Journal of Neurology

, Volume 253, Issue 9, pp 1177–1184 | Cite as

CSF protein profiling using Multiplex Immuno-assay

A potential new diagnostic tool for leptomeningeal metastases
  • Dieta Brandsma
  • Emile E. Voest
  • Wilco de Jager
  • Hans Bonfrer
  • Ale Algra
  • Willem Boogerd
  • Tiny Korse
  • Jaap C. Reijneveld
  • Marcel M. Verbeek
  • Ger Rijkers
  • Martin J.B. Taphoorn



The diagnosis of leptomeningeal metastases (LM) is based on clinical symptoms, magnetic resonance imaging (MRI) of brain and spine and cytological analysis of cerebrospinal fluid (CSF). The clinical picture of LM is highly variable and both cytological CSF analysis and contrast-enhanced MRI are limited in sensitivity. More sensitive tools are needed to diagnose LM. We measured a profile of proteins involved in adhesion and inflammation in the CSF of LM and control patients and determined their potential diagnostic value for LM.

Patients and methods

Using Multiplex Immuno-Assay (MIA), the CSF concentrations of nine soluble adhesion molecules, cyto- and chemokines were measured in patients with cytologically proven LM (n=57) and control patients with a systemic malignancy (n=20), aseptic/viral meningitis (n=11) or other (non-)neurological diseases (n=19).


We found high CSF levels of soluble Vascular Cell Adhesion Molecule-1 (sVCAM-1), soluble Intercellular Adhesion Molecule-1 (sICAM-1), Interleukin-8 (IL-8), Pulmonary and Activation Regulated Chemokine (PARC), Interleukin-18 (IL-18) and Interferon-γ inducible protein (IP-10) in patients with LM. The CSF protein profile in LM patients differed significantly from the profile found in control patients. Multivariate logistic regression and ROC analysis showed that the MIA-measured CSF protein profile has an additive discriminating value for LM above standard CSF parameters. A combination of total protein, glucose, IL-8, PARC and IP-10 CSF levels proved to be most discriminative between LM and non-LM patients.


Our results warrant a prospective study to determine whether a CSF protein profile, including IL-8, PARC and IP-10 has diagnostic value compared with CSF cytology, the golden standard for LM.


leptomeningeal metastases cerebrospinal fluid multiplex immuno-assay soluble adhesion molecules cytokines 


