TABLE 4.

Recognition of DCs Transfected with gp100 cDNA in the Presence or Absence of Maturation Cytokines by HLA-A^*0201 (A2), HLA-DRβ1^*0401 (DR4), and HLA-DRβ1^*0701 (DR7) Restricted, gp100-reactive T-cell Populations^*

Target Cells	HLA Expression	Preloaded Peptide^†	Genetic Modification	Maturation Cytokines^‡	CK3H6 (A2) Exp. 1	JR1E2 (A2) Exp. 1	B104 (DR7) Exp. 1	CL70 (A2) Exp. 2	L2D8 (A2) Exp. 2	BR-B8 (DR4) Exp. 2
Donor 5 DC	A2+ DR7+	—	gp100 cDNA	None	72	76	81	—	—	—
Donor 5 DC	A2+ DR7+	—	GFP cDNA	None	104	71	834^§	—	—	—
Donor 5 DC	A2+ DR7+	—	gp100 cDNA	TNFα + PGE₂	75	67	76	—	—	—
Donor 5 DC	A2+ DR7+	—	GFP cDNA	TNFα + PGE₂	76	76	71	—	—	—
Donor 6 DC	A2+ DR7+	—	gp100 cDNA	None	72	71	78	—	—	—
Donor 6 DC	A2+ DR7+	—	GFP cDNA	None	212	69	> 2000	—	—	—
Donor 6 DC	A2+ DR7+	—	gp100 cDNA	TNFα + PGE₂	67	68	70	—	—	—
Donor 6 DC	A2+ DR7+	—	GFP cDNA	TNFα + PGE₂	78	68	77	—	—	—
Donor 7 DC	A2⁺DR4⁺	—	gp100 cDNA	None	—	—	—	20	84	4605
Donor 7 DC	A2⁺DR4⁺	—	GFP cDNA	None	—	—	—	88	88	250
Donor 7 DC	A2⁺DR4⁺	—	gp100 cDNA	TNFα + PGE₂	—	—	—	19	58	534
Donor 7 DC	A2⁺DR4⁺	—	GFP cDNA	TNFα + PGE₂	—	—	—	130	98	238
Donor 7 DC	A2⁺DR4⁺	—	gp100 cDNA	CD40L+LPS	—	—	—	205	1285	3527
Donor 7 DC	A2⁺DR4⁺	—	GFP cDNA	CD40L+LPS	—	—	—	251	390	524
Donor 8 DC	A2⁺DR4⁺	—	gp100 cDNA	None	—	—	—	22	263	11737
Donor 8 DC	A2⁺DR4⁺	—	GFP cDNA	None	—	—	—	33	162	341
Donor 8 DC	A2⁺DR4⁺	—	gp100 cDNA	TNFα + PGE₂	—	—	—	22	246	2324
Donor 8 DC	A2⁺DR4⁺	—	GFP cDNA	TNFα + PGE₂	—	—	—	69	158	313
Donor 8 DC	A2⁺DR4⁺	—	gp100 cDNA	CD40L+LPS	—	—	—	54	603	4973
Donor 8 DC	A2⁺DR4⁺	—	GFP cDNA	CD40L+LPS	—	—	—	407	309	381
T2 or EBV-B	T cell matched	control	—	—	65	65	70	18	22	144
T2 or EBV-B	T cell matched	relevant	—	—	> 2000	> 2000	> 2000	3981	11662	6714
F002Rmel CIITA	A2⁺DR4⁻DR7⁺	—	VSV-GFP	—	70	76	93	—	—	—
F002Rmel CIITA	A2⁺DR4⁻DR7⁺	—	VSV-gp100	—	1676	> 2000	> 2000	—	—	—
526mel CIITA	A2⁺DR4⁺DR7⁻	—	—	—	1403	> 2000	68	—	—	—

IFNγ secretion (pg/mL) in 20 h coculture supernatants of target cells expressing HLA-A*0201, HLA-DRβ1*0401, and/or HLA-DRβ1*0701 with T lymphocytes.

^†

EBV-B cells were preincubated with 50 μg/mL MHC class II-restricted peptides, and T2 cells were preincubated with 1 μg/mL MHC class I-restricted peptides. Control peptides were the same as those in Tables 2 and 3, and relevant peptides were those indicated in Table 1 for each specific T-cell line.

^‡

DCs were matured with the indicated cytokines as described in the Materials and Methods section.

^§

Underlined values indicate that IFNγ secretion in response to DCs genetically modified with gp100 vectors, peptide-loaded T2 cells or EBV-B cells, or melanoma cell lines was ≥ 50 pg/mL and at least twice background with relevant control target cells.