A pilot Phase I study combining peptide-based vaccination and NGR-hTNF vessel targeting therapy in metastatic melanoma

Figures & data

Table 1. T cell-mediated immune response of patients enrolled in the trial

Patient # and peptide vaccine	Recognition of APCs loaded with indicated melanoma peptides	Recognition of melanoma cell lines
# 02/NA17	No	Yes; No HLA restriction
#03/NA17	N0	Yes; No HLA restriction
#04/NA17	Yes (MAGE-A2, gp100 not NA17)	Yes; No HLA restriction
#05/NA17	Yes (NA17, MAGE-A2, MAGE-A3)	Yes; No HLA restriction
#06/MAGE-A3	Yes (MAGE-A2, MAGE-A3, NA-17, NY-ESO-1)	Yes; HLA-restriction
#07/NA17, MAGE-A3	Yes, (NA17, MAGE-A2, MAGE-A3)	Yes; HLA-restriction
#08/NA17	Yes (NA17, MAGE-A2, MAGE-A3, NY-ESO1, Tyr)	Yes; HLA restriction

Figure 1. T-cell responses to MAAs in PBMCs of melanoma patients undergoing NGR/VAX treatment. Freshly isolated peripheral blood mononuclear cells (PBMCs) from melanoma patients (#02, 05, 07 and 08) were used to assess their reactivity against melanoma associated antigens (MAA)-derived peptides (MAGE-A2, MAGE-A3, NA-17A, NY-ESO-1, MART-1, gp100, TYR) on HLA-A*01⁺ 1061 EBV-B (Panel A and D) or HLA-A*0201⁺ T2 cells (Panels A, B and C) and autologous, when available, or allogeneic HLA-matched tumor cell reactivity (Panel C). Interferon γ (IFNγ)-based ELISPOT assay was used for this analysis. Data are expressed as N. of spots/3 × 10⁴cells and are subtracted of the background of IFNγ release from T cells incubated with EBV-B or T2 control cells alone. Results represent averages of triplicates with SD ≤ 10%; statistical analysis of differences between means of IFNγ released by T cells was performed by 2-tailed Student's t-test; significance defined as p < 0.05.

Table 2. Clinical outcome of melanoma patients treated with NGR/Vax/01

Patient #	Stage	Date of treatment onset	Additional therapy	OS^§§	TTP
01	IV M1c	25/11/2010	Zelboraf*	11	NA
02	III M1a	01/03/2011	Surgery** Chemotherapy** IL-2** Vaccination** (2006)	29+SD	26
03	IV M1c	01/07/2011	Surgery** Chemotherapy** Ipilimumab*,Local RT*	4	1.3
04	IV M1b	22/07/2011	Chemotherapy** IFNα**	7	1
05	IV M1c	28/7/2011	Ipilimumab** Chemotherapy**	4	4
06	IV M1a	10/10/2011	Surgery**	25+NED	11
07	IV M1a	14/11/2011	Surgery*** RT*	25+NED	3.3
08	IV NED	19/12/2011	Surgery*** RT*	23+NED	1

*After drop out; **Before protocol treatment; ***During protocol treatment; §: Overall Survival (OS) from Protocol Start (months). RT: Radiotherapy; SD: Stable disease; NED: No evidence of disease.

Figure 2. Detection of TNF inhibitory soluble factors and VEGF in the serum of melanoma patients. Serum from melanoma patients was collected at different time points: baseline, 2 hours, 1 month and 2 months post-treatments. The presence in the serum of chromogranin A (CgA), soluble tumor necrosis factor (sTNFR) was assessed by ELISA. Statistical analysis of differences between means of soluble factors was performed by 2-tailed Student's t-test with significance defined as p < 0.05.

Figure 3. Detection of pro-inflammatory factors (MIP1β and MCP1) in the serum of melanoma patients. Serum from melanoma patients was collected at different time points: baseline, 2 hours, 1 month and 2 months post-treatments. The presence in the serum of macrophage chemoattractant protein 1 (MCP-1) and macrophage inflammatory protein 1β (MIP-1β) was assessed by ELISA. Data represent pg/mL of each soluble factor. Statistical analysis of differences between means of soluble factors was performed by 2-tailed Student's t-test with significance defined as p < 0.05.