ABSTRACT
Antibody-mediated capture of amyloid-beta (Aß) 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 Aß was reported in individuals with AD. In this report, we developed different approaches to investigate cellular uptake and phagocytosis of Aß, 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 Aß in their plasma. In contrast, the proportion of blood monocytes containing Aß tended to increase with age. Importantly, the TLR-agonist containing Adjuvant System AS01B primed monocytes to promote de novo Aß uptake capacity, particularly in the presence of anti-Aß antibodies. Biochemical experiments demonstrated that cells achieved Aß uptake and internalization followed by Aß degradation via mechanisms that required effective actin polymerization and proteolytic enzymes such as insulin-degrading enzyme. We further demonstrated that both Aß-specific monoclonal antibodies and plasma from Aß-immunized mice enhanced the phagocytosis of 1 µm Aß-coated particles. Together, our data highlight a new biomarker testing to follow amyloid clearance within the blood and a mechanism of Aß uptake by peripheral monocytes in the context of active or passive immunization, and emphasize on novel approaches to investigate this phenomenon.
Subject(s)
Amyloid beta-Peptides/metabolism , Monocytes/immunology , Monocytes/metabolism , Phagocytosis/immunology , Actins/metabolism , Adjuvants, Immunologic , Alzheimer Disease/immunology , Alzheimer Disease/metabolism , Alzheimer Disease/therapy , Amyloid beta-Peptides/blood , Amyloid beta-Peptides/immunology , 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 , Protein Multimerization , Proteolysis , Saponins/administration & dosage , Saponins/immunology , VaccinationABSTRACT
Alzheimer's disease (AD) is the most common cause of dementia worldwide. The pathogenesis of this neurodegenerative disease, currently without curative treatment, is associated with the accumulation of amyloid ß (Aß) in brain parenchyma and cerebral vasculature. AD patients are unable to clear this toxic peptide, leading to Aß accumulation in their brains and, presumably, the pathology associated with this devastating disease. Compounds that stimulate the immune system to clear Aß may therefore have great therapeutic potential in AD patients. Monophosphoryl lipid A (MPL) is an LPS-derived Toll-like receptor 4 agonist that exhibits unique immunomodulatory properties at doses that are nonpyrogenic. We show here that repeated systemic injections of MPL, but not LPS, significantly improved AD-related pathology in APP(swe)/PS1 mice. MPL treatment led to a significant reduction in Aß load in the brain of these mice, as well as enhanced cognitive function. MPL induced a potent phagocytic response by microglia while triggering a moderate inflammatory reaction. Our data suggest that the Toll-like receptor 4 agonist MPL may be a treatment for AD.
Subject(s)
Alzheimer Disease/prevention & control , Brain/drug effects , Lipid A/analogs & derivatives , Toll-Like Receptor 4/agonists , Alzheimer Disease/metabolism , Alzheimer Disease/pathology , Amyloid beta-Protein Precursor/genetics , Amyloid beta-Protein Precursor/metabolism , Animals , Blotting, Western , Brain/metabolism , Brain/pathology , Cell Line , Cytokines/genetics , Cytokines/metabolism , Gene Expression/drug effects , HEK293 Cells , Humans , Immunity, Innate/drug effects , Ligands , Lipid A/administration & dosage , Lipid A/therapeutic use , Lipopolysaccharides/pharmacology , Mice , Mice, Inbred C57BL , Mice, Transgenic , Microglia/cytology , Microglia/drug effects , Microglia/metabolism , Microscopy, Fluorescence , Phagocytosis/drug effects , Presenilin-1/genetics , Presenilin-1/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Toll-Like Receptor 4/metabolismABSTRACT
Nicotine replacement therapies (NRT) have limited success in smoking cessation. The efficacy of nicotine may be compromised by its main metabolite, cotinine. An anti-cotinine vaccine to remove this antagonism could enhance the efficacy of NRT. We show that cotinine is a weak nicotinic agonist and decreases responses to nicotine, consistent with antagonism through receptor desensitisation. trans-4-Thiol cotinine was coupled to tetanus toxoid, and rats immunised repeatedly. Vaccination raised antibodies specific for cotinine that do not recognise other metabolites or nicotine. Increased serum cotinine concentrations following nicotine administration indicate sequestration of cotinine by antibodies, encouraging further evaluation of this vaccine in behavioural models of nicotine addiction and relapse.
Subject(s)
Cotinine/antagonists & inhibitors , Cotinine/immunology , Nicotine/administration & dosage , Smoking Cessation/methods , Vaccines, Conjugate/isolation & purification , Animals , Corpus Striatum/drug effects , Corpus Striatum/metabolism , Cotinine/metabolism , Dopamine/metabolism , Humans , In Vitro Techniques , Male , Nicotine/metabolism , Rats , Rats, Sprague-Dawley , Receptors, Nicotinic/drug effects , Receptors, Nicotinic/metabolismABSTRACT
BACKGROUND: Modified allergens with reduced IgE-binding activity represent an elegant approach to circumvent the risk of anaphylactic reactions in allergen-specific immunotherapy. OBJECTIVE: The current work investigated the effect of heat denaturation on the allergenic properties of recombinant ProDer p 1, a precursor form of the major house dust mite allergen Der p 1. METHODS: The IgE reactivity was estimated by direct and competition ELISA. The immunogenicity of heat-denatured ProDer p 1 was evaluated in naive and Der p 1-allergic mice. RESULTS: Heat denaturation in reducing conditions drastically reduced the in vitro ProDer p 1 IgE reactivity toward human allergic sera. In naive mice, heat-denatured ProDer p 1 generated mixed T(H)1-T(H)2 responses characterized by the absence of specific IgE with concomitant rise in specific IgG2a titers and the presence of IL-5 and IFN-gamma in splenocyte cultures. In contrast, natural Der p 1 or native ProDer p 1 induced typical strict T(H)2-biased allergic responses with strong IgG1 and IgE titers, whereas spleen cells exhibited only high IL-5 secretion. Moreover, native or heat-denatured ProDer p 1 vaccinations prevented airway eosinophil infiltrations after challenge. Although native or heat-treated ProDer p 1 adjuvanted with SBAS1b induced mixed T(H)1-T(H)2 responses in allergic mice, heat-denatured ProDer p 1, compared with native ProDer p 1, proved to be more effective in redirecting the T(H)2-allergic response toward T(H)1. CONCLUSION: Taken together, our results suggest that variants of Der p 1 with reduced IgE-binding reactivity could represent hypoallergenic molecules suitable for allergen-specific immunotherapy.
