Plasma tau levels in Alzheimer's disease

  • Henrik Zetterberg1, 2Email author,

    Affiliated with

    • David Wilson3,

      Affiliated with

      • Ulf Andreasson1,

        Affiliated with

        • Lennart Minthon4,

          Affiliated with

          • Kaj Blennow1,

            Affiliated with

            • Jeffrey Randall3 and

              Affiliated with

              • Oskar Hansson4

                Affiliated with

                Alzheimer's Research & Therapy20135:9

                DOI: 10.1186/alzrt163

                Published: 28 March 2013

                Introduction

                Efforts to find reliable blood biomarkers for Alzheimer's disease (AD) in a highly warranted clinical laboratory test have met with little success. There is no clear change in plasma β-amyloid in AD, and assays for the axonal injury marker tau have been hampered by a lack of analytical sensitivity for accurate measurement in blood samples [1]. Here, the results of a novel ultra-sensitive assay for tau in peripheral blood are reported.

                Findings

                We have developed an ultra-sensitive assay for tau in peripheral blood [2]. In brief, the assay is based on digital array technology [3] and uses the Tau5 monoclonal antibody for capture (Covance, Princeton, NJ, USA) and HT7 and BT2 monoclonal antibodies for detection (Pierce, now part of Thermo Fisher Scientific Inc., Waltham, MA, USA). This combination reacts with both normal and phosphorylated tau with epitopes in the mid-region of the molecule, making the assay sensitive to all known tau isoforms. The calibrator was recombinant tau 381 (EMD Millipore Corporation, Billerica, MA, USA). To minimize matrix effects, all samples were diluted 1:4 in phosphate-buffered saline with 2% bovine serum albumin diluent prior to assay. The limit of detection of the assay, which requires 30 μL of plasma, is 0.02 pg/mL [2], which is more than 1,000-fold more sensitive than conventional immunoassays.

                Here, we assess the association of plasma tau levels with AD in a cross-sectional study of 54 patients with AD dementia [4], 75 patients with mild cognitive impairment (MCI) [5], and 25 cognitively normal controls (Table 1). All participants were recruited at the specialized memory clinic at Skåne University Hospital in Malmö, Sweden, and underwent extensive clinical evaluation, including cerebrospinal fluid (CSF) sampling by lumbar puncture, in addition to venipuncture and collection of blood in ethylenediaminetetraacetic acid (EDTA) tubes for plasma preparation by centrifugation within 15 minutes from sampling. Plasma samples were aliquoted into cryo tubes and stored at -80°C pending analysis, which was performed on one occasion by using one batch of reagents with an average coefficient of variation of 9.7% for triplicate measurements of each sample. The patients with MCI were cognitively stable for an average of 101 months (n = 36) or developed AD dementia (n = 35) or other types of dementias - vascular dementia (n = 3) and semantic dementia (n = 1) - during follow-up. The study was approved by the regional ethics committee at Lund University and complied with the Declaration of Helsinki. Informed consent was obtained from all study participants.
                Table 1

                Demographic and biochemical data

                 

                AD (n = 54)

                MCI (n = 75)

                Controls (n = 25)

                Age, years

                75 (6.2)

                68 (9.3)

                74 (6.7)

                Gender, male/female

                17/37

                29/46

                6/19

                MMSE, score

                19 (4.9)a

                27 (1.6)

                29 (1.4)

                Plasma T-tau, pg/mL

                8.80 (10.1)b,c

                4.68 (4.25)

                4.43 (2.83)

                CSF T-tau, pg/mL

                828 (375)a

                550 (421)

                507 (254)

                CSF P-tau, pg/mL

                123 (49.2)a

                78.1 (28.8)

                73.4 (20.5)

                CSF Aβ42, pg/mL

                380 (87.6)a

                478 (204)

                584 (215)

                Quantitative data are presented as mean (standard deviation). Statistical differences were determined by using nonparametric tests. Cerebrospinal fluid (CSF) biomarker concentrations are INNOTEST ELISA-normalized Luminex AlzBio3 (Innogenetics, Gent, Belgium) values. aCompared with patients with mild cognitive impairment (MCI) or controls, P <0.001. bCompared with patients with MCI, P = 0.001. cCompared with controls, P = 0.02. AD, Alzheimer's disease; MMSE, mini-mental state examination.

