Figure 1. CSF T cell recognition and confirmation assay.

PBMCs were obtained from apheresis samples or blood, and monocytes were isolated via positive selection with CD14⁺ magnetic beads. For the primary proliferation assay, monocytes were differentiated into immature dendritic cells (iDC). On the sixth day of culture, iDCs were loaded with selected Ag's and stimulation cocktail was added to induce DC maturation. 48 hours later, mature DCs (mDC) were co-cultured with peripheral and intrathecal T cells. After seven days of proliferation, cultured T cells were re-stimulated overnight with newly differentiated, identically loaded mDCs. Ag-specificity was detected by flow cytometric analysis of cytokine-producing CD4⁺ and CD8⁺ T cells.