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Figure 5 | Alzheimer's Research & Therapy

Figure 5

From: A coimmunization vaccine of Aβ42 ameliorates cognitive deficits without brain inflammation in an Alzheimer’s disease model

Figure 5

Coimmunization suppresses T-cell proliferation by inducing CD25antigen-specific regulatory T cells. Splenocytes were isolated from vaccinated and control mice 7 days after the fifth immunization and used for intracellular staining or for proliferation assays (n = 6). (A) Frequency of FoxP3+ cells in the total CD4+ T cells after the coimmunization compared with other groups by performing fluorescence-activated cell sorting analysis (FACS) analysis. (B) The level of T-cell proliferation was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The splenocytes were restimulated for 3 days in vitro using amyloid-β42 (Aβ42) protein as a specific antigen, bovine serum albumin (BSA) as a nonspecific antigen or anti-CD3 as a positive control. We added anti-CD28 antibody to all T cells as a costimulant. The cell proliferation rate is expressed as the stimulation index (SI). (C) Splenocytes were restimulated for 12 hours in vitro using Aβ42 protein as a specific antigen and blocked with brefeldin A for 8 hours. The lymphocytes were first gated on CD4+ T cells and then used for intracellular staining to detect transforming growth factor β (TGF-β) and interleukin 10 (IL-10), IL-4 and interferon γ (IFN-γ) followed by FACS analysis. A result typical of three independent experiments is shown. All data are presented as mean ± SD. Statistical analysis was performed using parametric one-way analysis of variance, and t-tests were used to compare two groups. *P < 0.05, **P < 0.01 compared with protein-vaccinated model mice. co-Aβ42, APP695 mice immunized with co-amyloid-β42 vaccine; ns, Not significant; pro-Aβ42, APP695 mice immunized with amyloid-β42 protein vaccine; WT, Wild type.

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