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Comparing the efficacy and neuroinflammatory potential of three anti-abeta antibodies.

Fuller JP, Stavenhagen JB, Christensen S, Kartberg F, Glennie MJ, Teeling JL - Acta Neuropathol. (2015)

Bottom Line: In this study, we directly compared 3 clinical candidates in the same pre-clinical model, with the same effector function, for their ability to clear plaques and induce inflammation in the brain.This translated into 3D6 effectively clearing plaques and reducing the levels of insoluble Aβ, whilst chGantenerumab and mC2 did not. 3D6 caused a significant increase in the levels of pro-inflammatory cytokines IL-1β and TNFα, and an associated increase in microglial expression of CD11B and CD68. chGantenerumab increased pro-inflammatory cytokines and microglial activation, but minimal changes in CD68, as an indicator of phagocytosis.Injection of mC2 did not cause any significant inflammatory changes.

View Article: PubMed Central - PubMed

Affiliation: Centre for Biological Sciences, University of Southampton, Southampton, UK. jpf1g11@soton.ac.uk.

ABSTRACT
Immunotherapy is a promising strategy for the treatment of Alzheimer's disease (AD). Antibodies directed against Amyloid Beta (Aβ) are able to successfully clear plaques and reverse cognitive deficits in mouse models. Excitement towards this approach has been tempered by high profile failures in the clinic, one key issue has been the development of inflammatory side effects in the brain (ARIAs). New antibodies are entering the clinic for Alzheimer's disease; therefore, it is important to learn all we can from the current generation. In this study, we directly compared 3 clinical candidates in the same pre-clinical model, with the same effector function, for their ability to clear plaques and induce inflammation in the brain. We produced murine versions of the antibodies: Bapineuzumab (3D6), Crenezumab (mC2) and Gantenerumab (chGantenerumab) with an IgG2a constant region. 18-month transgenic APP mice (Tg2576) were injected bilaterally into the hippocampus with 2 µg of each antibody or control. After 7 days, the mice tissue was analysed for clearance of plaques and neuroinflammation by histology and biochemical analysis. 3D6 was the best binder to plaques and in vitro, whilst mC2 bound the least strongly. This translated into 3D6 effectively clearing plaques and reducing the levels of insoluble Aβ, whilst chGantenerumab and mC2 did not. 3D6 caused a significant increase in the levels of pro-inflammatory cytokines IL-1β and TNFα, and an associated increase in microglial expression of CD11B and CD68. chGantenerumab increased pro-inflammatory cytokines and microglial activation, but minimal changes in CD68, as an indicator of phagocytosis. Injection of mC2 did not cause any significant inflammatory changes. Our results demonstrate that the ability of an antibody to clear plaques and induce inflammation is dependent on the epitope and affinity of the antibody.

No MeSH data available.


Related in: MedlinePlus

Cytokine levels following intracranial injection of anti-Aβ mAbs. Cytokine levels were measured in homogenate made from hippocampal punches taken from Tg2576 mice injected with antibodies. Peptide levels were measured by multiplex ELISA and normalised to total protein concentration. a–f Cytokine levels in hippocampal punches expressed as pg.mg total protein. Injection of 3D6 leads to significantly increased levels of pro-inflammatory cytokines TNFα and IL-1β compared to IgG2a control (ap = 0.0042, bp = 0.0262). Data were analysed by one-way ANOVA and Tukey post hoc test (n = 6/7), and presented as mean ± SD
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Fig5: Cytokine levels following intracranial injection of anti-Aβ mAbs. Cytokine levels were measured in homogenate made from hippocampal punches taken from Tg2576 mice injected with antibodies. Peptide levels were measured by multiplex ELISA and normalised to total protein concentration. a–f Cytokine levels in hippocampal punches expressed as pg.mg total protein. Injection of 3D6 leads to significantly increased levels of pro-inflammatory cytokines TNFα and IL-1β compared to IgG2a control (ap = 0.0042, bp = 0.0262). Data were analysed by one-way ANOVA and Tukey post hoc test (n = 6/7), and presented as mean ± SD

Mentions: We showed that in vitro, all three Aβ antibodies engage FcγRs resulting in macrophage activation and TNFα secretion. Therefore, we next investigated the neuroinflammatory potential in vivo. Injection of 3D6 leads to significantly increased levels of pro-inflammatory cytokines TNFα and IL-1β compared to IgG2a control (Fig. 5a, b, p = 0.0042, p = 0.0262). Increased levels of KC/GRO in 3D6 injected animals were observed, although this did not reach significance (Fig. 5d, p = 0.10). Injection of chGantenerumab also results in increased neuroinflammation, while mC2 did not affect any cytokine levels measured. These observations suggest that high affinity IgG2a anti-Aβ antibodies reduce Aβ load but this is associated with increased neuroinflammation.Fig. 5


Comparing the efficacy and neuroinflammatory potential of three anti-abeta antibodies.

