Journal: Human Vaccines & Immunotherapeutics

Article Title: Emerging heterologous mRNA-based booster strategies within the COVID-19 vaccine landscape

doi: 10.1080/21645515.2022.2153532

Figure Lengend Snippet: A summary of the COVID-19 vaccine landscape as of 21 September 2022.

Article Snippet: Overall, 40 COVID-19 vaccines have been authorized worldwide, including those mentioned previously as well as inactivated virus whole-cell vaccines BBIBP-CorV (Covilo; Sinopharm, Beijing, China), BBV152 (Covaxin®; Bharat Biotech, Turakapally, India), and CoronaVac (Sinovac; Beijing, China); the non-replication viral vector Sputnik V (Gamaleya; Moscow, Russia), Ad5-nCoV (Convideia; CanSinoBio, Tianjin, China); and protein subunit-based vaccine COVOVAX TM (Serum Institute of India, Novovax formulation, Pune, India) (summarized in )., As of the same date, approximately 67% of the vaccine-eligible population has been fully vaccinated (received the primary vaccination-series) in the United States, among whom the majority received mRNA-based vaccines (38% received mRNA-1273 and 59% received BNT162b2).

Techniques: Plasmid Preparation, Recombinant

Journal: Cell reports

Article Title: Mannose-binding lectin and complement mediate follicular localization and enhanced immunogenicity of diverse protein nanoparticle immunogens

doi: 10.1016/j.celrep.2021.110217

Figure Lengend Snippet: (A) BLI analysis of eOD-GT8 60mer glycan variants binding to immobilized recombinant murine MBL2 as a function of eOD particle concentration. (B–D) C57Bl/6 mice (n = 5/group) were immunized with 2 μg eOD equivalent eOD-GT8 60mer glycan variants and saponin adjuvant. Shown are average intensity Z projections through 360 μm of cleared draining lymph nodes harvested on day 7 (B, blue, CD35; red, eOD-GT8 60mer; scale bars denote 500 μm), and analyses of normalized total eOD-GT8 60mer signal per Z plane of cleared lymph nodes (C) and percent eOD-60mer signal found within follicles (D). Error bars indicate SEM; points represent average values between paired draining lymph nodes from one animal; *, p <0.05; ***, p <0.001; ****, p <0.0001, ns = not significant by one-way ANOVA followed by Tukey post hoc test.

Article Snippet: Streptavidin-coated sensors were incubated in PBS containing 1% BSA and 0.1 M CaCl and were then loaded into wells of the same solution containing 1 μg/mL biotinylated murine MBL2 (R&D Systems 2208-MB-050/CF) for 1 min.

Techniques: Glycoproteomics, Binding Assay, Recombinant, Concentration Assay, Adjuvant

Journal: Cell reports

Article Title: Mannose-binding lectin and complement mediate follicular localization and enhanced immunogenicity of diverse protein nanoparticle immunogens

doi: 10.1016/j.celrep.2021.110217

Figure Lengend Snippet: (A and B) BLI binding curves of unmodified and PNGase F-treated HPV16 L1 (A) or unmodified HBsAg (B) to immobilized recombinant murine MBL2 as functions of antigen concentration. (C) C57Bl/6 mice or MBL KO mice (n = 5/group) were immunized with 0.1 μg AlexaFluor 647-labeled HPV16 L1 and saponin adjuvant. Seven days later, lymph nodes were harvested, cleared, and imaged by confocal microscopy. Shown are average intensity Z projections through 360 μm of tissue; shown is staining for CD35 (blue) and antigen (red), scale bars denote 500 μm. (D) Serum HPV16 L1-specific IgG titers over time in mice (n = 5/group) immunized with 0.1 μg HPV16 L1 and saponin adjuvant. Error bars indicate SEM, p = 0.92 compared with WT one-way ANOVA. (E) Absolute counts of germinal center B cells (B220 + GL7 + CD4 − CD38 low ) and antigen-specific germinal center B cells (B220 + GL7 + HPV16 L1 + CD4 − CD38 low ) from WT and MBL KO mice (n = 5/group) at day 12 following immunization with 0.1 μg HPV16 L1 and saponin adjuvant. Error bars indicate SEM; *, p < 0.05 by Mann-Whitney test. (F) C57BL/6 mice or MBL KO mice (n = 5/group) were immunized with 5 μg AlexaFluor 647-labeled HBsAg and saponin adjuvant. Seven days later, lymph nodes were harvested, cleared, and imaged by confocal microscopy. Shown are average intensity Z projections through 360 μm of tissue; shown is staining for CD35 (blue) and antigen (red), scale bars denote 500 μm. (G) Serum HBsAg-specific IgG titers over time in mice immunized with 5 μg HBsAg and saponin adjuvant. Error bars indicate SEM; *, p <0.05 compared with WT by one-way ANOVA followed by Tukey post hoc test. (H) Absolute counts of germinal center B cells (B220 + GL7 + CD4 − CD38 low ) and antigen-specific germinal center B cells (B220 + GL7 + HBsAg + CD4 − CD38 low ) from WT and C3 KO mice 12 days after immunization with 5 μg HBsAg and saponin adjuvant. Error bars indicate SEM; *, p <0.05 by Mann-Whitney test.

