Journal of Biomedical Science

, Volume 4, Issue 6, pp 300–307 | Cite as

Infiltrated cells in experimental allergic encephalomyelitis by additional intracerebral injection in myelin-basic-protein-sensitized B6 mice

  • Tzong-Shiann Ho
  • Chia-Ying Tsai
  • Nina Tsao
  • Nan-Haw Chow
  • Huan-Yao Lei
Original Paper


We previously reported that murine experimental allergic encephalomyelitis can be induced by an additional intraperitoneal and intracerebral (i.c.) restimulation in resistant B6 mice after standard immunization with myelin antigens in complete Freund's adjuvant andBordetella pertussis coadjuvant. Neutrophils infiltrated into perivascular spaces at 12 h, followed by mononuclear cells 24 h after i.c. injection. In this study, we report that the i.c. injection induced the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). The kinetic expression of ICAM-1 or VCAM-1 on brain endothelial cells paralleled the infiltration of neutrophils and mononuclear cells, respectively. The infiltrated lymphocytes also expressed very late antigen-4 (VLA-4) molecules. The microvascular endothelial cells were positive for VCAM-1, whereas the surrounding mononuclear cells were VLA-4 positive. Furthermore, we found a unique subpopulation of cells with characteristics of CD4-CD8-Vβ8+ markers. The kinetic studies of this population showed that these cells were transiently depleted from 12 to 24 h after i.c. challenge (before the development of clinical symptoms) in cervical lymph nodes. These CD4-CD8-Vβ8+ cells can be expanded by in vitro culture with myelin basic protein or IL-2. No significant changes of CD4+/CD8+ cells were noted. CD4+CD8CD3+ cells were also found in brain by double histochemical stains and were the major infiltrating cells at 24 or 48 h after i.c. challenge.

