Journal: Frontiers in Immunology

Article Title: Females with obesity exhibit greater influenza vaccine-induced immunity and protection than males in a mouse model

doi: 10.3389/fimmu.2025.1699275

Figure Lengend Snippet: Vaccine-induced antibody responses in males and females, with or without obesity. Male and female C57BL/6J mice, with or without obesity were vaccinated twice with the 2009 pandemic H1N1 influenza A virus (IAV) vaccine at a 3-week interval. At 35 days post-vaccination (dpv), blood samples were collected, and antibodies were measured in plasma. (A) IgG, (B) IgG1, (C) IgG2c, (D) IgG1 to IgG2c ratio, and (E) virus-neutralizing antibody (nAb) titers are compared (dashed line – limit of detection; n=13-15/group). Subjects that had an antibody titer below the limit of detection are shown in percent in each figure. Statistical comparisons were made using two-way analysis of variance (ANOVA) (A-E) followed by Tukey’s multiple comparisons. An asterisk (*) indicates a significant difference (p<0.05) and a hash ( # ) represents a trend (0.05≤p≤0.1). NO-M, non-obese males; NO-F, non-obese females; O-M, males with obesity; O-F, females with obesity.

Article Snippet: Secondary antibodies used were IgG (#31430, Invitrogen, CA, USA), IgG1 (#PA1-74421, Invitrogen, CA, USA), and IgG2c (#56970, Cell Signaling Technologies, MA, USA), followed by the tetramethylbenzidine (TMB) substrate reagent (#555214, BD Biosciences, CA, USA) addition (50μL/well).

Techniques: Virus, Clinical Proteomics

Journal: Frontiers in Immunology

Article Title: Females with obesity exhibit greater influenza vaccine-induced immunity and protection than males in a mouse model

doi: 10.3389/fimmu.2025.1699275

Figure Lengend Snippet: Comparisons of immunity and protection between non-responder females (i.e., NR-F) and responder females with obesity (i.e., R/O-F). Following 12 weeks on HFD, nearly 2/3 rd of females developed obesity, and the remainder were non-responders. (A) Baseline body mass; (B) body mass over 12 weeks on HFD; (C) week 12 body mass; (D) body mass index (BMI); (E) glucose tolerance test (GTT) time course (0–120 min); and (F) GTT area under the curve (AUC) are shown (n=9-19/group). After prime-boost vaccination with the inactivated 2009 H1N1 influenza vaccine, at 35 days post-vaccination (dpv), (G) IgG, (H) IgG1, (I) IgG2c, (J) IgG1/IgG2c, and (K) virus-neutralizing antibody (nAb) titers were measured (dashed line – limit of detection, n=9-15/group). Following the challenge with the drift variant of H1N1 virus, (L) absolute (g) and (M) relative (%) body mass change through 21 days post-challenge (dpc); (N) lung viral titers at 3 dpc; and (O) log 10 -transformed cytokines/chemokines concentrations at 3 dpc were compared (n=3-5/group). Statistical comparisons were made using the (A, C, D, F, G-K, N) Mann-Whitney test, (B, E, L, M) two-way repeated measures analysis of variance (ANOVA) followed by Tukey’s multiple comparisons, and (O) , unpaired T-test followed by Holm-Sidak test for multiple comparisons. An asterisk (*) indicates a significant difference (p<0.05) and a hash ( # ) represents a trend (0.05≤p≤0.1).

Article Snippet: Secondary antibodies used were IgG (#31430, Invitrogen, CA, USA), IgG1 (#PA1-74421, Invitrogen, CA, USA), and IgG2c (#56970, Cell Signaling Technologies, MA, USA), followed by the tetramethylbenzidine (TMB) substrate reagent (#555214, BD Biosciences, CA, USA) addition (50μL/well).