Article Snippet: Streptavidin-coated sensors were incubated in PBS containing 1% BSA and 0.1 M CaCl and were then loaded into wells of the same solution containing 1 μg/mL biotinylated murine MBL2 (R&D Systems 2208-MB-050/CF) for 1 min.

Techniques: Binding Assay, Recombinant, Concentration Assay, Labeling, Adjuvant, Confocal Microscopy, Staining, MANN-WHITNEY

Journal: Cell reports

Article Title: Mannose-binding lectin and complement mediate follicular localization and enhanced immunogenicity of diverse protein nanoparticle immunogens

doi: 10.1016/j.celrep.2021.110217

Figure Lengend Snippet: (A) Design models of glycosylated I53–50 nanoparticles with either 240 glycans (left) or 120 glycans (right) displayed on the particle. The left particle is assembled with 20 glycosylated I53–50A trimeric subunits (protein in gray and glycans in green) and 12 non-glycosylated I53–50B pentameric subunits (orange) to display 240 glycans on the particle. The right particle is assembled with 10 glycosylated and 10 non-glycosylated I53–50A trimeric subunits, and 12 non-glycosylated I53–50B pentameric subunits to display 120 glycans. The two-component nature of I53–50 particles (i.e., each particle is composed of 20 trimers and 12 pentamers) enabled titration of glycan densities on the particle through varying the molar ratio of non-glycosylated to glycosylated I53–50A trimeric subunits; glycosylation of I53–50A trimers was either native or high-mannose for each particle formulation. (B) Mean glycan distances calculated from the particle structure for NPs with titrated levels of total glycans. (C) BLI analysis of serially glycosylated I53–50 nanoparticles binding to immobilized recombinant murine MBL2 as a function of I53–50 nanoparticle concentration. (D) The apparent dissociation constant (K D ) of immobilized murine MBL2 binding to each I53–50 high-mannose glycoform was determined by BLI analysis using a global 1:1 binding model applied to the three highest I53–50 concentrations. (E and F) C57Bl/6 mice (n = 5/group) were immunized with 5 μg I53–50 high-mannose glycan variants and saponin adjuvant. Shown are average intensity Z projections through 360 μm of cleared draining lymph nodes harvested on days 3 and 7 (E, blue, CD35; red, I53–50; scale bars denote 500 μm), and quantification of the percent I53–50 signal found within follicles (F). Error bars indicate SEM; points represent average values between paired draining lymph nodes from one animal; *, p <0.05; ****, p <0.0001, ns = not significant by one-way ANOVA followed by Tukey post hoc test. (G) Absolute counts of germinal center B cells and antigen-specific germinal center B cells from WT and MBL KO mice 12 days after immunization with 5 μg I53–50 and saponin adjuvant. Error bars indicate SEM; *, p <0.05; ns = not significant by one-way ANOVA followed by Tukey post hoc test.

Article Snippet: Streptavidin-coated sensors were incubated in PBS containing 1% BSA and 0.1 M CaCl and were then loaded into wells of the same solution containing 1 μg/mL biotinylated murine MBL2 (R&D Systems 2208-MB-050/CF) for 1 min.

Techniques: Titration, Glycoproteomics, Formulation, Binding Assay, Recombinant, Concentration Assay, Adjuvant

Journal: Cell reports

Article Title: Mannose-binding lectin and complement mediate follicular localization and enhanced immunogenicity of diverse protein nanoparticle immunogens

doi: 10.1016/j.celrep.2021.110217

Figure Lengend Snippet:

Article Snippet: Streptavidin-coated sensors were incubated in PBS containing 1% BSA and 0.1 M CaCl and were then loaded into wells of the same solution containing 1 μg/mL biotinylated murine MBL2 (R&D Systems 2208-MB-050/CF) for 1 min.

Techniques: Recombinant, Staining, Antibody Labeling, Software