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. 2021 Nov 12;9(11):1316. doi: 10.3390/vaccines9111316

Figure 4.

Figure 4

m81 mAb prevents hyperglycemia increase and human IAPP aggregate formation in pancreatic islets, and decreases IL-1β level. (A) Immunization scheme of FVB/Ins2-hIAPP transgenic mice with 100 μg of m81 mAb or control buffer for each injection. Mice received two injections/week. Blood was collected for Ab titer check. (B) Body weight [g] over 13 weeks of m81 mAb immunized (orange square) and control (black circle) mice, with corresponding area under the curve (AUC). (C) 14h-fasting glucose level [mg/dL] of m81 mAb immunized (orange square) and control (black circle) mice, with corresponding area under the curve (AUC). n = at least four mice per group. Statistics performed by unpaired t-test, where the immunized (m81 mAb) group is compared to the control (PBS) group. ** p < 0.01. (D) Glucose Tolerance Test at age of 6 weeks, with corresponding area under the curve (AUC). n = four mice per group. (E) Glucose Tolerance Test at 14 weeks of age, with corresponding area under the curve (AUC) n = four mice per group. Statistics performed by unpaired t-test, where the immunized (m81 mAb) group is compared to the control (Isotype) group. * p < 0.05. (F) Insulin Tolerance Test at age of 6 week, with corresponding area under the curve (AUC). n = four mice per group. (G) Insulin Tolerance Test at age of 14 week, with corresponding area under the curve (AUC). n = four mice per group. (D–G) Statistics performed by unpaired t-test, where the immunized (m81 mAb) group is compared to the control (Isotype) group. ** p < 0.01. (H) Pancreatic islets showing deposited hIAPP (ThioS, green) in control (left panel), m81 mAb immunized (right panel). Nuclei are stained in blue (DAPI). (I) Presence of pro inflammatory cytokine IL-1β (IL-1β, in red), insulin (Ins, in yellow), and nuclei (DAPI, blue) in control (left panel), m81 mAb immunized (right panel) (J) Quantification of insulin (left) and (K) IL-1β positive islets of control (black dot) and m81 mAb immunized (orange square) mice. n = 19 islets. (J–L) Statistics performed by unpaired t-test, where the immunized (m81 mAb) group is compared to the control (PBS) group. ** p < 0.01, *** p < 0.001. m81 mAb clone m81; hIAPP, human amylin; IL-1β, interleukin-1β; ThioS, ThioflavinS, Ins, insulin; DAPI, 4′,6-Diamidine-2′-phenylindole dihydrochloride. Scale bars: 20 μm. (L) IL-1β levels (pg/mL) in the serum of mice (n = 4) at 16 weeks of age determined by ELISA.