Characterization of proteolipid protein-peptide-specific CD(4)(+) T cell of experimental allergic encephalomyelitis in Biozzi AB/H mice.
OBJECTIVE
To detect the function of proteolipid protein (PLP) peptide (residues 56 - 70)-specific CD(4)(+) T cells in experimental allergic encephalomyelitis (EAE) in Biozzi AB/H mice (H-2A(g7)).


METHODS
Biozzi AB/H mice were immunized by synthetic PLP(56 - 70) peptide (DYEYLINVIHAFQYV) which was emulsified by sonication with complete Freund's adjuvant, a EAE model proven histologically and clinically. Murine splenocytes and spinal cord infiltrated (SCI) T cells were stimulated by PLP(56 - 70), then the CD(4)(+) T cells were isolated by Dynabeads, and confirmed by staining with anti-CD(4) antibody. Finally, the IL2 bioassay and IFN-gamma/IL4 ELISA were done to detect T cell proliferation and cytokine secretion after PLP(56 - 70) stimulation.


RESULTS
The histology of murine spinal cord showed a great number of lymphocytes infiltrated the spinal cord; the clinical signs showed high scores (4.3) on the peak, as well as a good EAE model. After being isolated by Dynabeads, CD(4)(+) T cells showed high purification (> 99%) by staining with anti-CD(4) antibody. IL2 bioassay showed that those T cells were PLP(56 - 70)-specific T cells. ELISA showed that those T cells had high IFN-gamma/IL4 ratio, indicating that they are T helper 1 (Th1) cells.


CONCLUSIONS
PLP(56 - 70)-specific splenocytes and SCI CD(4)(+) T cells in EAE from Biozzi AB/H mice were detected and showed that both of them were PLP(56 - 70)-specific Th1 cells. It is beneficial to understand what kind of role these T cells play in the development of EAE.
Department of Neurology, The Third Affiliated Hospital, Hunan Medical University, Changsha 410013, China. yongpeng70@yahoo.com
Yong
Y
Peng
Chih-pin
CP
Liu
D000595:Amino Acid Sequence; D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000276:immunology* / Q000473:pathology; D007371:Interferon-gamma / Q000378:metabolism; D007376:Interleukin-2 / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D018991:Myelin Proteolipid Protein / Q000737:chemistry / Q000276:immunology*; D010446:Peptide Fragments / Q000008:administration & dosage / Q000276:immunology*; D013154:Spleen / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D018417:Th1 Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism
D007376:Interleukin-2; D018991:Myelin Proteolipid Protein; D010446:Peptide Fragments; D015847:Interleukin-4; D007371:Interferon-gamma