  1. 1.
    Aggarwal BB, Vilcek J (1991) Tumor Necrosis Factors: structure, function and mechanism of action. Dekker New YorkGoogle Scholar
  2. 2.
    Brandsma D, Reijneveld JC, Taphoorn MJ, de Boer HC, Gebbink MF, Ulfman LH, Zwaginga JJ, Voest EE (2002) Vascular cell adhesion molecule-1 is a key adhesion molecule in melanoma cell adhesion to the leptomeninges. Lab Invest 82:1493–1502PubMedGoogle Scholar
  3. 3.
    de Jager W, Prakken BJ, Bijlsma JW, Kuis W, Rijkers GT (2005) Improved multiplex immunoassay performance in human plasma and synovial fluid following removal of interfering heterophilic antibodies. J Immunol Methods 300:124–135PubMedCrossRefGoogle Scholar
  4. 4.
    de Jager W, te Velthuis H, Prakken BJ, Kuis W, Rijkers GT (2003) Simultaneous detection of 15 human cytokines in a single sample of stimulated peripheral blood mononuclear cells. Clin Diagn Lab Immunol 10:133–139PubMedCrossRefGoogle Scholar
  5. 5.
    Demopoulos A (2004) Leptomeningeal metastases. Curr Neurol Neurosci Rep 4:196–204PubMedGoogle Scholar
  6. 6.
    Fitzgerald SP, Lamont JV, McConnell RI, Benchikh el O (2005) Development of a high-throughput automated analyzer using biochip array technology. Clin Chem 51:1165–1176PubMedCrossRefGoogle Scholar
  7. 7.
    Freilich RJ, Krol G, DeAngelis LM (1995) Neuroimaging and cerebrospinal fluid cytology in the diagnosis of leptomeningeal metastasis. Ann Neurol 38:51–57PubMedCrossRefGoogle Scholar
  8. 8.
    Gerard C, Rollins BJ (2001) Chemokines and disease. Nat Immunol 2:108–115PubMedCrossRefGoogle Scholar
  9. 9.
    Gilburd B, Abu-Shakra M, Shoenfeld Y, Giordano A, Bocci EB, delle Monache F, Gerli R (2004) Autoantibodies profile in the sera of patients with Sjogren’s syndrome: the ANA evaluation–a homogeneous, multiplexed system. Clin Dev Immunol 11:53–56PubMedCrossRefGoogle Scholar
  10. 10.
    Hanley JA, McNeil BJ (1982) The meaning and use of the area under a receiver operating characteristic (ROC) curve. Radiology 143:29–36PubMedGoogle Scholar
  11. 11.
    Herrlinger U, Wiendl H, Renninger M, Forschler H, Dichgans J, Weller M (2004) Vascular endothelial growth factor (VEGF) in leptomeningeal metastasis: diagnostic and prognostic value. Br J Cancer 91:19–224Google Scholar
  12. 12.
    Hieshima K., Imai T, Baba M, Shoudai K, Ishizuka K, Nakagawa T, Tsuruta J, Takeya M, Sakaki Y, Takatsuki K, Miura R, Opdenakker G, Van Damme J, Yoshie O, Nomiyama H (1997) A novel human CC chemokine PARC that is most homologous to macrophage-inflammatory protein-1 alpha/LD78 alpha and chemotactic for T lymphocytes, but not for m-onocytes. J Immunol 159:1140–1149PubMedGoogle Scholar
  13. 13.
    Kaplan JG, DeSouza TG, Farkash A, Shafran B, Pack D, Rehman F, Fuks J, Portenoy R (1990) Leptomeningeal metastases: comparison of clinical features and laboratory data of solid tumors, lymphomas and leukemias. J Neurooncol 9:225–229PubMedCrossRefGoogle Scholar
  14. 14.
    Martin DB, Nelson PS (2001) From genomics to proteomics: techniques and applications in cancer research. Trends Cell Biol 11:S60–S65PubMedGoogle Scholar
  15. 15.
    Neville LF, Mathiak G, Bagasra O (1997) The immunobiology of interferon-gamma inducible protein 10 kD (IP-10): a novel, pleiotropic member of the C-X-C chemokine superfamily. Cytokine Growth Factor Rev 8:207–219PubMedCrossRefGoogle Scholar
  16. 16.
    Oppenheim JJ, Zachariae CO, Mukaida N, Matsushima K (1991) Properties of the novel proinflammatory supergene “intercrine” cytokine family. Annu Rev Immunol 9:617–648PubMedGoogle Scholar
  17. 17.
    Pestka S, Langer JA, Zoon KC, Samuel CE (1987) Interferons and their actions. Annu Rev Biochem 56:727–777PubMedCrossRefGoogle Scholar
  18. 18.
    Prabhakar U, Eirikis E, Davis HM (2002) Simultaneous quantification of proinflammatory cytokines in human plasma using the LabMAP assay. J Immunol Methods 260:207–218PubMedCrossRefGoogle Scholar
  19. 19.
    Reijneveld JC, Brandsma D, Boogerd W, Bonfrer JGM, Kalmijn S, Voest EE, Geurts-Moespot A, Visser MC, Taphoorn MJ (2005) CSF levels of angiogenesis related proteins in patients with leptomeningeal metastases. Neurology 65:1120–1122PubMedCrossRefGoogle Scholar
  20. 20.
    Scrideli CA, Queiroz RP, Takayanagui OM, Bernardes JE, Tone LG (2003) Polymerase chain reaction on cerebrospinal fluid cells in suspected leptomeningeal involvement in childhood acute lymphoblastic leukemia: comparison to cytomorphological analysis. Diagn Mol Pathol 12:124–127PubMedGoogle Scholar
  21. 21.
    Singh SK, Leeds NE, Ginsberg LE (2002) MR imaging of leptomeningeal metastases: comparison of three sequences. Am J Neuroradiol 23:817–821PubMedGoogle Scholar
  22. 22.
    Stockhammer G, Poewe W, Burgstaller S, Deisenhammer F, Muigg A, Kiechl S, Schmutzhard E, Maier H, Felber S, Schumacher P, Gunsilius E, Gastl G (2000) Vascular endothelial growth factor in CSF: a biological marker for carcinomatous meningitis. Neurology 54:1670–1676PubMedGoogle Scholar
  23. 23.
    Subira D, Castanon S, Roman A, Aceituno E, Jimenez-Garofano C, Jimenez A, Garcia R, Bernacer M (2001) Flow cytometry and the study of central nervous disease in patients with acute leukaemia. Br J Haematol 112:381–384PubMedCrossRefGoogle Scholar
  24. 24.
    Tallman RD, Kimbrough SM, O’Brien JF, Goellner JR, Yanagihara T (1985) Assay for beta-glucuronidase in cerebrospinal fluid: usefulness for the detection of neoplastic meningitis. Mayo Clin Proc 60:293–298PubMedGoogle Scholar
  25. 25.
    Twijnstra A, Nooyen WJ, van Zanten AP, Hart AA, Ongerboer de V (1986) Cerebrospinal fluid carcinoembryonic antigen in patients with metastatic and nonmetastatic neurological diseases. Arch Neurol 43:269–272PubMedGoogle Scholar
  26. 26.
    van Oostenbrugge RJ, Twijnstra A (1999) Presenting features and value of diagnostic procedures in leptomeningeal metastases. Neurology 53:382–385PubMedGoogle Scholar
  27. 27.
    Wasserstrom WR, Glass JP, Posner JB (1982) Diagnosis and treatment of leptomeningeal metastases from solid tumors: experience with 90 patients. Cancer 49:759–772PubMedCrossRefGoogle Scholar
  28. 28.
    Wasserstrom WR, Schwartz MK, Fleisher M, Posner JB (1981) Cerebrospinal fluid biochemical markers in central nervous system tumors: a review. Ann Clin Lab Sci 11:239–251PubMedGoogle Scholar
  29. 29.
    Weller M, Stevens A, Sommer N, Schabet M, Wietholter H (1992) Tumor cell dissemination triggers an intrathecal immune response in neoplastic meningitis. Cancer 69:1475–1480PubMedCrossRefGoogle Scholar
  30. 30.
    Xie K (2001) Interleukin-8 and human cancer biology. Cytokine Growth Factor Rev 12:375–391PubMedCrossRefGoogle Scholar