Techniques: Virus, Variant Assay, Transformation Assay, MANN-WHITNEY

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Neutral Polymer Micelle Carriers with pH-Responsive, Endosome-Releasing Activity Modulate Antigen Trafficking to Enhance CD8 T-Cell Responses

doi: 10.1016/j.jconrel.2014.03.041

Figure Lengend Snippet: Splenocytes were isolated on day 29 from mice immunized twice, 3 weeks apart, with free ova, physical mixture, and pH-responsive conjugate. a) Cells were plated and tested for IFN-γ in vitro recall responses by incubating with CD8+ T cell epitope ova257-264 (SINFEKL). CD8+ T cells were identified by ICS based on CD8 expression, and the percentage of these cells expressing IFN-γ determined. Means ± SEM (n = 7–15) shown are pooled from three independent experiments. Representative flow cytometry dot plots of CD8+ IFN-γ+ T cells from individual mice. b) Ova-specific IgG1 (black bars) and IgG2c (white bars) antibody titres were measured on day 28 in sera from immunized mice. Data shown are pooled from four independent experiments and represent the mean of the reciprocal dilution ± SEM (n = 7–15). One-way ANOVA followed by Tukey’s Multiple Comparison Test was used for all statistical analyses. *p < 0.05.

Article Snippet: Post incubation plates were washed with PBST and incubated with biotin-conjugated anti-mouse antibodies to IgG1 (BD Pharmingen) or IgG2c (Bethyl Laboratories) at 0.005 μg/mL in 0.1% BSA/PBST for 1 h at RT.

Techniques: Isolation, In Vitro, Expressing, Flow Cytometry

Journal: Heliyon

Article Title: Poly(I:C) and R848 ligands show better adjuvanticity to induce B and T cell responses against the antigen(s)

doi: 10.1016/j.heliyon.2024.e26887

Figure Lengend Snippet: The antibody response following the repeated immunization with OVA when injected with adjuvants. OVA specific IgG response was measured by the indirect ELISA after each immunization (n=3 mice/group) in, (A) immunization with OVA (B) OP (Ovalbumin protein and poly(I:C) (C) OR (Ovalbumin protein and R848). The endpoint titer was measured in (D) OVA, (E) OP and, (F) OR group. Similarly, OVA specific IgG1 and, IgG2c response was measured in (G & J) OVA, (H & K) OP & (I & L) OR group of immunization, respectively. The bar represents mean SD for each dose administered in each group at different dilutions. The P values were determined by comparing means using two-way analysis of variance (ANOVA) test followed by Tukey's multiple comparison test for IgG, IgG1 and IgG2c (between 1 st dose and 3 rd dose). ****P<0.0001, ***P=0.0002, **P=0.021, *P=0.0332, ns=0.1234. For endpoint titer, p values were determined by comparing means using one-way ANOVA test followed by Tukey's multiple comparison test.

Article Snippet: Bovine serum albumin (BSA) fraction-V, cell culture tested (Himedia: TC194-100G), goat anti-mouse IgG (H+L) Secondary Ab), HRP (In-vitrogen 62-6520), goat anti-mouse IgG1 Secondary Ab, HRP (In-vitrogen PA1-74421), goat anti-mouse IgG2c, Fc gamma specific Ab, HRP (CST 56970), 3,3′,5,5′-Tetramethylbenzidine (TMB) Liquid Substrate System for ELISA (Sigma T0440-100ML), carbonate-bicarbonate buffer (Sigma C3041-50CAP), eBioscience TM 10X RBC lysis buffer (Multi-species) (Invitrogen 00-4300-54), OVA-Albumin from chicken egg white, lyophilized powder ≥98% electrophoresis (Sigma A5503-5G), 10X PBS (Himedia ML023-500ML), 10X PBS (Endotoxin Free) (Himedia ML164-100ML), RPMI-1640 (Himedia AL199A-500ML), antibiotic antimycotic solution 100x liquid (Himedia A002-20ML), L-Glutamine 200mM solution (Himedia TCL012-20ML), HEPES-1M Solution Cell culture tested (Himedia TCL021), phosphate buffer saline with 0.05% tween 20, pH 7.4 (Sigma P3563-10PAK), ELISA Plate (Immunoplate Strip Single Well-Genetix-38296 or 96 well, high binding, detachable (HiMedia-EP2-5X10NO)), Poly I:C (LMW-25mg) (InVivogen-tlrl-picw), R848 (Sigma SMLL0196-10MG), BD Cytofix/Cytoperm TM Plus Fixaion/Permeabilization solution kit with BD GolgiPlug TM (BD 555028).

