Journal: Frontiers in Immunology

Article Title: Treatment With CD52 Antibody Protects Neurons in Experimental Autoimmune Encephalomyelitis Mice During the Recovering Phase

doi: 10.3389/fimmu.2021.792465

Figure Lengend Snippet: Treatment with CD52 antibody improves symptoms and pathological changes of C57BL/6J EAE mice. EAE mice established by vaccinating C57BL/6J mice with MOG35-55 were treated with CD52 antibodies or PBS at the peak of disease (~ 16 dpi). Treatments with anti-CD52 significantly attenuated clinical scores of EAE mice [ (A) two-way ANOVA, F (1, 11) = 33.07; n ≥ 5 per group], and increased the body weight, although it was not statistically significant [ (B) two-way ANOVA, F (1, 11) = 2.392; n ≥ 5 per group]. Two weeks after treatments, EAE mice were analyzed for axonal degeneration and myelin loss. Treatments with CD52 antibodies significantly reduced APP-positive spheroids (in brown) [ (C, D) t test, t (7) = 4.485; n ≥ 4 per group], and markedly increased the coverage of MOG-positive myelin (in green) [ (E, F) t test, t (11) = 2.978; n ≥ 5 per group] in the white matter of anterior and lateral columns at the lumber spinal cord (as shown in E with the frame), compared with PBS-treated EAE mice. The presented images are from EAE mice 14 days post treatments. Interestingly, the number of APP-positive spheroids was negatively correlated with the area of MOG-positive myelin [ (G) Pearson correlation test; n = 9]. EAE mice were also analyzed within 4 days after treatments. Anti-CD52 treatment immediately reduced the clinical scores of EAE mice [ (H) two-way ANOVA, F (1, 15) = 17.24; n ≥ 8 per group], and increased the body weight, although not statistically significant [ (I) two-way ANOVA, F (1, 15) = 1.515; n ≥ 8 per group]. Histological analysis showed that treatments with anti-CD52 antibodies significantly decreased the number of APP-positive spheroids [ (J) t test, t (12) = 2.339; n = 7 per group] but did not change the coverage of MOG-positive myelin in the white matter of lumber spinal cord [ (K) t test, t (16) = 1.581; n ≥ 8 per group].

Article Snippet: Eight-week-old C57BL/6J mice, and BDNF-ablated and wild-type littermate mice were s.c. immunized with 100μg mouse MOG35-55 (MEVGWYRSPFSRVVHLYRNGK, EZBiolab, Carmel, USA) in a complete Freund’s adjuvant (BD Biosciences, Heidelberg, Germany) supplemented with 100μg Mycobacterium tuberculosis (H37Ra) (BD Biosciences), followed by i.v. injection of Pertussis toxin (PTX, Enzo Life Sciences GmbH, Lörrach, Germany) 0 and 2 days after MOG immunization (d.p.i).

Techniques:

Journal: Frontiers in Immunology

Article Title: Treatment With CD52 Antibody Protects Neurons in Experimental Autoimmune Encephalomyelitis Mice During the Recovering Phase

doi: 10.3389/fimmu.2021.792465

Figure Lengend Snippet: Treatment with CD52 antibody improves symptoms and pathology in both neuronal BDNF-deficient and wildtype EAE mice. Eight-week-old C57BL/6 littermate mice with (ko) and without (wt) BDNF deficiency in neurons were immunized with MOG35-55 in a complete Freund’s adjuvant. The reduction of BDNF expression in neurons of spinal cord was detected by immunological staining of BDNF and NeuN [ (A) BDNF-expressing neurons are marked with arrow heads] and quantitated by real-time PCR measurement of bdnf gene transcripts in the cervical spinal cord [ (B) t test; t (6) = 4.444; n ≥ 3 per group], and Western blot evaluation of pro-BDNF in the brain homogenate (C, D) t test; t (9) = 3.483; n ≥ 4 per group). At the peak of disease (around 16 dpi), CD52 antibodies were subcutaneously administered for 5 days. PBS was injected as a control. Clinical symptoms were monitored for around 2 weeks after anti-CD52 treatments. The clinical scores were not significantly different between BDNF wt and ko mice after injection with PBS [ (E) two-way ANOVA followed by Bonferroni post-hoc test, p = 0.718]; whereas the clinical scores of both BDNF wt and ko mice were significantly reduced by treatments with CD52 antibodies as compared with EAE mice receiving PBS injection [ (E) two-way ANOVA followed by Bonferroni post-hoc test; F (3, 15) = 11.02; n ≥ 3 per group]. Interestingly, the anti-CD52 treatments-induced recovering of BDNF-wt EAE mice was significantly better than of BDNF-ko EAE mice [ (E) two-way ANOVA followed by Bonferroni post-hoc test, p = 0.001]. Myelination in BDNF-ko EAE mice were further evaluated by immunofluorescent staining of MOG (F) . Treatments with CD52 antibodies compared with PBS treatments significantly increased the coverage of MOG-positive myelin (in green) in the white matter of anterior horn of the lumber spinal cord (as shown with the frame) (G) one-way ANOVA followed by Bonferroni post-hoc test; F (3, 19) = 7.858; n ≥ 3 per group).

