Journal: ACS Nano

Article Title: Combination mRNA Vaccine Adjuvanted with CpG Oligodeoxynucleotides Enhances Protection against Respiratory Virus Infection

doi: 10.1021/acsnano.5c14408

Figure Lengend Snippet: The SIR and SRIC mRNA vaccines elicit robust humoral immune response. (A) Schematic diagram of SIR and SIRC mRNA vaccine composition, depicting three classes of viral antigen constructs. Three representative SARS-CoV-2 clades, D614G, BA.5, XBB.1.5, four influenza virus strains, A/Wisconsin/67/2022 (H1N1), A/Beijing/32/1992 (H3N2), B/Brisbane/60/2008 (B/Victoria lineage), and B/Florida/4/2006 (B/Yamagata lineage), and RSV A2, were selected for incorporation into a potential 8-valent combination mRNA vaccine formulation. RBD, receptor-binding domain; S, spike protein; HAhr, hemagglutinin head region; HA, hemagglutinin. RBD, receptor-binding domain; S, spike protein; HAhr, hemagglutinin head region; HA, hemagglutinin, Fd, trimer foldon. (B) Representative size distribution of mRNA-LNPs by DLS. (C) PDI of mRNA-LNPs by dynamic light-scattering (DLS). PDI, polymer dispersity index. (D) Encapsulation efficiency of mRNA-LNPs. (E, F, G) HA-, RBD- or pre-F-specific IgG antibody titers at week 6 (2 weeks after the second boost) determined by end-point ELISA. Immunization schedule: Female BALB/c mice (6–8 weeks old, n = 5 per group) were intramuscularly administered 15 μg/100 μL of SIR vaccine (comprising 5 μg of RBD from D614G, BA.5, XBB.1.5, with each contributing 1.7 μg, 5 μg of HAhr mRNA from H1N1 WI22, H3N2 BJ92, B/V BR08, B/Y FL06, with each contributing 1.3 μg, and 5 μg of pre-F mRNA from RSV A2), or 5 μg/100 μL of SARS-CoV-2 vaccine (comprising 5 μg of RBD mRNA from D614G, BA.5, XBB.1.5, with each contributing 1.7 μg), influenza vaccine (comprising 5 μg of HAhr mRNA from H1N1 WI22, H3N2 BJ92, B/V BR08, B/Y FL06, with each contributing 1.3 μg), RSV vaccine (comprising 5 μg of pre-F mRNA from RSV A2). Control groups received 100 μL of PBS. (H) Binding antibody titer profile with values plotted below the bars, showing robust binding activity of the vaccines against a range of viral antigens. (I, J, K) HA-, RBD- or pre-F-specific IgG2a/IgG1 ratio. (L) IgG2a/IgG1 ratio profile with values in the boxs. Green represents a ratio greater than 1, whereas red indicates a ratio less than 1. Blank boxes are marked with an “X.” (M) Neutralizing antibody titers at week 6, expressed as half-maximal inhibitory dilutions (ID 50 ) values against vaccine-matched and mismatched SARS-CoV-2 pseudoviruses. (N, O) Neutralizing antibody titers at week 6 against RSV and four heterologous influenza virus strains. Data are presented as means ± SD. Statistical significance was analyzed by two-way ANOVA with Sidak’s multiple comparisons test or one-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: The human RSV A2 strain (VR-1540, ATCC) and a recombinant derivative, human RSV-A2-mKate2 (pSynkRSVA2 D46F), were generously donated by Dr. Barney S. Graham (VRC, NIH).

Techniques: Vaccines, Construct, Virus, Formulation, Binding Assay, Polymer, Encapsulation, Enzyme-linked Immunosorbent Assay, Control, Activity Assay

Journal: ACS Nano

Article Title: Combination mRNA Vaccine Adjuvanted with CpG Oligodeoxynucleotides Enhances Protection against Respiratory Virus Infection

doi: 10.1021/acsnano.5c14408

Figure Lengend Snippet: SIRC mRNA vaccine protects mice from influenza virus and RSV challenge. Influenza virus and RSV challenges were conducted at week 6 (2 weeks after the second boost) in mice. (A) Longitudinal monitoring of body weight and survival following influenza virus challenge. (B) Representative histopathology H&E images of lung tissue from influenza-challenged mice. Scale bar = 50 μm, 20×. Red arrow, bleeding; Yellow arrow, lymphocyte infiltration; Green arrow, epithelial cells or epithelioid cells. (C) Quantification of RSV viral titers in lung homogenates postchallenge. (D, E) Histopathological scoring of lung tissues based on a 0–4 severity scale for perivasculitis, peribronchiolitis, interstitial pneumonia, and alveolar catarrh, with total pathology scores shown. (F) Representative histopathology H&E images of lung tissue from RSV A2-challenged mice. Scale bar = 50 μm, 20×. Orange arrow, lymphocyte infiltration; Green arrow, epithelial cells or epithelioid cells. Data are presented as means ± standard deviations (SD). Statistical significance was analyzed by two-way ANOVA with Sidak’s multiple comparisons test or one-way ANOVA with Tukey’s multiple comparisons test. Differences in survival rates were assessed using a two-sided log-rank test.

Article Snippet: The human RSV A2 strain (VR-1540, ATCC) and a recombinant derivative, human RSV-A2-mKate2 (pSynkRSVA2 D46F), were generously donated by Dr. Barney S. Graham (VRC, NIH).

Techniques: Virus, Histopathology