Techniques: Injection, Indirect ELISA, Comparison

Journal: Heliyon

Article Title: Poly(I:C) and R848 ligands show better adjuvanticity to induce B and T cell responses against the antigen(s)

doi: 10.1016/j.heliyon.2024.e26887

Figure Lengend Snippet: Antibody titer and antibody avidity index after the repeated immunization with OVA protein supplied with the combination of adjuvants . OVA specific (A) IgG, (B) IgG1, (C) IgG2c, (D) endpoint titer and, (E & F) antibody avidity index (AI) was measured by the indirect ELISA after each immunization (n=3 mice/group) in immunization with OPR (Ovalbumin protein+ poly(I:C)+R848). The bar represents mean SD for each dose administered in each group at different dilutions. The P values were determined by comparing means using two-way analysis of variance (ANOVA) test followed by Tukey's multiple comparison test for IgG, IgG1 and IgG2c (between 1 st dose and 3 rd dose). ****P<0.0001, ***P=0.0002, **P=0.021, *P=0.0332, ns=0.1234. For endpoint titer, p values were determined by comparing means using one-way ANOVA test followed by Tukey's multiple comparison test.

Article Snippet: Bovine serum albumin (BSA) fraction-V, cell culture tested (Himedia: TC194-100G), goat anti-mouse IgG (H+L) Secondary Ab), HRP (In-vitrogen 62-6520), goat anti-mouse IgG1 Secondary Ab, HRP (In-vitrogen PA1-74421), goat anti-mouse IgG2c, Fc gamma specific Ab, HRP (CST 56970), 3,3′,5,5′-Tetramethylbenzidine (TMB) Liquid Substrate System for ELISA (Sigma T0440-100ML), carbonate-bicarbonate buffer (Sigma C3041-50CAP), eBioscience TM 10X RBC lysis buffer (Multi-species) (Invitrogen 00-4300-54), OVA-Albumin from chicken egg white, lyophilized powder ≥98% electrophoresis (Sigma A5503-5G), 10X PBS (Himedia ML023-500ML), 10X PBS (Endotoxin Free) (Himedia ML164-100ML), RPMI-1640 (Himedia AL199A-500ML), antibiotic antimycotic solution 100x liquid (Himedia A002-20ML), L-Glutamine 200mM solution (Himedia TCL012-20ML), HEPES-1M Solution Cell culture tested (Himedia TCL021), phosphate buffer saline with 0.05% tween 20, pH 7.4 (Sigma P3563-10PAK), ELISA Plate (Immunoplate Strip Single Well-Genetix-38296 or 96 well, high binding, detachable (HiMedia-EP2-5X10NO)), Poly I:C (LMW-25mg) (InVivogen-tlrl-picw), R848 (Sigma SMLL0196-10MG), BD Cytofix/Cytoperm TM Plus Fixaion/Permeabilization solution kit with BD GolgiPlug TM (BD 555028).

Techniques: Indirect ELISA, Comparison

Journal: The Journal of Immunology Author Choice

Article Title: The Regulation of Nucleic Acid Vaccine Responses by the Microbiome

doi: 10.4049/jimmunol.2300196

Figure Lengend Snippet: Details of Ab reagents used in these experiments

Article Snippet: OVA-specific IgG2c , None , 3E3A9 , Chondrex , 7109 , .

Techniques: Conjugation Assay, Concentration Assay

Journal: The Journal of Immunology Author Choice

Article Title: The Regulation of Nucleic Acid Vaccine Responses by the Microbiome

doi: 10.4049/jimmunol.2300196

Figure Lengend Snippet: Humoral and cellular responses to OVA immunization in GF and SPF mice. (A) OVA-specific IgG1, IgG2c, and IgG2b titers in serum and OVA-stimulated splenocytes. (B) IFN-γ, (C) IL-4, and (D) IL-10 responses from GF or SPF mice either unimmunized (naive) (n = 3) or immunized with with OVA in PBS (n = 17–19), OVA with AS01 (AS01) (n = 6–8), OVA with Alum (Alum) (n = 7–8), or OVA with AddaVax (Addavax) (n = 3–4). ICS flow cytometry was pregated on live, single, CD3+ CD4+ cells. Samples falling below the level of cytokine detection by ELISA were set at half the level of cytokine detection, which is the beginning of the y-axis, for visualization. Each data point represents an individual mouse from one experiment. Vaccine-matched GF and SPF groups were compared for statistical significance by Mann–Whitney U test, *p < 0.05, **p < 0.01.

Article Snippet: OVA-specific IgG2c , None , 3E3A9 , Chondrex , 7109 , .

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY