Development of a sensitive phospho-p70 S6 kinase ELISA to quantify mTOR proliferation signal inhibition.

Abstract:

BACKGROUND:Drug blood levels can only serve as a surrogate because of the lack of information on the drug's direct pharmacological effects in the individual patient. Measurement of the mammalian target of rapamycin (mTOR) activity dependent on the phosphorylation status of p70 S6 kinase (p70 S6K) offers a practical way for monitoring pharmacodynamic drug activity, with the potential to better assess the state of immunosuppression in individual patients. MATERIAL AND METHODS:Here, we established a novel in vitro model system by treating Jurkat cells and peripheral blood mononuclear cells with different concentrations of sirolimus after stimulation with phorbol 12-myristate 13-acetate. RESULTS:A dose-dependent reduction of the p70 S6K phosphorylation status was demonstrated by Western blot and a newly established enzyme-linked immunosorbent assay (ELISA). Relative phospho-p70 S6K values from ELISA and relative densities from Western blot analysis in peripheral blood mononuclear cells revealed a strong correlation (Spearman correlation coefficient r s = 0.7, P = 0.01). Finally, parallel assays confirmed a sirolimus dose-dependent reduction of cytokine production and cell proliferation in the in vitro model. CONCLUSIONS:Pharmacodynamic monitoring of mTOR inhibition with a p70 S6K ELISA could guide mTOR inhibitor immunosuppression therapy toward a more individualized therapy. The usage of this technique now has to be evaluated in a clinical series of patients.

journal_name

Ther Drug Monit

authors

Hartmann B,He X,Keller F,Fischereder M,Guba M,Schmid H

doi

10.1097/FTD.0b013e3182804c9b

subject

Has Abstract

pub_date

2013-04-01 00:00:00

pages

233-9

issue

2

eissn

0163-4356

issn

1536-3694

pii

00007691-201304000-00013

journal_volume

35

pub_type

杂志文章
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