Skip to main content
PMC home page
Elsevier - PMC COVID-19 Collection logoLink to Elsevier - PMC COVID-19 Collection
. 2002 Dec 9;4(2):129–140. doi: 10.1016/0165-5728(83)90017-6

Improvements in obtaining and characterizing mouse cerebrospinal fluid☆

Application to mouse hepatitis virus-induced encephalomyelitis

John O Fleming 1,∗∗, Jenny YP Ting 2,∗, Stephen A Stohlman 1,2, Leslie P Weiner 1,2
PMCID: PMC7172882  PMID: 6300186

Abstract

This report describes advances in techniques for analyzing cellular and humoral immune components in the cerebrospinal fluid (CSF) of the mouse that are applicable to other laboratory animals. CSF studies undertaken during experimental infection of mice with JHM strain virus (JHMV) of mouse hepatitis virus are presented. A critical pitfall which can lead to erroneous or invalid results is contamination of the CSF by even minute quantities of blood. Means of avoiding this contamination are attention to anatomical reference points, the use of a micropipet, and prior intracardiac perfusion of animals with phosphate-buffered saline. Cells in the CSF were typed as either B, T, polymorphonuclear, or mononuclear cells by the combination of a microcytotoxicity assay and histologic stains. A radioimmunoassay (RIA) allowed quantification of antibodies to JHMV in the CSF and indicated the presence of intrathecal synthesis of antibody in chronically infected mice. The combined use of these sensitive methods makes possible CSF analysis in individual mice rather than in pooled groups.

Keywords: Cerebrospinal fluid, Encephalomyelitis, JHM strain virus, Mouse hepatitis virus

Footnotes

☆

This work is supported in part by grants NS-7149 and NS-18146 awarded by the NIH. JOF is a recipient of postdoctoral fellowship grant FG-544-A-1 from the National Multiple Sclerosis Society.

References

  1. Carp R.I., Davidson A.I., Merz P.A. A method for obtaining cerebrospinal fluid from mice. Res. Vet. Sci. 1971;120:449. [PubMed] [Google Scholar]
  2. Cheever F.S., Daniels J.B., Pappenheimer A.M., Bailey O.T. A murine virus (JHM) causing disseminated encephalomyelitis with extensive destruction of myelin. J. Exp. Med. 1949;90:181–194. doi: 10.1084/jem.90.3.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cutler R.W.P. Neurochemical aspects of blood-brain-cerebrospinal fluid barriers. In: Wood J.H., editor. Neurobiology of Cerebrospinal Fluid. Plenum Press; New York, NY: 1980. p. 41. [Google Scholar]
  4. Dawson H. Churchill; London: 1967. Physiology of the Cerebrospinal Fluids. [Google Scholar]
  5. Doherty P.C., Gerhard W. Breakdown of the blood-cerebrospinal fluid barrier to immunoglobulin in mice injected intracerebrally with a neurotropic influenza A virus. J. Neuroimmunol. 1981;1:227–237. doi: 10.1016/0165-5728(81)90027-8. [DOI] [PubMed] [Google Scholar]
  6. Fenstermacher J.D., Patlak C.S. The exchange of material between cerebrospinal fluid and brain. In: Cserr H.F., Fenstermacher J.D., Fencl V., editors. Fluid Environment of the Brain. Academic Press; New York, NY: 1975. pp. 201–214. [Google Scholar]
  7. Fishman R.A. W.B. Saunders; Philadelphia, PA: 1980. Cerebrospinal Fluid in Diseases of the Nervous System. [Google Scholar]
  8. Frelinger J.A., Niederhuber J.E., David C.S., Shreffler D.C. Evidence for the expression of Ia (H-2 associated) antigens on thymus-derived lymphocytes. J. Exp. Med. 1974;140:1273–1284. doi: 10.1084/jem.140.5.1273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gerhard W., Iwasaki Y., Koprowski H. The central nervous system-associated immune response to parainfluenza type 1 virus in mice. J. Immunol. 1978;120:1256–1260. [PubMed] [Google Scholar]
  10. Greaves M.F., Owen J.J.T., Raff M.C. Elsevier; Amsterdam: 1974. T. and B lymphocytes — Origins, properties, and roles in immune responses; p. 77. [Google Scholar]
  11. Griffin D.E. Immunoglobulins in the cerebrospinal fluid — Changes during acute viral encephalitis in mice. J. Immunol. 1981;126:27–31. [PubMed] [Google Scholar]
  12. Lipton H.L., Gonzales-Scarano F. Central nervous system immunity in mice infected with Theiler's virus, Part I (Localizing neutralizing antibody response) J. Infect. Dis. 1978;137:145–151. doi: 10.1093/infdis/137.2.145. [DOI] [PubMed] [Google Scholar]
  13. Lisak R.P., Zweimann B. In vitro cell-mediated immunity of cerebrospinal lymphocytes to myelin basic protein in primary demyelinating diseases. N. Engl. J. Med. 1977;297:850–853. doi: 10.1056/NEJM197710202971602. [DOI] [PubMed] [Google Scholar]
  14. Medawar P.B. Immunity to homologous grafted skin, Part 3 (Fate of skin homographs transplanted to the brain, to subcutaneous tissue, and to the anterior chamber of the eye. Brit. J. Exp. Path. 1948;299:58–69. [PMC free article] [PubMed] [Google Scholar]
  15. Moxon R.E., Ostrow P.T. Haemophilus influenzae meningitis in infant rats — Role of bacteremia in pathogenesis of age-dependent inflammatory responses in cerebrospinal fluid. J. Infect. Dis. 1977;135:303–307. doi: 10.1093/infdis/135.2.303. [DOI] [PubMed] [Google Scholar]
  16. Nathanson N., Petursson G., Georgsson G., Palsson P.A., Martin J.R., Miller A. Pathogenesis of visna, Part 4 (Spinal fluid studies) J. Neuropath. Exp. Neurol. 1979;38:197–208. doi: 10.1097/00005072-197905000-00001. [DOI] [PubMed] [Google Scholar]
  17. Robb J.A., Bond C.W. Pathogenic murine coronaviruses. Virology. 1979;94:352–370. doi: 10.1016/0042-6822(79)90467-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rosenthal J.D., Hayashi K., Notkins A.L. Comparison of direct and indirect solid-phase microradio-immunoassays for the detection of viral antigens and antiviral antibody. Appl. Microbiol. 1973;25:567. doi: 10.1128/am.25.4.567-573.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Russell E.S., Neufeld E.F., Higgins C.T. Vol. 78. 1951. Comparison of normal blood picture of young adults from 18 in bred strains of mice; pp. 761–766. (Proc. Soc. Exp. Biol. Med.). [DOI] [PubMed] [Google Scholar]
  20. Stohlman S.A., Weiner L.P. Chronic central nervous system demyelination in mice after JHM virus infection. Neurology (Minneap.) 1981;31:38–44. doi: 10.1212/wnl.31.1.38. [DOI] [PubMed] [Google Scholar]
  21. Stohlman S.A., Frelinger J.A., Weiner L.P. Resistance to fatal central nervous system disease by mouse hepatitis virus, strain JHM, Part 2 (Adherent cell mediated protection) J. Immunol. 1980;124:1733–1739. [PubMed] [Google Scholar]
  22. Stohlman S.A., Brayton P.R., Lai M.C., Fleming J.O., Weiner L.P. Murine coronaviruses — Isolation and characterization of two plaque morphology variants of a neurotropic strain JHM. J. Gen. Virol. 1982;63:265–275. doi: 10.1099/0022-1317-63-2-265. [DOI] [PubMed] [Google Scholar]
  23. Tourtellotre W. On cerebrospinal fluid immunoglobulin-G (IgG) quotients in multiple sclerosis and other disease. J. Neurol. Sci. 1970;10:279–304. doi: 10.1016/0022-510x(70)90156-5. [DOI] [PubMed] [Google Scholar]
  24. Trotter J.L., Brooks B.R. Pathophysiology of cerebrospinal fluid immunoglobulins. In: Wood J.H., editor. Neurobiology of Cerebrospinal Fluid. Plenum Press; New York, NY: 1980. pp. 465–486. [Google Scholar]
  25. Von Boehmer H., Sprent J., Nabholz M. Tolerance to histocompatibility determinants in tetraparental bone marrow chimeras. J. Exp. Med. 1975;141:322–334. doi: 10.1084/jem.141.2.322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Weiner L.P. Pathogenesis of demyelination induced by a mouse hepatitis virus (JHM virus) Arch. Neurol. (Chic.) 1973;28:298–303. doi: 10.1001/archneur.1973.00490230034003. [DOI] [PubMed] [Google Scholar]
  27. Wertlake P.T., Marokovitz B.A., Stellar S. Cytologic evaluation of cerebrospinal fluid with clinical and histologic correlation. Acta Cytol. 1972;16:224–239. [PubMed] [Google Scholar]
  28. Williams P.L., Warwick R. Churchill Livingstone; London: 1975. Functional Neuroanatomy of Man; p. 858. [Google Scholar]
  29. Yam L.T., Li C.Y., Crosby W.H. Cytochemical identification of monocytes and granulocytes. Amer. J. Clin. Path. 1971;55:283–290. doi: 10.1093/ajcp/55.3.283. [DOI] [PubMed] [Google Scholar]
  30. Zinkernagel R.M., Doherty P.C. Cytotoxic thymus-derived lymphocytes in cerebrospinal fluid of mice with lymphocytic choriomeningitis. J. Exp. Med. 1973;138:1266–1269. doi: 10.1084/jem.138.5.1266. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Neuroimmunology are provided here courtesy of Elsevier

RESOURCES