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Detecting immunoglobulin abnormalities in the cerebrospinal fluid (CSF): Which methods are best for clinical purposes?

  • Laurenzi M. A. 
  • Provinciali L. 
  • Giovagnoli A. R. 
  • Busco
  • Bocchini V. 
  • Paris L. 
  • Chiodi F. 
Original Articles

Abstract

Several techniques are now available for the quantitative and qualitative examination of CSF proteins modified during the course of neurological disease. The CSF and serum of 42 patients suffering from various neurological diseases and of 7 controls were examined using quantitative methods-single radial immunodiffusion, nephelometric analysis-and qualitative methods—(i) isoelectric-focusing on polyacrylamide gel and (ii) on agarose gel, isoelectric focusing of unconcentrated CSF followed by transfer to cellulose nitrate membranes and (iii) immunoperoxidase staining and (iv) double antibody peroxidase labeling and avidin-biotin amplification. In the present study the results and the advantages of different methods are described and compared.

Key-Words

CSF immunoglobulins quantitative and qualitative methods single radial immunodiffusion nephelometric analysis-isoelectric focusing on polyaerylamide and agarose gelimmunoperoxidase staining avidin-biotin amplification 

Sommario

Numerose metodiche sono attualmente disponibili per l'esame quantitativo e qualitativo delle proteine liquorali che risultano alterate in numerose malattie neurologiche.

Liquor e siero di 42 pazienti affetti da diverse malattie neurologiche e di 7 controlli sono stati esaminati con metodiche quantitative — immunodiffusione radiale, analisi nefelometrica — e qualitative — (i) isoelettro focusing su poliacrilamide e (ii) su agarosio, isoelettrofocusing di liquor non concentrato seguito da trasferimento su membrane di nitrato di cellulosa e (iii) colorazione con immunoperossidasi e (iv) doppia definizione con perossidasi ed anticorpi seguito da amplificazione con avidin-biotina.

I risultati ed i vantaggi di ogni metodica sono descritti e confrontati tra loro.

