Table 4.

Identification of the COA-1-derived epitopes recognized by the CD4⁺ clone C111

Stimulator	HLA-DRβ1	No peptide
None		<8a
1869 col	0402, 1301	2186
1681 CD40LB	0301, 0402	<8
1847 CD40LB	0401, 1301	<8
			Peptide concentration (μg/mM)
		Peptideb	100	50	25	12.5	6.25
1681 CD40LB	0301, 0402	FSTFPPTLYQDDTLTLQAAG	105	236	69	<7.8	<7.8
1681 CD40LB		TLYQDDTLTLQAAGLVPKAA	51	159	<7.8	<7.8	<7.8
1681 CD40LB		DDTLTLQAAGLVPKAALLLRA	11	16	<7.8	<7.8	<7.8
1681 CD40LB		LQAAGLVPKAALLLRARRAP	21	12	<7.8	<7.8	<7.8
1847 CD40LB	0401, 1301	ASAFEIFSTFPPTLYQDDTL	<	<7.8	<7.8	<7.8	<7.8
1847 CD40LB		FSTFPPTLYQDDTLTLQAAG	226	397	296	79	<7.8
1847 CD40LB		TLYQDDTLTLQAAGLVPKAA	79	326	<7.8	<7.8	<7.8
1847 CD40LB		DDTLTLQAAGLVPKAALLLRA	22	33	<7.8	<7.8	<7.8
1847 CD40LB		LQAAGLVPKAALLLRARRAP	52	32	<7.8	<7.8	<7.8

^aThe CD4⁺ T-cell clone C111 was added at 2 × 10⁴ cells/well at the final volume of 250 μl/well of ISCOVE’s plus 10% HS, and after 18 h of incubation, the supernatants were collected and the IFN-γ release was evaluated by ELISA.

^bPeptides of 20 or 21 amino acids overlapping by 15 amino acids were synthesized using the putative COA-1 protein, in the 1D8 region (1012–1318 bp). 4 × 10⁵/ml of B cells sharing one of the DRβ1 molecules (*0402 or *1301) with the autologous tumor 1869, were incubated for 3 h at 37°C and 5% CO₂ in the presence or not (−) of the peptides at the final volume of 100 μl/well in ISCOVE’s plus 10% HS.