                Tau levels in plasma were significantly higher in AD patients compared with both controls and MCI patients (Figure 1a). MCI patients who developed AD during follow-up had tau levels similar to those of patients with stable MCI and cognitively normal controls (Figure 1b). There was no correlation between tau levels in plasma and CSF in any diagnostic group (Figure 1c).
                http://static-content.springer.com/image/art%3A10.1186%2Falzrt163/MediaObjects/13195_2013_139_Fig1_HTML.jpg
                Figure 1

                Elevated tau levels in plasma from patients with Alzheimer's disease (AD). (a) Plasma levels of tau are elevated in patients with AD compared with cognitively normal controls and patients with mild cognitive impairment (MCI). (b) MCI patients who developed AD (MCI-AD) during follow-up had baseline tau levels similar to those of patients with stable MCI (SMCI). (c) There was no correlation between tau levels in plasma and cerebrospinal fluid (CSF) in any diagnostic group. Thin horizontal lines in panels (a) and (b) indicate medians. A nonparametric Kruskal-Wallis test followed by Mann-Whitney was performed to test for statistical significance. Spearman's rank correlation coefficient was used to assess the relationship between plasma and CSF tau concentrations in panel (c), where open circles, gray squares, and black triangles represent AD, MCI, and controls, respectively.

                The results of this study have several important implications. First, plasma tau levels are elevated in AD but with overlapping ranges across diagnostic groups. This overlap diminishes the utility of plasma tau as a diagnostic test. However, further studies are needed to evaluate plasma tau as a first-in-line screening tool (for example, in the primary care setting and perhaps together with other markers in a biomarker panel). Second, normal plasma tau levels in the MCI stage of AD suggest that plasma tau is a late marker, requiring substantial axonal injury before increasing to abnormal levels. In this context, other neurodegenerative diseases (for example, Creutzfeldt-Jakob disease) as well as acute conditions (for example, stroke and brain trauma) should be tested. Third, the lack of correlation of tau levels in plasma and CSF suggests that steady-state concentrations of tau in these two body fluids are differentially regulated. In our earlier study of patients with hypoxic brain injury following cardiac arrest, tau was rapidly (within 24 hours) cleared from blood in patients with good neurological outcome [2], indicating potent clearance mechanisms for this marker in the bloodstream. This may obscure any correlation with CSF tau levels, which stay elevated for weeks following an acute neurological insult [6].

                Abbreviations

                AD: 

                Alzheimer's disease

                CSF: 

                cerebrospinal fluid

                MCI: 

                mild cognitive impairment.

                Declarations

                Acknowledgements

                This study was funded by grants from Swedish Brain Power, the Swedish Research Council, the Wolfson Foundation, the Alzheimer's Association, the JPND Project BIOMARKAPD, Swedish State Support for Clinical Research, the Swedish Brain Fund, the Alzheimer Foundation of Sweden, and the Dementia Association of Sweden.

                Authors’ Affiliations

                (1)
                Clinical Neurochemistry Laboratory, Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry, the Sahlgrenska Academy at the University of Gothenburg
                (2)
                UCL Institute of Neurology
                (3)
                Quanterix Corporation
                (4)
                Clinical Memory Research Unit, Clinical Sciences Malmö, Lund University

                References

                1. Blennow K, Hampel H, Weiner M, Zetterberg H: Cerebrospinal fluid and plasma biomarkers in Alzheimer disease. Nat Rev Neurol 2010, 6:131–144.PubMedView Article
                2. Randall J, Mörtberg E, Provuncher GK, Fournier DR, Duffy DC, Rubertsson S, Blennow K, Zetterberg H, Wilson DH: Tau proteins in serum predict neurological outcome after hypoxic brain injury from cardiac arrest: results of a pilot study. Resuscitation 2012, in press. Aug 9
                3. Rissin DM, Kan CW, Campbell TG, Howes SC, Fournier DR, Song L, Piech T, Patel PP, Chang L, Rivnak AJ, Ferrell EP, Randall JD, Provuncher GK, Walt DR, Duffy DC: Single-molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations. Nat Biotechnol 2010, 28:595–599.PubMedView Article
                4. McKhann G, Drachman D, Folstein M, Katzman R, Price D, Stadlan EM: Clinical diagnosis of Alzheimer's disease: report of the NINCDS-ADRDA Work Group under the auspices of Department of Health and Human Services Task Force on Alzheimer's Disease. Neurology 1984, 34:939–944.PubMedView Article
                5. Winblad B, Palmer K, Kivipelto M, Jelic V, Fratiglioni L, Wahlund LO, Nordberg A, Bäckman L, Albert M, Almkvist O, Arai H, Basun H, Blennow K, de Leon M, DeCarli C, Erkinjuntti T, Giacobini E, Graff C, Hardy J, Jack C, Jorm A, Ritchie K, van Duijn C, Visser P, Petersen RC: Mild cognitive impairment - beyond controversies, towards a consensus: report of the International Working Group on Mild Cognitive Impairment. J Intern Med 2004, 256:240–246.PubMedView Article
                6. Hesse C, Rosengren L, Andreasen N, Davidsson P, Vanderstichele H, Vanmechelen E, Blennow K: Transient increase in total tau but not phospho-tau in human cerebrospinal fluid after acute stroke. Neurosci Lett 2001, 297:187–190.PubMedView Article

                Copyright

                © BioMed Central Ltd 2013

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