-I pathway assessed using anti-H-2k b -SIINFEKL following a 24 h incubation of primary DCs with iPEM capsules. Values for all panels indicate the mean ± s.e.m. Statistical comparisons indicate significance of **** = p ≤ 0.0001, and for clarity, are shown for each group compared against the control (Untreated; dashed line).

Article Snippet: SIINFEKL (SIIN) and SIIINFEKL-R 9 (SIIN*) were synthesized by Genscript with >98% purity, with or without a FITC label on the N-terminus.

Techniques: Immunopeptidomics, Expressing, Activation Assay, Capsules, Incubation, Control

Journal: ACS Applied Materials & Interfaces

Article Title: Assembly and Immunological Processing of Polyelectrolyte Multilayers Composed of Antigens and Adjuvants

doi: 10.1021/acsami.6b06275

Figure Lengend Snippet: iPEM capsules induce T cell proliferation and effector cytokine secretion in primary cell coculture. (a) Expansion of SIINFEKL-specific T cells following coculture with DCs treated with iPEM capsules for 24 h. Fluorescently labeled OT-I CD8 + T cells were isolated from spleens and cultured with DCs for 48 h, then proliferation was quantified by fluorescence dilution using flow cytometry. SIIN represents SIINFEKL. (b) Secretion of IFN-γ by the cells in panel a measured by ICCS. Values for all panels indicate the mean ± s.e.m. Statistical comparisons indicate significance of ** = p ≤ 0.01; **** = p ≤ 0.0001, and for clarity, are shown for each group compared against the control (Untreated; dashed line).

Article Snippet: SIINFEKL (SIIN) and SIIINFEKL-R 9 (SIIN*) were synthesized by Genscript with >98% purity, with or without a FITC label on the N-terminus.

Techniques: Capsules, Labeling, Isolation, Cell Culture, Fluorescence, Flow Cytometry, Control

Journal: ACS Applied Materials & Interfaces

Article Title: Assembly and Immunological Processing of Polyelectrolyte Multilayers Composed of Antigens and Adjuvants

doi: 10.1021/acsami.6b06275

Figure Lengend Snippet: Immunization with iPEM capsules promotes enhanced recall response of antigen-specific CD8 + T cells with successive immunization. C57BL/6 mice were immunized with equivalent doses of SIINFEKL and PolyIC formulated in iPEM capsules or using a free mixture at day 0, followed by booster injections at day 15 and 28 (red arrows). SIIN represents SIINFEKL. (a) Development of SIIN-specific tetramer kinetics over 35 days. (b) Representative flow cytometry plot from each treatment group in panel (a) shown on day 21 and day 35. (c) Tetramer frequencies of individual animals at day 35 among CD8 + T cells. (d) Rate of increase (slope) in antigen-specific CD8 + T cells occurring 1 week after each booster injection are greater following immunization with iPEMs relative to equivalent soluble vaccines. Slopes were calculated by measuring tetramer frequencies just before boosting, and again after 7 days. Values for panels (a-c) indicate the mean ± s.e.m. Statistical comparisons the end of the study (day 35) are indicated by * = p ≤ 0.05, *** = p ≤ 0.001, and **** = p < 0.0001.

Article Snippet: SIINFEKL (SIIN) and SIIINFEKL-R 9 (SIIN*) were synthesized by Genscript with >98% purity, with or without a FITC label on the N-terminus.

Techniques: Capsules, Flow Cytometry, Injection, Vaccines

Journal: ACS Biomaterials Science & Engineering

Article Title: In Vivo Expansion of Melanoma-Specific T Cells Using Microneedle Arrays Coated with Immune-Polyelectrolyte Multilayers

doi: 10.1021/acsbiomaterials.6b00414

Figure Lengend Snippet: Increased DCs activation after incubation with released MN cargo drives proliferation of Trp2-specific transgenic CD8 + T cells and effector cytokine secretion during coculture. (A) MFI of CFSE and (B) frequency of the CD8 + T cells as measured by flow cytometry. (C) Representative flow cytometry histograms showing the gating scheme and frequency of T cell proliferation. (D) IFN-γ, (E) IL-6, and (F) IL-1β concentrations in supernatants from the cocultures in A, B, and C measured by ELISA. Controls are vehicle (PBS), positive control (Trp2), negative controls (CpG, LPS), and irrelevant antigen control (SIIN).

Article Snippet: Trp2 180–188 (SVYDFFVWL), Trp2* (SVYDFFVWLRRRRRRRRR), and SIIN 257–264 (SIINFEKL) peptide from OVA protein were synthesized by Genscript with >98% purity, with or without a FITC label on the N-terminus.

Techniques: Activation Assay, Incubation, Transgenic Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Positive Control, Control

Journal: STAR Protocols

Article Title: Isolation and transcriptional profiling of antigen-presenting cells from mouse lung and MLNs following intranasal immunization

doi: 10.1016/j.xpro.2024.103400

Figure Lengend Snippet:

Article Snippet: SIIN-Q11 (Ac-SIINFEKL-GGAAY-QQKFQFQFEQQCONH2) , GenScript , N/A.

Techniques: Recombinant, Lysis, Software, Fluorescence

Journal: STAR Protocols

Article Title: Isolation and transcriptional profiling of antigen-presenting cells from mouse lung and MLNs following intranasal immunization

doi: 10.1016/j.xpro.2024.103400

Figure Lengend Snippet: The components of nanofiber solution

Article Snippet: SIIN-Q11 (Ac-SIINFEKL-GGAAY-QQKFQFQFEQQCONH2) , GenScript , N/A.

Techniques: Concentration Assay