Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: This figure shows serum IgG isotype antibody responses in mice vaccinated with BM5 by intramuscular (IM) N = 7 and Gene Gun (GG), N = 8 immunization. Antigen-specific ELISAs were performed with sera collected 8 wk after the first immunization. A : IgG1 and IgG2a results shown are mean OD + SE for IM and GG mice; B : IgG1 to IgG2a ratios are shown for mice immunized with BM5 by IM and GG routes. Values shown are mean PD +SE. The ratios in the two groups were significantly different (p < 0.05).

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

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Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: This figure shows serum IgG isotype antibody responses in mice vaccinated with BMIF by IM (N = 12) and GG (N = 9). Antigen-specific ELISAs were performed with sera collected 8 wk after immunization. A: IgG1 and IgG2a antibodies shown are mean OD + SE; B: IgG1 to IgG2a ratios are shown for mice immunized with BMIF by IM and GG routes. Values shown are means + SE. The ratios were not significantly different.

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

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Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: This figure shows serum IgG isotype antibody responses in mice vaccinated with BMHSP by IM (N = 12) and GG (N = 9) route. Antigen-specific ELISAs were performed with sera collected 8 wk after immunization. A: IgG1 and IgG2a results shown are mean OD +SE; B: IgG1 to IgG2a ratios are shown for mice immunized with BMHSP by IM and GG route. Values shown are means + SE. The ratios were not significantly different.

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

Techniques:

Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: This figure shows serum IgG isotype antibody responses in mice vaccinated with BM14 by IM (N = 12) and GG (N = 9). Antigen-specific ELISAs were performed with sera collected 8 wk after immunization. A: IgG, IgG1 and IgG2a antibody results shown are mean OD + SE; B: IgG1 to IgG2a antibody ratios are shown for mice immunized with BMHSP by IM and GG routes. Values shown are means + SE. The ratios were not significantly different.

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

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Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: Time course of total IgG antibodies to individual recombinant B. malayi antigens after polyvalent and monovalent DNA vaccination by IM injection. ELISAs were performed with sera collected at different times after immunization. Results shown are mean OD + SE. A: IgG antibody responses to individual antigens after polyvalent DNA vaccination (5 mice). B: IgG antibody responses to individual antigens after monovalent DNA vaccination (7 mice for BM5, 12 mice for each of the other antigens).

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

Techniques: Recombinant, Injection

Journal: ACS infectious diseases

Article Title: Tyrosine-Based Cross-Linking of Peptide Antigens to Generate Nanoclusters with Enhanced Immunogenicity: Demonstration Using the Conserved M2e Peptide of Influenza A.

doi: 10.1021/acsinfecdis.1c00219

Figure Lengend Snippet: Figure 6. In vitro antigenicity analysis of t-M2e-t NCs using ELISA. (A) Absorbance values from an indirect ELISA performed by coating 96-well plates with a fixed amount of either un-cross-linked t-M2e-t peptide, cross-linked t-M2e-t NCs, or a negative control peptide (NCP). Mouse serum collected from mice vaccinated with a gold nanoparticle and M2e formulation (from another study8−10) was used as a source of M2e detection IgG antibody, which was applied at different dilutions. (B) Absorbance values from a sandwich ELISA performed by coating 96-well plates with a fixed amount of purified mouse anti-M2e IgM monoclonal antibody as capture antibody. Dilutions of analyte (either un-cross-linked t-M2e-t peptide, cross-linked t-M2e-t NCs, or a NCP) were then added to the wells. Mouse serum collected from mice vaccinated with a gold nanoparticle and M2e formulation (from another study8−10) was used as a source of primary M2e detection IgG antibody. Cartoons of each ELISA procedure are shown below their respective graphs.

Article Snippet: All peptides were end-terminal acetylated and amidated in order to increase peptide stability against degradation.65−67 Goat anti-mouse IgG (1030−05), goat anti-mouse IgG1 (1070−05), and goat anti-mouse IgG2a (1080−05) secondary antibodies with HRP were obtained from Southern Biotech (AL, USA).