Article Snippet: Eight-week-old C57BL/6J mice, and BDNF-ablated and wild-type littermate mice were s.c. immunized with 100μg mouse MOG35-55 (MEVGWYRSPFSRVVHLYRNGK, EZBiolab, Carmel, USA) in a complete Freund’s adjuvant (BD Biosciences, Heidelberg, Germany) supplemented with 100μg Mycobacterium tuberculosis (H37Ra) (BD Biosciences), followed by i.v. injection of Pertussis toxin (PTX, Enzo Life Sciences GmbH, Lörrach, Germany) 0 and 2 days after MOG immunization (d.p.i).

Techniques: Expressing, Staining, Real-time Polymerase Chain Reaction, Western Blot, Injection

Journal: Frontiers in Immunology

Article Title: Treatment With CD52 Antibody Protects Neurons in Experimental Autoimmune Encephalomyelitis Mice During the Recovering Phase

doi: 10.3389/fimmu.2021.792465

Figure Lengend Snippet: Deficiency of neuronal BDNF attenuates CD52 antibody-induced reduction of inflammatory infiltrates in EAE mice. Eight-week-old C57BL/6 littermate mice with (ko) and without (wt) BDNF deficiency in neurons were immunized with MOG35-55 in a complete Freund’s adjuvant. EAE mice were administered subcutaneously with CD52 antibodies or PBS for 5 days at the peak of disease. Two weeks after treatments, the lumber segment of spinal cord was analyzed with immunohistochemistry for the infiltration of T cells and microglia/macrophages (A, D) immune reactive cells are labelled in brown). Treatments of CD52 antibodies significantly reduced the number of CD3-positive cells (B) one-way ANOVA followed by Bonferroni post-hoc test; F (3, 15) = 42.81; n ≥ 3 per group) and tended to decrease Iba1-positive cells (E) one-way ANOVA followed by Bonferroni post-hoc test; F (3, 16) = 21.20; n ≥ 3 per group) in BDNF-ko EAE mice. Interestingly, deficiency of neuronal BDNF significantly decreased the number of both CD3 and Iba1-positive cells in the lumber spinal cord of PBS-treated EAE mice (B, E) . BDNF deficiency also significantly attenuated CD52 antibody-induced reduction of CD3- and Iba1-positive cells relative to PBS-treated EAE mice (C, F) t test; t (7) = 6.014 and t (8) = 4.496 for CD3 and Iba1-positive cells, respectively; n ≥ 4 per group).

Article Snippet: Eight-week-old C57BL/6J mice, and BDNF-ablated and wild-type littermate mice were s.c. immunized with 100μg mouse MOG35-55 (MEVGWYRSPFSRVVHLYRNGK, EZBiolab, Carmel, USA) in a complete Freund’s adjuvant (BD Biosciences, Heidelberg, Germany) supplemented with 100μg Mycobacterium tuberculosis (H37Ra) (BD Biosciences), followed by i.v. injection of Pertussis toxin (PTX, Enzo Life Sciences GmbH, Lörrach, Germany) 0 and 2 days after MOG immunization (d.p.i).

Techniques: Immunohistochemistry