Naïve mice were immunized with 50μg of PLP peptide (either P178 or P139) or OVA323–339 in CFA. At day 14, spleens and inguinal lymph nodes were collected and reactivated in culture with cognate antigens and hIL-2 for 72hr as previously described (4 (link), 7 (link)–11 (link), 18 ). At the end of culture, cells were collected, and CD8 T cells were magnetically isolated using Ly-2 microbeads (Miltenyi) and 10×106 live cells were transferred to recipient mice. Mice received CD8 T cell transfers i.v. prior to disease induction (d-1) or during the acute phase of disease (d11).
Protocol detail
Adoptive Transfer of Antigen-Specific CD8 T Cells
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Corresponding Organization :
Other organizations : University of Iowa Health Care, University of Iowa, Veterans Health Administration
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Variable analysis
independent variables
- Immunization with 50μg of PLP peptide (either P178 or P139)
- Immunization with 50μg of OVA323–339 in CFA
- Timing of CD8 T cell transfers (d-1 or d11)
- CD8 T cell response (as measured by cell transfer experiments)
- CFA in immunization
- Culture conditions (cognate antigens and hIL-2 for 72hr)
- Cell isolation method (magnetic isolation using Ly-2 microbeads)
- Number of live cells transferred (10×10^6)
- Positive control: OVA323–339 immunization
- Negative control: Not explicitly mentioned
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