Journal of Neurology

, Volume 265, Issue 3, pp 510–521 | Cite as

CSF neurofilament proteins as diagnostic and prognostic biomarkers for amyotrophic lateral sclerosis

  • Daniela Rossi
  • Paolo Volanti
  • Liliana Brambilla
  • Tiziana Colletti
  • Rossella Spataro
  • Vincenzo La BellaEmail author
Original Communication


Elevated cerebrospinal fluid (CSF), Neurofilament Light (NF-L) and phosphorylated Heavy (pNF-H) chain levels have been found in Amyotrophic Lateral Sclerosis (ALS), with studies reporting a correlation of both neurofilaments (NFs) with the disease progression. Here, we measured NF-L and pNF-H concentrations in the CSF of ALS patients from a single tertiary Center and investigated their relationship with disease-related variables. A total of 190 ALS patients (Bulbar, 29.9%; Spinal, 70.1%; M/F = 1.53) and 130 controls with mixed neurological diseases were recruited. Demographic and clinical variables were recorded, and ΔFS was used to rate the disease progression. Controls were divided into two cohorts: (1) patients with non-inflammatory neurological diseases (CTL-1); (2) patients with acute/subacute inflammatory diseases and tumors, expected to lead to significant axonal and tissue damage (CTL-2). For each patient and control, CSF was taken at the time of the diagnostic work-up and stored following the published guidelines. CSF NF-L and pNF-H were assayed with commercially available ELISA-based methods. Standard curves (from independent ELISA kits) were highly reproducible for both NFs, with a coefficient of variation < 20%. We found that CSF NF-L and pNF-H levels in ALS were significantly increased when compared to CTL-1 (NF-L: ALS, 4.7 ng/ml vs CTL-1, 0.61 ng/ml, p < 0.001; pNF-H: ALS, 1.7 ng/ml vs CTL-1, 0.03 ng/ml, p < 0.0001), but not to CTL-2. Analysis of different clinical and prognostic variables disclosed meaningful correlations with both NF-L and pNF-H levels. Our results, from a relatively large ALS cohort, confirm that CSF NF-L and pNF-H represent valuable diagnostic and prognostic biomarkers in ALS.


ALS Neurofilaments NF-L pNF-H Disease progression CSF 



This work was supported by funds from the Italian Ministry of Health for the Ricerca Corrente to IRCCS Istituti Clinici Scientifici Maugeri SpA SB.

Compliance with ethical standards

Conflicts of interest

On behalf of all authors, the corresponding author states that there is no conflict of interest related to the Ms JOON-D-17-01506.

Supplementary material

415_2017_8730_MOESM1_ESM.docx (132 kb)
Supplementary material 1 (DOCX 132 kb)


  1. 1.
    Lee MK, Cleveland DW (1996) Neuronal intermediate filaments. Annu Rev Neurosci 19:187–217CrossRefPubMedGoogle Scholar
  2. 2.
    Petzold A (2005) neurofilament phosphoforms: surrogate markers of axonal injury, degeneration and loss. J Neurol Sci 233:183–198CrossRefPubMedGoogle Scholar
  3. 3.
    Rosengren LE, Karlsson J-E, Karlsson J-O et al (1996) Patients with amyotrophic lateral sclerosis and other neurodegenerative diseases have increased levels of neurofilament protein in CSF. J Neurochem 67:2013–2018CrossRefPubMedGoogle Scholar
  4. 4.
    Hu YY, He SS, Wang XC et al (2002) Elevated levels of phosphorylated neurofilament proteins in cerebrospinal fluid of Alzheimer disease patients. Neurosci Lett 320:156–160CrossRefPubMedGoogle Scholar
  5. 5.
    Pijnenburg YAL, Janssen JC, Schoonenboom NSM et al (2007) CSF neurofilaments in frontotemporal dementia compared to early onset Alzheimer’s disease and controls. Dement Geriatric Cogn Dis 23:225–230CrossRefGoogle Scholar
  6. 6.
    Mattsson N, Rüetschi U, Pijnenburg YAL et al (2008) Novel cerebrospinal fluid biomarkers of axonal degeneration in frontotemporal dementia. Mol Med Rep 1:757–761PubMedGoogle Scholar
  7. 7.
    Ganesalingam J, An J, Bowser R et al (2013) pNfH is a promising biomarker in ALS. Amyotrophic Lat Scler Frontotemp Degen 14:146–149CrossRefGoogle Scholar
  8. 8.
    Landqvist Waldö M, Santillo AF, Passant U et al (2013) Cerebrospinal fluid neurofilament light chain protein levels in subtypes of frontotemporal dementia. BMC Neurology 13:54e1–54e8CrossRefGoogle Scholar
  9. 9.
    Scherling CS, Hall T, Berisha F et al (2014) Cerebrospinal fluid neurofilament concentration reflects disease severity in frontotemporal degeneration. Ann Neurol 75:116–126CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Abdelhak A, Junker A, Brettschneider J et al (2015) Brain-specific cytoskeletal markers in cerebrospinal fluid: is there a common pattern between amyotrophic lateral sclerosis and primary progressive multiple sclerosis? Int J Mol Sci 16:17565–17588CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Xu Z, Henderson RD, David M, McCombe PA (2016) Neurofilaments as biomarkers for amyotrophic lateral sclerosis: a systematic review and meta-analysis. PLoS ONE 11(e0164625):e1–e18Google Scholar
  12. 12.
    Meeter LH, Dopper EG, Jiskoot LC et al (2016) Neurofilament light chain: a biomarker for genetic frontotemporal dementia. Ann Clin Transl Neurol 3:623–626CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Wilke C, Preische O, Deuschle C et al (2016) Neurofilament light chain in FTD is elevated not only in cerebrospinal fluid, but also in serum. J Neurol Neurosurg Psychiatry 87:1270–1272CrossRefPubMedGoogle Scholar
  14. 14.
    Mattsson N, Andreasson U, Zettenberg H et al (2017) association of plasma neurofilament light with neurodegeneration in patients with Alzheimer disease. JAMA Neurol 74:557–566CrossRefPubMedGoogle Scholar
  15. 15.
    Hu X, Yang Y, Gong D (2017) Cerebrospinal fluid levels of neurofilament light chain in multiple system atrophy relative to Parkinson’s disease: a meta-analysis. Neurol Sci 38:407–414CrossRefPubMedGoogle Scholar
  16. 16.
    Hansson O, Janelidze S, Hall S et al (2017) Blood-based NfL. A biomarker for differential diagnosis of parkinsonian disorder. Neurology 88:930–937CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Krut JJ, Mellberg T, Price RW et al (2014) Biomarker evidence of axonal injury in neuroasymptomatic HIV-1 patients. PLoS ONE 9(e88951):e1–e7Google Scholar
  18. 18.
    Spataro R, Lo Re M, Piccoli T et al (2010) Causes and place of death in Italian patients with Amyotrophic Lateral Sclerosis. Acta Neurol Scand 122:217–223CrossRefPubMedGoogle Scholar
  19. 19.
    Traynor BJ, Codd MB, Corr B et al (2000) Amyotrophic lateral sclerosis mimic syndromes: a population-based study. Arch Neurol 57:109–113CrossRefPubMedGoogle Scholar
  20. 20.
    Brooks BR, Miller RG, Swash M, Munsat TL, World Federation of Neurology Research Group on Motor Neuron Diseases (2000) El Escorial revisited: revised criteria for the diagnosis of amyotrophic lateral sclerosis. Amyotroph Lateral Scler 1:293–299Google Scholar
  21. 21.
    Cedarbaum JM, Stambler N, Malta E et al (1999) The ALSFRS-R: a revised ALS functional rating scale that incorporates assessments of respiratory function. BDNF ALS Study Group (Phase III). J Neurol Sci 169:13–21CrossRefPubMedGoogle Scholar
  22. 22.
    Menke RAL, Gray E, Lu C-H et al (2015) CSF neurofilament light chain reflects corticospinal tract degeneration in ALS. Ann Clin Transl Neurol 2:748–755CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Reijn TS, Abdo WF, Schelhaas HJ, Verbeek MM (2009) CSF neurofilament protein analysis in the differential diagnosis of ALS. J Neurol 256:615–619CrossRefPubMedGoogle Scholar
  24. 24.
    Tortelli R, Ruggieri M, Cortese R et al (2012) Elevated cerebrospinal fluid neurofilament light levels in patients with amyotrophic lateral sclerosis: a possible marker of disease severity and progression. Eur J Neurol 19:1561–1567CrossRefPubMedGoogle Scholar
  25. 25.
    Boylan KB, Glass JD, Crook JE et al (2013) Phosphorylated neurofilament heavy subunit (pNF-H) in peripheral blood and CSF as potential prognostic biomarker in amyotrophic lateral sclerosis. J Neurol Neurosurg Psychiatry 84:467–472CrossRefPubMedGoogle Scholar
  26. 26.
    Gaiottino J, Norgren N, Dobson R et al (2013) Increased neurofilament light chain blood levels in neurodegenerative neurological disease. PLoS ONE 8(e75091):e1–e9Google Scholar
  27. 27.
    Gonçalves M, Tillack L, de Carvalho M et al (2014) Phosphoneurofilament heravy chain and N-glycomics from the cerebrospinal fluid in amyotrophic lateral sclerosis. Clin Chim Acta 438:342–349CrossRefPubMedGoogle Scholar
  28. 28.
    Lu C-H, Petzold A, Topping J et al (2015) plasma neurofilament heavy chain levels and disease progression in amyotrophic lateral sclerosis: insights from a longitudinal study. J Neurol Neurosurg Psychiatry 86:565–573CrossRefPubMedGoogle Scholar
  29. 29.
    Lu C-H, Macdonald-Wallis C, Gray E et al (2015) Neurofilament light chain. A prognostic biomarker in amyotrophic lateral sclerosis. Neurology 84:2247–2257CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Weydt P, Oeckl P, Huss A et al (2016) Neurofilament levels as biomarkers in asymptomatic and symptomatic familial amyotrophic lateral sclerosis. Ann Neurol 79:152–158CrossRefPubMedGoogle Scholar
  31. 31.
    Steinacker P, Feneberg E, Weishaupt J et al (2016) Neurofilaments in the diagnosis of motoneuron diseases: a prospective study of 455 patients. J Neurol Neurosurg Psychiatry 87:12–20PubMedGoogle Scholar
  32. 32.
    Oeckl P, Jardel C, Salachas F et al (2016) Multicenter validation of CSF neurofilaments as diagnostic biomarkers for ALS. Amyotr Lat Scler Frontotemp Degen 17:404–413CrossRefGoogle Scholar
  33. 33.
    Steinacker P, Huss A, Mayer B et al (2017) diagnostic and prognostic significance of neurofilament light chain NF-L, but not progranulin and S100B, in the course of amyotrophic lateral sclerosis: data from the German MND-net. Amyotr Lat Scler Frontotemp Degen 18:112–119CrossRefGoogle Scholar
  34. 34.
    Gaiani A, Martinelli I, Bello L et al (2017) Diagnostic and prognostic biomarkers in amyotrophic lateral sclerosis. Neurofilament light chain levels in definite subtypes of disease. JAMA Neurol 74:525–532CrossRefPubMedGoogle Scholar
  35. 35.
    Poesen K, De Schaepdryver M, Stubendorff B et al (2017) neurofilaments markers for ALS correlate with extent of upper and lower motor neuron disease. Neurology 88:2302–2309CrossRefPubMedGoogle Scholar
  36. 36.
    Turner MR, Gray E (2015) Are neurofilaments heading for the ALS clinic? J Neurol Neurosurg Psychiatry 87:3–4PubMedGoogle Scholar
  37. 37.
    De Carvaho M, Dengler R, Eisen A et al (2008) Electrodiagnosis criteria for diagnosis of ALS. Clin Neurophysiol 119:497–503CrossRefGoogle Scholar
  38. 38.
    Neary D, Snowden JS, Gustafson L et al (1998) Frontotemporal lobar degeneration: a consensus on clinical diagnostic criteria. Neurology 51:1546–1554CrossRefPubMedGoogle Scholar
  39. 39.
    Bang J, Spina S, Miller BL (2015) Frontotemporal dementia. Lancet 386:1672–1682CrossRefPubMedGoogle Scholar
  40. 40.
    Kimura F, Fujimura C, Ishida S et al (2006) Progression rate of ALSFRS-R at the time of diagnosis predicts survival time in ALS. Neurology 66:265–267CrossRefPubMedGoogle Scholar
  41. 41.
    Cellura E, Spataro R, Taiello AC, La Bella V (2012) Factors affecting the diagnostic delay in amyotrophic lateral sclerosis. Clin Neurol Neurosurg 114:550–554CrossRefPubMedGoogle Scholar
  42. 42.
    Robinson KM, Lacey SC, Grugan P, Glosser G, Grossman M, McCluskey LF (2006) Cognitive functioning in sporadic amyotrophic lateral sclerosis: a six month longitudinal study. J Neurol Neurosurg Psychiatry 77:668–670CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Phukan J, Pender NP, Hardiman O (2007) Cognitive impairment in amyotrophic lateral sclerosis. Lancet Neurol 6:994–1003CrossRefPubMedGoogle Scholar
  44. 44.
    Kasper E, Zydatiss K, Schuster C, Machts J, Bittner D, Kaufmann J et al (2016) No change in executive performance in ALS patients: a longitudinal neuropsychological study. Neurodegener Dis 16:184–191CrossRefPubMedGoogle Scholar
  45. 45.
    Burkhardt C, Neuwirth C, Weber M (2017) Longitudinal assessment of the Edinburgh Cognitive and Behavioural Amyotrophic Lateral Sclerosis Screen (ECAS): lack of practice effect in ALS patients. Amyotroph Lateral Sclerosis Frontotemp Degener 18:202–209CrossRefGoogle Scholar
  46. 46.
    Teunissen CE, Petzold A, Bennett JL et al (2009) A consensus protocol for the standardization of cerebrospinal fluid collection and biobanking. Neurology 73:1914–1922CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Abdoa FW, Bloema BR, Van Geel WJ et al (2007) CSF neurofilament light chain and tau differentiate multiple system atrophy from Parkinson’s disease. Neurobiol Aging 28:742–747CrossRefGoogle Scholar
  48. 48.
    Steinacker P, Blennow K, Halbgebauer S et al (2016) Neurofilaments in blood and CSF for diagnosis and prediction of onset in Creutzfeldt-Jakob disease. Sci Rep 6:38737 e1–38737 e6CrossRefGoogle Scholar
  49. 49.
    Kang DX, Fan DS (1995) The electrophysiological study of differential diagnosis between amyotrophic lateral sclerosis and cervical spondylotic myelopathy. Electromyogr Clin Neurophysiol 35:231–238PubMedGoogle Scholar
  50. 50.
    Andersen PM, Abrahams S, Borasio GD et al (2012) EFNS guidelines on the Clinical Management of Amyotrophic Lateral Sclerosis (MALS) – revised report of an EFNS task force. Eur J Neurol 19:360–375CrossRefPubMedGoogle Scholar
  51. 51.
    Swinnen B, Robberecht W (2014) The phenotypic variability of amyotrophic lateral sclerosis. Nature Rev Neurol 10:661–670CrossRefGoogle Scholar
  52. 52.
    Appel V, Stewart SS, Smith RG, Appel SH (1987) A rating scale for amyotrophic lateral sclerosis: description and preliminary experience. Ann Neurol 22:328–333CrossRefPubMedGoogle Scholar
  53. 53.
    Mitsumoto H, Brooks BR, Silani V (2014) Clinical trials in amyotrophic lateral sclerosis: why so many negative trials and how can trials be improved? Lancet Neurol 13:1127–1138CrossRefPubMedGoogle Scholar
  54. 54.
    Bensimon G, Lacomblez L, Meininger V, for the ALS/Riluzole study group (1994) A controlled trial of riluzole in amyotrophic lateral sclerosis. New Engl J Med 330:585–591CrossRefPubMedGoogle Scholar
  55. 55.
    Elia AE, Lalli S, Monsurrò MR et al (2015) Tauroursodeoxycholic acid in the treatment of patients with amyotrophic lateral sclerosis. Eur J Neurol 23:45–52CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Laboratory for Research on Neurodegenerative DisordersICS MaugeriPaviaItaly
  2. 2.ALS Center and Neurorehabilitation UnitICS MaugeriMistrettaItaly
  3. 3.Department of Experimental BioMedicine and Clinical Neurosciences, ALS Clinical Research CenterUniversity of PalermoPalermoItaly

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