Key Words

Experimental allergic encephalomyelitis Adhesion molecule CD4CD8CD3+ cells Intracerebral stimulation Cervical lymphatics 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Acha-Orbea H, Steinman L, McDevitt HO. T cell receptors in murine autoimmune diseases. Ann Rev Immunol 7:371–405;1989.Google Scholar
  2. 2.
    Baron JL, Madri JA, Ruddle NH, Hashim G, Janeway CA. Surface expression of α4 integrin by CD4 T cells is required for their entry into brain parenchyma. J Exp Med 177:56–78;1993.Google Scholar
  3. 3.
    Bendelac A, Killeen N, Littman DR, Schwartz RH. A subset of CD4+ thymocytes selected by MHC class I molecules. Science 263:1774–1778;1994.Google Scholar
  4. 4.
    Bradbury MW, Cserr HF. Drainage of cerebrospinal fluid and brain interstitial fluid into cervical lymphatics. In: Johnson MG, eds. Experimental Biology of the Lymphatic Circulation. New York, Elsevier, 355–394;1985.Google Scholar
  5. 5.
    Brown A, McFarlin DE, Raine CS. Chronologic neuropathology of relapsing experimental allergic encephalomyelitis in the mouse. Lab Invest 46:171–185;1982.Google Scholar
  6. 6.
    Chou FCH, Chou CHJ, Shapira R, Kibler RF. Basis of microheterogeneity of MBP. J Biol Chem 251:2671–2679;1976.Google Scholar
  7. 7.
    Claudio L, Raine CS, Brosnan CF. Evidence of persistent blood-brain barrier abnormalities in chronic-progressive multiple sclerosis. Acta Neuropathol 90:228–238;1995.Google Scholar
  8. 8.
    Crispe IN. CD4/CD8-negative T cells with αβ antigen receptors. Curr Opin Immunol 6:438–441;1994.Google Scholar
  9. 9.
    Cserr HF, Knopf PM. Cervical lymphatics, the blood-brain barrier and the immunoreactivity of the brain: A new view. Immunol Today 13:507–512;1992.Google Scholar
  10. 10.
    Cua JD, Hinton DR, Stohlman SA. Self-antigen-induced Th2 responses in experimental allergic encephalomyelitis (EAE)-resistant mice. Th2-mediated suppression of autoimmune disease. J Immunol 155:4052–4059;1995.Google Scholar
  11. 11.
    Erard F, Wild MT, Garcia-Sanz JA, Le Gros G. Switch of CD8 T cells to noncytolytic CD4CD8 cells that make Th2 cytokines and help B cells. Science 260:1802–1805;1993.Google Scholar
  12. 12.
    Gordon LB, Kahn M, Cserr HF, Knopf PM. Comparison of serum antibody responses to CNS and systemically administered ovalbumin (OVA). Soc Neurosci Abstr 16:1209;1990.Google Scholar
  13. 13.
    Hafler DA, Weiner HL. MS: A CNS and systemic autoimmune disease. Immunol Today 10:104–107;1989.Google Scholar
  14. 14.
    Harling-Berg CJ, Knopf PM, Merian J, Cserr HF. Role of cervical lymph nodes in the systemic humoral response to human serum albumin microinfused into rat CSF. J Neuroimmunol 25:185–193;1989.Google Scholar
  15. 15.
    Harris JO, Frank JA, Patronas N, McFarlin DE, McFarland HF. Serial gadolinium-enhanced magnetic resonance imaging scans in patients with early, relapsing-remitting multiple sclerosis: Implications for clinical trials and natural history. Ann Neurol 29:548–555;1991.Google Scholar
  16. 16.
    Johnson RT, Hirsch RL, Griffin DE, Wolinsky JS, Roedenbeck S, de Soriano IL, Vaisberg A. Clinical and immunological studies of measles encephalitis. Trans Am Neurol Assoc 106:42–45;1981.Google Scholar
  17. 17.
    Liebert UG, Linington C, Meulen V. Induction of autoimmune reactions to myelin basic protein in measles virus encephalitis in Lewis rats. J Neuroimmunol 17:103–118;1988.Google Scholar
  18. 18.
    Matsuda M, Tsukada N, Miyagi K, Yanagisawa N. Adhesion of lymphocytes to endothelial cells in experimental allergic encephalomyelitis before and after treatment with endotoxin lipopolysaccharide. Int Arch Allergy Immunol 106:335–344;1995.Google Scholar
  19. 19.
    Mokhtarian F, McFarlin DE, Raine CS. Adoptive transfer of myelin basic protein-sensitized T cells produces chronic relapsing demyelinating disease in mice. Nature 309:356–358;1984.Google Scholar
  20. 20.
    Mukherjee A, Vogt RF, Linthicum DS. Measurement of myelin basic protein by radioimmunoassay in closed head trauma, multiple sclerosis and other neurological diseases. Clin Biochem 18:304–307;1985.Google Scholar
  21. 21.
    Paterson PY, Day ED. Current perspectives of neuroimmunologic disease: Multiple sclerosis and experimental allergic encephalomyelitis. Clin Immunol Rev 1:581–697;1981–1982.Google Scholar
