Yong Peng1, Chih-pin Liu. 1. Department of Neurology, The Third Affiliated Hospital, Hunan Medical University, Changsha 410013, China. yongpeng70@yahoo.com
Abstract
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.
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.