Characterization of proteolipid protein-peptide-specific CD_4~+ T cell of experimental allergic ence

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To detect the function of proteolipid protein (PLP) peptide (residues 56-70) sp ecific CD 4 + T cells in experimental allergic encephalomyelitis (EAE) in Bioz zi AB/H mice (H 2A g7 ) Methods Biozzi AB/H mice were immunized by synthetic PLP 56 70 peptide (DYEYLINVIH AFQYV) 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 γ/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 infiltr ated 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 Dynabea ds , 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 γ/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 Bioz zi AB/H mice were detected and showed that both of them were P LP 56 70 specific Th1 cells It is beneficial to understand what kind o f role these T cells play in the development of EAE To detect the function of proteolipid protein (residues 56-70) sp ecific CD 4 + T cells in experimental allergic encephalomyelitis (EAE) in Bioz zi AB / H mice (H 2A g7) Methods Biozzi AB / H mice were immunized by synthetic PLP 56 70 peptide (DYEYLINVIH AFQYV) 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 γ / 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 infiltr ated 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 Dy CD4 + T cells showed high purification (> 99%) by staining with anti CD 4 antibody IL2 bioassay showed those T cells were PLP 56 70 specific T cells ELISA showed those those T cells had high IFN γ / 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 Bioz zi AB / H mice were detected and showed that both of them were P LP 56 70 specific Th1 cells It is beneficial to understand what kind of role these T cells play in the development of EAE
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