  22. 22.
    Peterson PY, Swanborg RH. Demyelinating diseases of the central and peripheral nervous system. In: Samter M, ed. Immunological Disease, ed 4. Boston, Little Brown, 1877–1916;1988.Google Scholar
  23. 23.
    Pettinelli CB, McFarlin DE. Adoptive transfer of experimental allergic encephalomyelitis in SJL/J mice after in vitro activation of lymph node cells by myelin basic protein: Requirement of Lyt 1+2- T lymphocytes. J Immunol 127:1420–1423;1981.Google Scholar
  24. 24.
    Raine CS. Biology of disease. Analysis of autoimmune demyelination: Its impact upon multiple sclerosis. Lab Invest 50:608–635;1984.Google Scholar
  25. 25.
    Reimann J. Double-negative (CD4CD8), TCRαβ-expressing, peripheral T cells. Scand J Immunol 34:679–688;1991.Google Scholar
  26. 26.
    Reimann J, Bellan A, Conrad P. Development of autoreactive L3T4+ T cells from double-negative (L3T4/Ly-2) Thy-1+ spleen cells of normal mice. Eur J Immunol 18:989–999;1988.Google Scholar
  27. 27.
    Seman M, Boudaly S, Roger T, Morisset J, Pham G. Autoreactive T cells in normal mice: Unrestricted recognition of self peptides on dendritic cell I-A molecules by CD4CD8 T cell receptor α/β+ T cell clones expressing Vβ8.1 gene segments. Eur J Immunol 20:1265–1272;1990.Google Scholar
  28. 28.
    Shaw MK, Kim C, Ho KL, Lisak RP, Tse HY. A combination of adoptive transfer and antigenic challenge induces consistent murine experimental autoimmune encephalomyelitis in C57BL/6 mice and other reputed resistant strains. J Neuroimmunol 39:139–150;1992.Google Scholar
  29. 29.
    Steffen BJ, Butcher EC, Engelhardt B. Evidence for involvement of ICAM-1 and VCAM-1 in lymphocyte interaction with endothelium in experimental autoimmune encephalomyelitis in the central nervous system in SJL/J mouse. Am J Pathol 145:189–201;1994.Google Scholar
  30. 30.
    Swanborg RH. Experimental allergic encephalomyelitis. Methods Enzymol 162:413–421;1988.Google Scholar
  31. 31.
    de Talance A, Regnier D, Spinelli S, Morisset J, Seman M. Origin of autoreactive T helper cells. I. Characterization of Thy-1+, Lyt, L3T4 precursors in the spleen of normal mice. J Immunol 137:1101–1108;1986.Google Scholar
  32. 32.
    Traugott U. Detailed analysis of early immunopathologic events during lesion formation in acute experimental autoimmune encephalomyelitis. Cell Immunol 119:114–129;1989.Google Scholar
  33. 33.
    Traugott U, Raine CS, McFarlin DE. Acute experimental allergic encephalomyelitis in the mouse: Immunopathology of the developing lesion. Cell Immunol 91:240–254;1985.Google Scholar
  34. 34.
    Tsai CY, Chow NH, Ho TS, Lei HY. Intracerebral injection of myelin basic protein (MBP) induces inflammation in brain and causes paraplegia in MBP-sensitized B6 mice. Clin Exp Immunol 109:(in press)1997.Google Scholar
  35. 35.
    Tuohy VK, Sobel RA, Lees MB. Myelin proteolipid protein-induced experimental allergic encephalomyelitis. Variation of disease expression in different strains of mice. J Immunol 140:1868–1873;1988.Google Scholar
  36. 36.
    Waksman BH, Reynolds WE. Multiple sclerosis as a disease of immune regulation. Proc Soc Exp Biol Med 175:282–294;1984.Google Scholar
  37. 37.
    Wang SD, Huang KJ, Lin YS, Lei HY. Sepsis-induced apoptosis of the thymocyte in mice. J Immunol 152:5014–5021;1994.Google Scholar
  38. 38.
    Wilcox CE, Ward AMV, Evans A, Baker D, Rothlein R, Turk JL. Endothelial cell expression of the intercellular adhesion molecule-1 (ICAM-1) in the central nervous system of guinea pig during acute and chronic relapsing experimental allergic encephalomyelitis. J Neuroimmunol 30:43–51;1990.Google Scholar
  39. 39.
    Yamada S, DePasquale M, Patlak SC, Cserr HF. Albumin outflow into deep cervical lymph from different regions of rabbit brain. Am J Physiol 261:H1197-H1204;1991.Google Scholar

Copyright information

© National Science Council 1997

Authors and Affiliations

  • Tzong-Shiann Ho
    • 1
  • Chia-Ying Tsai
    • 1
  • Nina Tsao
    • 1
  • Nan-Haw Chow
    • 2
  • Huan-Yao Lei
    • 1
  1. 1.Department of Microbiology and ImmunologyCollege of Medicine, National Cheng Kung UniversityTainanTaiwan(ROC)
  2. 2.Department of PathologyCollege of Medicine, National Cheng Kung UniversityTainanTaiwan

Personalised recommendations