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Indirect ELISA, Negative Control, Formulation, Sandwich ELISA

Journal: ACS infectious diseases

Article Title: Tyrosine-Based Cross-Linking of Peptide Antigens to Generate Nanoclusters with Enhanced Immunogenicity: Demonstration Using the Conserved M2e Peptide of Influenza A.

doi: 10.1021/acsinfecdis.1c00219

Figure Lengend Snippet: Figure 7. Immune response and survival data of mice after immunization with t-M2e-t NCs. Mice were immunized thrice, once each on day 0, 21, and 42. Blood was collected and analyzed for anti-M2e antibodies. Within 1 week of the final serum collection, mice were challenged with 3× LD50 A/California/07/2009 (H1N1) and monitored for 14 days. Two antigens were used (i) 20 μg or 5 μg t-M2e-t UCP (un-cross-linked peptide) with/without 20 μg CpG, and (ii) 20 μg or 5 μg S t-M2e-t NCs (small NCs) with/without 20 μg CpG. (A−C) Anti-M2e IgG, IgG1 and IgG2a titers with t-M2e-t UCP as antigen. (D−F) Anti-M2e IgG, IgG1 and IgG2a titers with t-M2e-t NC as antigen. (G, H) Body weight and survival after virus challenge of t-M2e-t UCP groups. (I, J) Body weight and survival after virus challenge of t-M2e-t NP groups. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: All peptides were end-terminal acetylated and amidated in order to increase peptide stability against degradation.65−67 Goat anti-mouse IgG (1030−05), goat anti-mouse IgG1 (1070−05), and goat anti-mouse IgG2a (1080−05) secondary antibodies with HRP were obtained from Southern Biotech (AL, USA).

Techniques: Virus

Journal: Cells

Article Title: Caspase-1-Dependent Pyroptosis Mediates Adjuvant Activity of Platycodin D as an Adjuvant for Intramuscular Vaccines.

doi: 10.3390/cells11010134

Figure Lengend Snippet: Figure 8. Caspase-1 mediated the adjuvant activity of PD. (A,B) Mice were injected i.m. with z-VAD-FMK (A) or Ac-YVAD-CMK (B) at the dose of 1 µg/g for 1 h before immunization. Sera were collected 2 weeks after the secondary immunization, and serum OVA-specific IgG, IgG1, IgG2a, and IgG2b antibodies were measured by an indirect ELISA. (C–E) Mice were injected i.m. with Ac-YVAD-CMK at the dose of 1 µg/g for 1 h before immunization. Splenocytes were prepared 2 weeks after the secondary immunization. Splenocyte proliferation (C) and NK cell activity (D) were measured by the MTT method. (E) The culture supernatants were measured for the levels of IFN-γ and IL-10 by ELISA. Data were presented as mean ± SD (n = 3). (*) p < 0.05, (**) p < 0.01, (***) p < 0.001, ns not significant.

Article Snippet: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), concanavalin A (Con A), lipopolysaccharide (LPS), acridine orange (AO), ovalbumin (OVA), collagenase II, and rabbit anti-mouse IgG horseradish peroxidase (HRP)-conjugate (#A-9044) were purchased from Sigma-Aldrich, Saint Louis, MO, USA; fetal bovine serum (FBS) was obtained from Gibco, Grand Island, NY, USA; DMEM medium was obtained from Corning, Corning, NY, USA; goat anti-mouse IgG1 (#1070-05), IgG2a (#1080-05), and IgG2b (#1090-05) peroxidase conjugates were acquired from SouthernBiotech, Birmingham, AL, USA; mouse cytokine and chemokine detecting enzyme-linked immunosorbent assay (ELISA) kits were obtained from Boster Biological Technology co., Ltd., Wuhan, China; cyclic 3′,5′-adenosine monophosphate (cAMP) assay kits were acquired from Nanjing Jiancheng Bioengineering Institute, Nanjing, China; Fluo-3 AM was obtained from Dojindo Laboratories, Kumamoto, Japan; bicinchoninic acid (BCA) protein assay kit, reactive oxygen species (ROS) assay kit, enhanced chemiluminescence (ECL) kit, HRP-conjugated goat anti-rabbit (#A0208) and anti-mouse (#A0216) IgG (H+L), and radioimmunoprecipitation assay (RIPA) lysis buffer were acquired from Beyotime Biotech, Nantong, China; and the chromogenic end-point tachypleus amebocyte lysate (CE TAL) was obtained from Xiamen Bioendo Technology Co., Ltd., Xiamen, China.

Techniques: Adjuvant, Activity Assay, Injection, Indirect ELISA, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: CD200R1 and CD200R1L expression is regulated during B cell development in swine and modulates the Ig production in response to the TLR7 ligand imiquimoid

doi: 10.1371/journal.pone.0251187

Figure Lengend Snippet: Characteristics of mAbs used in this study.

Article Snippet: To examine the reactivity of PCT1 and PCT3 mAbs on B cell subpopulations defined according to the expression of CD21 and CD11R1, PBMCs (1 x 10 6 /well) were incubated with anti- porcine CD21 and PCT3 mAbs followed by staining with PE-conjugated goat anti-mouse IgG1 and PE-Cy7-conjugated goat anti-mouse IgG2a polyclonal antibodies (both from Southern Biotech, USA).

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