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References

  1. [1]
    Laterre E.C., Callewaeri A., Heremans J.F., Sfaello Z.:Electrophoretic morphology of gamma globulins in cerebrospinal fluid of multiple sclerosis and other diseas of the nervous system. Neurology, 20:982–990. 1970.PubMedGoogle Scholar
  2. [2]
    Link H., Muller R.:Immnoglobulins in multiple sclerosis and infections of the nervous system. Arch Neurol. 25:326–344, 1971.PubMedGoogle Scholar
  3. [3]
    Vandvik B., Skrede S.:Electrophoretic examination of cerebrospinal fluid proteins in multiple sclerosis and other neurological diseases. Europ. Neurol., 9:224–241, 1973.PubMedGoogle Scholar
  4. [4]
    Karcher D., Matthyssens G., Lowenthal A.:Isolation and characterization of IgG globulins in subacute sclerosing panencephalitis. Immunology, 23:93–99, 1972.PubMedGoogle Scholar
  5. [5]
    Link H., Panelius M., Salmi A.A.:Immonuglobulins and measles antibodies in subacute sclerosing panecephalitis. Arch. Neurol. 28:23–30, 1973.PubMedGoogle Scholar
  6. [6]
    Vandvik B., Weil M.L., Grandien M., Norrby E.:Progressive rubella virus panencephalitis: Synthesis of oligoclonal virus-specific IgG antibodies and homogeneous free light chains in the central nervous system. Acta neurol. scand., 57:63–64, 1978.Google Scholar
  7. [7]
    Link H., Norrby E., Olsson J.E.:Immonuglobulins and measles antibodies in optic neuritis. N. Engl. J. Med., 289:1103–1107, 1973.PubMedGoogle Scholar
  8. [8]
    Nikoskelainen E., Irjala K., Salmi A.:Cerebrospinal fluid findings in patients with optic neuritis. Acta Ophtalmol., 53:105–119, 1975.Google Scholar
  9. [9]
    Link H., Norrby E., Olsson J.E.:Immonuglobulins abnormalities and measles antibody response in chronic myelopathy. Arch. Neurol. 33:26–32, 1976.PubMedGoogle Scholar
  10. [10]
    Paty D.W., Blume W.T., Beown W.F., Jaatoul N., Kertesz A., Mcinnis W.:Chronic progressive myelopathy: Investigation with CSF electrophoresis, evoked potentials, and CAT scan. Ann. Neurol. 6:419–424, 1979.CrossRefPubMedGoogle Scholar
  11. [11]
    Adornato B.T., Houff S.A., Engel W.K., Dalakas M., Madden D.L., Sever J.L.:Abnormal immunogloblin bands in cerebrospinal fluid in myasthenia gravis. Lancet, 2:367–368, 1978.PubMedGoogle Scholar
  12. [12]
    Fryden A., Link H., Norrby E.:Cerebrospinal fluid and serum immunoglobulins and antibody titers in mumps meningits and aseptic mengitis of other etiology. Infect. Immn. 21:852–961, 1978.Google Scholar
  13. [13]
    Skoldenberg B., Carlstrom A. Forsgren M., Norrby E.:Transient appearence of oligoclonal immunoglobulins and measles virus antibodies in cerebrospinal fluid in a case of acute measles encephalitis. Clin. Exp. Immunol. 23:451–455, 1976.PubMedGoogle Scholar
  14. [14]
    Link H.:Demonstration of oligoclonal immunoglobulin G in Guillain-Barré syndrome. Acta neurol. scand., 52:111–120, 1975.PubMedGoogle Scholar
  15. [15]
    Link H., Wahren B., Norrby E.:Pleocytosis and imunoglobulin changes in cerebrospinal fluid and herpesvirus serology in patients with Guillan-Barré syndrome. J. clin. Microbiol., 9:305–316, 1979.PubMedGoogle Scholar
  16. [16]
    Rostrom B., Link H.:Oligoclonal immunoglobulins in cerebrospinal fluid in acute cerebrovascular diseses. Neurology, 31:590–596, 1981.PubMedGoogle Scholar
  17. [17]
    Vandvik B., Norrby E.:Oligoclonal IgG antibody response in the central nervous system to different measles virus antigens in subacute sclerosing panencephalitis. Proc. nat. Acad. Sci., 70:1060–1063, 1973.PubMedGoogle Scholar
  18. [18]
    Laurenzi M.A., Link H.:Comparison between agarose gel electrophoresis and isoelectric focusing of CSF for demonstration of oligoclonal imunoglobulin bands in neurological disorders. Acta Neurol. Scand, 58:148–156, 1978.PubMedGoogle Scholar
  19. [19]
    Kjellin K.G., Stibler H.:Protein patterns of CSF in hereditary ataxias and hereditary spastic paraplegia. J. neurol. sci., 25:65–74, 1975.PubMedGoogle Scholar
  20. [20]
    Kjellin K.G., Stibler H.:Isoelectric focusing and electrophoresis of CSF proteins in muscolar dystrophies and spinal muscolar atrophies. J. Neurol. Sci., 27:45–57, 1976.CrossRefPubMedGoogle Scholar
  21. [21]
    Kjellin K.G., Stibler H.:CSF-protein pattern in extrapyramidal diseases. Europ. Neurol., 12: 186–194, 1974.PubMedGoogle Scholar
  22. [22]
    Mancini G., Carbonara A.O., Heremans J.F.:Immunochemical quantitation of antigens by single radial imunodifussion. Immunochemistry, 2: 235–254, 1965.CrossRefPubMedGoogle Scholar
  23. [23]
    Tibbling G., Link H., Ohman S.:Principles of albumin and IgG analyses in neurological disorders. I. Establishment of reference values. Scand. J. clin. Lab. Invest. 37:385–390, 1977.PubMedGoogle Scholar
  24. [24]
    Delmotte P.:Gel isoelectric focusing of cerebrospinal fluid proteins: a potential diagnostic tool. Z Klin. Chem. Klin. Biochem. 9:334–336, 1971.PubMedGoogle Scholar
  25. [25]
    Vesterberg O.:Isoelectric focusing of proteins in polyacrylamide gels. Biochim. biophys. Acta 257:11–19, 1972.PubMedGoogle Scholar
  26. [26]
