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Internalization of tau antibody and pathological tau protein detected with a flow cytometry multiplexing approach
Author(s) -
Shamir Dov B.,
Rosenqvist Nina,
Rasool Suhail,
Pedersen Jan T.,
Sigurdsson Einar M.
Publication year - 2016
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2016.01.013
Subject(s) - flow cytometry , internalization , antibody , western blot , transfection , microbiology and biotechnology , tau protein , chemistry , cytometry , immunotherapy , pathology , biology , cancer research , cell , immunology , cell culture , medicine , immune system , biochemistry , gene , genetics , disease , alzheimer's disease
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 (r 2 = 0.958, and r 2 = 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 (r 2 = 0.999 and r 2 = 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.