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Fig. 1 | Alzheimer's Research & Therapy

Fig. 1

From: Active full-length DNA Aβ42 immunization in 3xTg-AD mice reduces not only amyloid deposition but also tau pathology

Fig. 1

Timeline of the experimental procedures, anti-amyloid-β peptide 1–42 (anti-Aβ42) antibody production upon immunization with DNA Aβ42 trimer and Aβ42 peptide in triple-transgenic Alzheimer’s disease (3xTg-AD) and wild-type mice and cytokine secretion in restimulated splenocyte cultures. a Immunizations, blood draws, and final analyses are shown along the experimental timeline of 20 months. b High levels of anti-Aβ42 antibodies (micrograms per milliliter of plasma) were found in all of the immunized mouse groups following the last immunization (wild-type mice and 3xTg-AD mice). Blue symbols indicate mice that had received DNA Aβ42 trimer immunizations; yellow symbols indicate mice that had received Aβ42 peptide immunizations. Antibody levels of two groups of 20-month-old 3xTg-AD mice are shown as group 1 (G1) and group 2 (G2). Plasma samples had been used in a 1:1000 dilution. Samples were run in triplicates, and the assay was repeated twice. Antibody isotype analyses from DNA Aβ42 trimer-immunized 3xTg-AD mice (c) and Aβ42 peptide-immunized 3xTg-AD mice (d). White bars show levels of anti-Aβ42 antibodies of the immunoglobulin G1 (IgG1) isotype; gray bars show IgG2a antibody levels; hatched bars show IgG2b antibody levels; and black bars show IgM antibody levels. Differences in the amount of IgG1 (Th2) and IgG2a/c (Th1) antibody levels are statistically significant (p = 0.0068). Levels were measured as optical density at 450 nm (OD450). Plasma samples had been used in a 1:500 dilution, analyzed in triplicates, and the assay was repeated twice. e Antibody isotype profile of plasma samples from peptide-immunized mice in a 1:20,000 dilution. Interferon (IFN)-γ (f) and interleukin (IL)-17 (g) enzyme-linked immunospot analysis of splenocytes from 20-month-old 3xTg-AD mice (n = 4/group) and 129/SvJ wild-type mice (n = 4/group) that had received 13 Aβ42 peptide or 13 DNA Aβ42 trimer immunizations, respectively. No IFN-γ- or IL-17-secreting cells were found in DNA Aβ42-immunized mice, whereas high numbers of cells secreting IFN-γ and IL-17 were found in splenocytes from peptide-immunized mice upon Aβ1–42 peptide or Aβ10–26/17–31 peptide mix restimulation in vitro. **, and **** indicate p values of ≤ 0.01 and ≤ 0.001, respectively (unpaired Student's t test)

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