Copyright information

© Steinkopff Verlag Darmstadt 2006

Authors and Affiliations

  • Dieta Brandsma
    • 1
    • 2
  • Emile E. Voest
    • 2
  • Wilco de Jager
    • 3
  • Hans Bonfrer
    • 4
  • Ale Algra
    • 1
    • 5
  • Willem Boogerd
    • 6
  • Tiny Korse
    • 4
  • Jaap C. Reijneveld
    • 7
  • Marcel M. Verbeek
    • 8
  • Ger Rijkers
    • 3
  • Martin J.B. Taphoorn
    • 1
    • 9
  1. 1.Dept. of Neurology, G03.228University Medical Center UtrechtUtrechtThe Netherlands
  2. 2.Laboratory of Medical Oncology, Dept. of Medical OncologyUniversity Medical Center UtrechtUtrechtThe Netherlands
  3. 3.Dept. of Pediatric Immunology, Wilhelmina Children’s Hospital, University Medical Center UtrechtIACOPO Institute for Translational MedicineUtrechtThe Netherlands
  4. 4.Dept. of Clinical ChemistryAntoni van Leeuwenhoek HospitalAmsterdamThe Netherlands
  5. 5.Dept. of Clinical EpidemiologyJulius Center for Health Sciences and Primary Care, University Medical Center UtrechtUtrechtThe Netherlands
  6. 6.Dept. of NeurologyAntoni van Leeuwenhoek HospitalAmsterdamThe Netherlands
  7. 7.Dept. of NeurologyVU University Medical CenterAmsterdamThe Netherlands
  8. 8.Dept. of NeurologyUniversity Medical CenterNijmegenThe Netherlands
  9. 9.Dept. of NeurologyMedical Center HaaglandenDen HaagThe Netherlands

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