Table 2.

Identification of target peptide-specific T cell clones from tetramer^pos CD8^pos sorted T cells using healthy donor PBMCs

Donora	T cells sorted (n)b	Clones exp. (n)c	Reactivity of T cell clonesd				Peptide specificity of T cell clonese
			Not reactive (n)	Off-target (n)	HLA-A1/A24 (n)	HLA-B8/B35 (n)	p236 BOB1 B35	p243 FCRL5 A1	p247 TLR10 B8	p248 MIST A1	p252 FCRL2 B8	p256 TLR10 B8	p259 TNFRSF13B B35	p261 RALGPS2 B8	p263 FCRL5 A1	p265 TLR10 A1	p269 VPREB3 A24
3	288	239	233	6	0	0
4	192	156	101	54	1	0		1x
5	192	129	44	83	0	2							2x
6	528	433	235	180	6	12	6x		1x	2x	3x		1x	1x		3x	1x
7	288	207	181	26	0	0
8	384	311	289	22	0	0
12	864	639	320	314	5	0		1x		2x					2x
13	1,728	1,002	891	108	2	1						1x					2x
14	1,296	384	330	53	0	1						1x
15	864	449	399	49	0	1								1x
16	1,920	1,018	907	105	3	3				1x			2x	1x			2x
17	2,640	360	351	9	0	0
18	1,152	192	143	40	6	3				2x	2x	1x				2x	2x
Total	12,336	5,519	4,424	1,049	23	23	6x	2x	1x	7x	5x	3x	5x	3x	2x	5x	7x

^aArbitrary donor numbers assigned to healthy donors from which PBMCs were obtained, in accordance with Table S3.

^bNumber of pHLA-tetramer^pos CD8^pos T cells single-cell sorted from each donor.

^cNumber of single-cell-sorted T cells that expanded and were tested for peptide specificity.

^dExpanded T cell clones were stimulated with a 1:1 mixture of HLA-A1 and HLA-A24 Td K562 cells or HLA-B8 and B35 Td K562 cells, unloaded or loaded with a mixture of all target peptides (100 nM). Reactivity was assessed after overnight co-culture based on IFN-γ ELISA; for T cell clones not producing IFN-γ, GM-CSF was used instead. Recognition was categorized as: not reactive, off-target recognition, HLA-A1/A24-restricted peptide-specific recognition (HLA-A1/A24), and HLA-B8/B35-restricted peptide-specific recognition (HLA-B8/B35).

^eThe number of peptide-specific T cell clones per donor is indicated. Specificity was determined as demonstrated in Figure 2. No T cell clones were identified for peptides not included in the table.