Format

Send to

Choose Destination
Alzheimers Dement. 2016 Oct;12(10):1098-1107. doi: 10.1016/j.jalz.2016.01.013. Epub 2016 Mar 23.

Internalization of tau antibody and pathological tau protein detected with a flow cytometry multiplexing approach.

Author information

1
Department of Neuroscience and Physiology, New York University School of Medicine, New York, NY, USA.
2
H. Lundbeck A/S, Valby, Denmark.
3
Department of Neuroscience and Physiology, New York University School of Medicine, New York, NY, USA; Department of Psychiatry, New York University School of Medicine, New York, NY 10016, USA. Electronic address: einar.sigurdsson@nyumc.org.

Abstract

INTRODUCTION:

Tau immunotherapy has emerged as a promising approach to clear tau aggregates from the brain. Our previous findings suggest that tau antibodies may act outside and within neurons to promote such clearance.

METHODS:

We have developed an approach using flow cytometry, a human neuroblastoma cell model overexpressing tau with the P301L mutation, and paired helical filament (PHF)-enriched pathologic tau to effectively screen uptake and retention of tau antibodies in conjunction with PHF.

RESULTS:

The flow cytometry approach correlates well with Western blot analysis to detect internalized antibodies in naïve and transfected SH-SY5Y cells (r2 = 0.958, and r2 = 0.968, P = .021 and P = .016, respectively). In transfected cells, more antibodies are taken up/retained as pathologic tau load increases, both under co-treated conditions and when the cells are pretreated with PHF before antibody administration (r2 = 0.999 and r2 = 0.999, P = .013 and P = .011, respectively).

DISCUSSION:

This approach allows rapid in vitro screening of antibody uptake and retention in conjunction with pathologic tau protein before more detailed studies in animals or other more complex model systems.

KEYWORDS:

Alzheimer's disease; Antibody; Flow cytometry; Immunotherapy; Internalization; Neuroblastoma cells; Paired helical filaments; SH-SY5Y cells; Tau protein; Uptake

PMID:
27016263
PMCID:
PMC5383206
DOI:
10.1016/j.jalz.2016.01.013
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center