    Delmotte P.:Etude par focalisation isoélectrique des gamma globulines du liquide céphalo-rachiden dans la sclérose en plaques et d'autres maladies neurologiques. Université de Poities, UER Sciences, thèse n. 38. 1975.Google Scholar
  27. [27]
    Kjellin K.J., Siden A.:Aberrant CSF protein fractions found by electrofocusing in multiple sclerosis. Europ. Neurol., 15:40–50, 1977.PubMedGoogle Scholar
  28. [28]
    Trotter J.L., Banks G. Wang P.:Isoelectric focusing of gamma globulins in cerebrospinal fluid from patients with multiple sclerosis. Clin. Chem., 23:2213–2215, 1977.PubMedGoogle Scholar
  29. [29]
    Olsson J.E., Nelsson K.:Gamma globulins of CSF and serum in multiple sclerosis: isoelectric focusing on polyacrylamide gel and agar gel electrophoresis. Neurology 29:1383–1391, 1979.Google Scholar
  30. [30]
    Laurenzi M.A.:Immunochemical characterization of immunoglobulins and viral antibodies synthesized within the central nervous system in patients with multiple sclerosis and controls. Acta Neurol. Scand., suppl. 84, vol. 63, 1–84, 1981.Google Scholar
  31. [31]
    Savaris C.A., Zamcheck N.:Isoelectric focusing in agarose. J. Immunol. Methods. 63, 2991–95, 1975.Google Scholar
  32. [32]
    Link H., Kostulas V.:Utility of isoelectric focusing of cerebrospinal fluid and serum on agarose evaluated for neurological patients. Clin. Chem. 29: 810–815, 1983.PubMedGoogle Scholar
  33. [33]
    Towbin H. Staehelin T., Gordon J.:Electrophoretic transfer of proteins from polyacrilamide gels to mitrocellulosa sheets; procedure and some applications. Proc. Natl. Acad. Sci., USA, 76:4350–4354, 1979.PubMedGoogle Scholar
  34. [34]
    Walker R.W.M., Keir G. Johnson M.H., Thompson E.J.:A rapid method for detecting oligoclonal IgG in unconcentrated CSF by agarose isoelectric focusing, transfer to cellulose nitrate and immunoperoxidase staining. J. Neuroimmunol., 4:141–148, 1983.CrossRefPubMedGoogle Scholar
  35. [35]
    Olsson T., Kostulas V. Link H.:Improved detection of oligoclonal IgG in cerebrospinal fluid by isolectric focusing in agarose double-antibody peroxidase labeling and Avidin-Biotin amplification. Clin. Chem., 30:1243–1249, 1984.Google Scholar
  36. [36]
    Forsberg P., Henriksson A.M., Link H., Ohman S.:Reference values for CSF-IgM, CSF-IgM:S-IgM ratio and IgM index and its application to patients with multiple sclerosis and aseptic meningoencephalitis. Scand. J. clin. Lab. Invest., 44:7–12, 1984.PubMedGoogle Scholar
  37. [37]
    Williams A.C., Mingioli E.S., Mcfarland H.F., Tourtellotte W.W., Mcfarlin D.E.:Increased CSF IgM in multiple sclerosis. Neurology, 28:996–998, 1978.Google Scholar
  38. [38]
    Sindic C., Cambioso C.L., Masson P.L., Laterre E.C.:Clinical relevance of the determination of IgM in the cerebrospinal fluid with special reference to multiple sclerosis patients. In Progress in Multiple Sclerosis. eds. H.J. Bauer, S. Poser, G. Ritter, pp. 176–179, Springer-Verlag, Berlin, 1980.Google Scholar
  39. [39]
    Rosen A.K., Ek P., Aman P.:Agarose isoelectric focusing of native human immunoglobulin M. ad alfa2 macroglobulin. J. Immunol. Meth. 63, 1–11, 1979.Google Scholar
  40. [40]
    Elkon K.B.:Isoelectric focusing of human IgA and secretory proteinss using thin layer agarose gels and mitrocellulose capillary blotting. J. Immunol. Meth., 25:313–321, 1984.Google Scholar
  41. [41]
    Ritchie R.F., Smith R.:Immunofixation. I. General principles and application to agarose gel. electrophoresis. Clin. Chem., 22:497–499, 1976.PubMedGoogle Scholar
  42. [42]
    Laurenzi M.A., Link H.:Characterization of the mobility on isoelectric focusing of individual proteins in CSF and serum by immunofixation. J. Neurol. Neurosurg. Psych. 42: 368–372, 1979.Google Scholar
  43. [43]
    Stibler H.:The normal cerebrospinal fluid proteins identified by means of thin-layer isoelectric focusing and crossed immunoelectrofocusing. J. Neurol. Sci., 36:273–288, 1978.CrossRefPubMedGoogle Scholar
  44. [44]
    Chiodi F., Siden A., Osby E.:Isoelectric focusing of monoclonal immunoglobulins G. A and M followed by detection with the avidin-biotin system. Electrophoresis, 6:124–128, 1985.CrossRefGoogle Scholar
  45. [45]
    Henriksson A.M., Kam-Hansen S., Andersson R. Immunoglobulin-producing cells in CSF and blood from patients with multiple sclerosis and other inflammatory neurological diseases enumerated by protein-A plaque assay. J. Neuroimmunol., 1:229–302, 1981.CrossRefGoogle Scholar
  46. [46]
    Santoli D., Defreitas E.C. Sandberg-Wollhem M., Francis M.K., Koprowski H.:Phenotipic and functional characterization of T cell clones derived from the CSF of multiple sclerosis patients. J. Immunol. 132:2386–2392, 1984.PubMedGoogle Scholar

Copyright information

© Masson Italia Editori 1987

Authors and Affiliations

  • Laurenzi M. A. 
    • 1
  • Provinciali L. 
    • 1
  • Giovagnoli A. R. 
    • 1
  • Busco
    • 2
  • Bocchini V. 
    • 3
  • Paris L. 
    • 4
  • Chiodi F. 
    • 4
  1. 1.Istituto Malattie del Sistema NervosoUniversità di AnconaItaly
  2. 2.Laboratorio AnalisiUSL n. 12AnconaItaly
  3. 3.Istituto di Biologia GeneraleUniversità di PerugiaItaly
  4. 4.Clinica Neurologica IIIUniversità di RomaItaly

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