Stimulation of class lrestricted responses by BMM0

Equipment and reagents

CD8 * class l-restricted T cell hybridoma Specific and irrelevant soluble antigens, e.g. ovalbumin, hen egg lysozyme (Sigma)

Specific peptides corresponding to know class l-restricted epitopes of the chosen antigens

Method

1 Plate antigen loaded BMM0 (as in Protocol 5) or untreated BMM0 into 96-well plates as in Protocol 3, steps 2-5.

2 Pulse untreated macrophages with soluble antigen or peptides as described in Protocol 3, steps 6-9/

3 Assay IL-2 production from CD8+ T cell hybridomas, or CTL activity of primed T cells against peptide pulsed or transfected target cells, as described above.

a The dose range for soluble antigens should extend up to 1 mg/ml. Peptides (recognized by the responding T cells) should be tested over 3-4 log range of concentration.

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