Fig. 6: Monoclonal ASM antibody (23A12C3) inhibits pathogenic Th17 cell differentiation and protects neuropathological features in APP/PS1 mice.
From: Immunotherapy targeting plasma ASM is protective in a mouse model of Alzheimer’s disease
a Effect of various concentrations of 23A12C3 on the direct inhibition of ASM activity (n = 3 independent experiments). b Binding affinity (KD) of 23A12C3 with recombinant human ASM by ELISA (n = 6 independent experiments). c Representative binding sensorgrams depicting the interaction of 23A12C3 with recombinant human ASM assessed by SPR detection. d ASM activity (n = 6/group) and ceramide concentration (n = 4/group) in membrane of CD4+ T cells treated with Smpd1ox/ox serum (from 3-month-old Tie2-cre; Smpd1ox/ox mice) and IgG isotype control or 23A12C3. e Representative flow cytometry plot and graph displaying the calculated percentage and mean fluorescent intensity of Th17 cells (CD4+ IL17A+) differentiated from CD4+ T cell in each group (Control, n = 5; Smpd1ox/ox serum/con IgG ab, n = 10; Smpd1ox/ox serum/23A12C3, n = 8). f Western blotting for p-Stat3, p-JNK, p-AKT, p-mTOR in Th17 cells differentiated from CD4+ T cell in each group (n = 4/group). g ASM activity in plasma (n = 5/group). h Percentage of Th17 cells in the spleen, blood, and brain by flow cytometry (n = 5/group). i Quantification of microglia (Iba-1) and astrocyte (GFAP) (n = 5/group). j mRNA levels of inflammatory markers in the cortex and hippocampus (n = 4/group). k Representative immunofluorescence images and quantification of ThioS (Aβ plaques) in cortex and hippocampus. Scale bars, 50 μm. (n = 5/group). l Quantification of 6E10 (n = 5/group). m The results of Morris water maze test (n = 10/group). n, o At probe trial day 11, time spent in target platform (n) and the number of times each animal entered the small target zone (o) were analyzed (n = 10/group). p The results of fear-conditioning test (n = 10/group). d–j, m–p One-way analysis of variance, Tukey’s post hoc test. k, l, n Two-tailed student’s t test. All error bars indicate s.e.m. All in vivo data analysis was done at 9-mo-old mice. Source data are provided as a Source Data file.