Fuller JP, Stavenhagen JB, Christensen S, Kartberg F, Glennie MJ, Teeling JL - Acta Neuropathol. (2015)

Cytokine levels following intracranial injection of anti-Aβ mAbs. Cytokine levels were measured in homogenate made from hippocampal punches taken from Tg2576 mice injected with antibodies. Peptide levels were measured by multiplex ELISA and normalised to total protein concentration. a–f Cytokine levels in hippocampal punches expressed as pg.mg total protein. Injection of 3D6 leads to significantly increased levels of pro-inflammatory cytokines TNFα and IL-1β compared to IgG2a control (ap = 0.0042, bp = 0.0262). Data were analysed by one-way ANOVA and Tukey post hoc test (n = 6/7), and presented as mean ± SD
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4612324&req=5

Fig5: Cytokine levels following intracranial injection of anti-Aβ mAbs. Cytokine levels were measured in homogenate made from hippocampal punches taken from Tg2576 mice injected with antibodies. Peptide levels were measured by multiplex ELISA and normalised to total protein concentration. a–f Cytokine levels in hippocampal punches expressed as pg.mg total protein. Injection of 3D6 leads to significantly increased levels of pro-inflammatory cytokines TNFα and IL-1β compared to IgG2a control (ap = 0.0042, bp = 0.0262). Data were analysed by one-way ANOVA and Tukey post hoc test (n = 6/7), and presented as mean ± SD
Mentions: We showed that in vitro, all three Aβ antibodies engage FcγRs resulting in macrophage activation and TNFα secretion. Therefore, we next investigated the neuroinflammatory potential in vivo. Injection of 3D6 leads to significantly increased levels of pro-inflammatory cytokines TNFα and IL-1β compared to IgG2a control (Fig. 5a, b, p = 0.0042, p = 0.0262). Increased levels of KC/GRO in 3D6 injected animals were observed, although this did not reach significance (Fig. 5d, p = 0.10). Injection of chGantenerumab also results in increased neuroinflammation, while mC2 did not affect any cytokine levels measured. These observations suggest that high affinity IgG2a anti-Aβ antibodies reduce Aβ load but this is associated with increased neuroinflammation.Fig. 5

Bottom Line: In this study, we directly compared 3 clinical candidates in the same pre-clinical model, with the same effector function, for their ability to clear plaques and induce inflammation in the brain.This translated into 3D6 effectively clearing plaques and reducing the levels of insoluble Aβ, whilst chGantenerumab and mC2 did not. 3D6 caused a significant increase in the levels of pro-inflammatory cytokines IL-1β and TNFα, and an associated increase in microglial expression of CD11B and CD68. chGantenerumab increased pro-inflammatory cytokines and microglial activation, but minimal changes in CD68, as an indicator of phagocytosis.Injection of mC2 did not cause any significant inflammatory changes.

View Article: PubMed Central - PubMed

Affiliation: Centre for Biological Sciences, University of Southampton, Southampton, UK. jpf1g11@soton.ac.uk.

ABSTRACT
Immunotherapy is a promising strategy for the treatment of Alzheimer's disease (AD). Antibodies directed against Amyloid Beta (Aβ) are able to successfully clear plaques and reverse cognitive deficits in mouse models. Excitement towards this approach has been tempered by high profile failures in the clinic, one key issue has been the development of inflammatory side effects in the brain (ARIAs). New antibodies are entering the clinic for Alzheimer's disease; therefore, it is important to learn all we can from the current generation. In this study, we directly compared 3 clinical candidates in the same pre-clinical model, with the same effector function, for their ability to clear plaques and induce inflammation in the brain. We produced murine versions of the antibodies: Bapineuzumab (3D6), Crenezumab (mC2) and Gantenerumab (chGantenerumab) with an IgG2a constant region. 18-month transgenic APP mice (Tg2576) were injected bilaterally into the hippocampus with 2 µg of each antibody or control. After 7 days, the mice tissue was analysed for clearance of plaques and neuroinflammation by histology and biochemical analysis. 3D6 was the best binder to plaques and in vitro, whilst mC2 bound the least strongly. This translated into 3D6 effectively clearing plaques and reducing the levels of insoluble Aβ, whilst chGantenerumab and mC2 did not. 3D6 caused a significant increase in the levels of pro-inflammatory cytokines IL-1β and TNFα, and an associated increase in microglial expression of CD11B and CD68. chGantenerumab increased pro-inflammatory cytokines and microglial activation, but minimal changes in CD68, as an indicator of phagocytosis. Injection of mC2 did not cause any significant inflammatory changes. Our results demonstrate that the ability of an antibody to clear plaques and induce inflammation is dependent on the epitope and affinity of the antibody.

No MeSH data available.


Related in: MedlinePlus