Methods to monitor monocytes-mediated amyloid-beta uptake and phagocytosis in the context of adjuvanted immunotherapies

J Immunol Methods. 2015 Sep:424:64-79. doi: 10.1016/j.jim.2015.05.002. Epub 2015 May 19.

Abstract

Antibody-mediated capture of amyloid-beta (Aβ) in peripheral blood was identified as an attractive strategy to eliminate cerebral toxic amyloid in Alzheimer's disease (AD) patients and murine models. Alternatively, defective capacity of peripheral monocytes to engulf Aβ was reported in individuals with AD. In this report, we developed different approaches to investigate cellular uptake and phagocytosis of Aβ, and to examine how two immunological devices--an immunostimulatory Adjuvant System and different amyloid specific antibodies--may affect these biological events. Between one and thirteen months of age, APPswe X PS1.M146V (TASTPM) AD model mice had decreasing concentrations of Aβ in their plasma. In contrast, the proportion of blood monocytes containing Aβ tended to increase with age. Importantly, the TLR-agonist containing Adjuvant System AS01B primed monocytes to promote de novo Aβ uptake capacity, particularly in the presence of anti-Aβ antibodies. Biochemical experiments demonstrated that cells achieved Aβ uptake and internalization followed by Aβ degradation via mechanisms that required effective actin polymerization and proteolytic enzymes such as insulin-degrading enzyme. We further demonstrated that both Aβ-specific monoclonal antibodies and plasma from Aβ-immunized mice enhanced the phagocytosis of 1 μm Aβ-coated particles. Together, our data highlight a new biomarker testing to follow amyloid clearance within the blood and a mechanism of Aβ uptake by peripheral monocytes in the context of active or passive immunization, and emphasize on novel approaches to investigate this phenomenon.

Keywords: Adjuvant; Alzheimer's disease; Amyloid-beta; Aβ uptake; Monocyte; Phagocytosis; Vaccine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Adjuvants, Immunologic
  • Alzheimer Disease / immunology
  • Alzheimer Disease / metabolism
  • Alzheimer Disease / therapy
  • Amyloid beta-Peptides / blood
  • Amyloid beta-Peptides / immunology
  • Amyloid beta-Peptides / metabolism*
  • Animals
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / metabolism
  • Brain / immunology
  • Brain / metabolism
  • Brain / pathology
  • Cell Line
  • Disease Models, Animal
  • Drug Combinations
  • Immunophenotyping
  • Immunotherapy
  • Lipid A / administration & dosage
  • Lipid A / analogs & derivatives
  • Lipid A / immunology
  • Mice
  • Mice, Transgenic
  • Monocytes / immunology*
  • Monocytes / metabolism*
  • Phagocytosis / immunology*
  • Protein Multimerization
  • Proteolysis
  • Saponins / administration & dosage
  • Saponins / immunology
  • Vaccination

Substances

  • Actins
  • Adjuvants, Immunologic
  • Amyloid beta-Peptides
  • Antibodies, Monoclonal
  • Drug Combinations
  • Lipid A
  • Saponins
  • adjuvant system 01