Table 3.

HLA-A*0201–Restricted Cytokine Release by PBMC Cultures Sensitized in Vitro with MART-1_27–35

	IFN-γ release (pg/5 × 105 effectors/24 hours)
	A		C
Patient #	T2+MART-1	T2+Flu M1	T2+MART-1	T2+Flu M1
1	476	229	7839	273
2	10	0	360	0
3	498	460	6785	322
4	1614	88	8580	0
5	5976	310	27,080	0
6	262	0	25,620	0
7	131	83	5952	0
8	0	0	7499	0
9	12	18	960	436
10	696	68	23,760	80
11	1146	893	3240	0
12	82	0	1570	0
13	1664	44	2668	0
14	46	0	192	0
15	540	0	4336	0
16	0	0	64	0
17	1864	956	3264	56
18	60	0	8848	0

Release of cytokine is expressed as the amount of IFN-γ (pg/mL) secreted by 5 × 10⁵ effector cells/mL CTL cocultured for 24 hours in the presence of 5 × 10⁵ relevant (T2 + MART-1_27–35) or irrelevant (T2 + Flu M1_58–66) stimulators per milliliter. The assay was performed after three in vitro stimulations.

Specific release of IFN-γ by a PBMC culture was defined as (1) threefold or higher difference in IFN-γ production in the presence of relevant (T2 + MART-1_27–35) vs irrelevant (T2 + Flu M1_58–66) stimulation and (2) ≥ 100 pg/10⁶ cells/24 hours production of IFN-γ. Difference in CTL reactivity was considered a threefold increase in specific release between prevaccination and postvaccination cultures.

Abbreviations: A = preimmunization CTL culture; C = postimmunization PBMC.