Plasma Cytokine Analysis in Patients with Advanced Extremity Melanoma Undergoing Isolated Limb Infusion

Abstract

Background Preprocedure clinical and pathologic factors have failed to consistently differentiate complete response (CR) from progressive disease (PD) in patients after isolated limb infusion (ILI) with melphalan for unresectable in-transit extremity melanoma. Methods Multiplex immunobead assay technology (Milliplex MAP Human Cytokine/Chemokine Magnetic Bead Panel, Millipore Corp., Billerica, MA; and Magpix analytical test instrument, Luminex Corp., Austin, TX) was performed on pre-ILI plasma to determine concentrations of selected cytokines (MIP-1α, IL-1Rα, IP-10, IL-1β, IL-1α, MCP-1, IL-6, IL-17, EGF, IL-12p40, VEGF, GM-CSF, and MIP-1β) on a subset of patients (n = 180) who experienced CR (n = 23) or PD (n = 24) after ILI. Plasma from normal donors (n = 12) was also evaluated. Results Of 180 ILIs performed, 28 % (95 % confidence interval 22–35, n = 50) experienced a CR, 14 % (n = 25) experienced a partial response, 11 % (n = 21) had stable disease, 34 % (n = 61) had PD, and 13 % (n = 23) were not evaluable for response. Tumor characteristics and pharmacokinetics appeared similar between CR (n = 23) and PD (n = 24) patients who underwent cytokine analysis. Although there were no differences in cytokine levels between CR and PD patients, there were differences between the melanoma patients and controls. MIP-1α, IL-1Rα, IL-1β, IL-1α, IL-17, EGF, IL-12p40, VEGF, GM-CSF, and MIP-1β were significantly higher in normal controls compared to melanoma patients, while IP-10 was lower (p <0.001) in controls compared to melanoma patients. Conclusions Patients with unresectable in-transit melanoma appear to have markedly decreased levels of immune activating cytokines compared to normal healthy controls. This further supports a potential role for immune-targeted therapies and immune monitoring in patients with regionally advanced melanoma.

Serum Cytokinome Profile Evaluation: A Tool to Define New Diagnostic and Prognostic Markers of Cancer Using Multiplexed Bead-Based Immunoassays

“…Moreover, IFN-alpha2 β therapy resulted in a significant decrease of serum levels of immunosuppressive and tumor angiogenic/growth stimulatory factors (VEGF, EGF, and HGF) and increased levels of IP-10 and IFN- α [40]. Moreover, Shetty et al (2013) evidenced that the levels of MIP-1 α , IL-1R α , IL-1 β , IL-1 α , IL-17, EGF, IL-12p40, VEGF, GM-CSF, and MIP-1 β were significantly higher in normal controls compared to melanoma patients, while IP-10 level was lower [64]. …”

Antitumor Immunity Triggered by Melphalan Is Potentiated by Melanoma Cell Surface–Associated Calreticulin

“…Considering that immune responses following locoregional therapy can differ from those after systemic therapy (20), it is necessary that anticancer immunity, danger signaling, and immune-effector function-potentiating effects of locoregionally applied chemotherapeutics are also evaluated-a knowledge that is largely missing and could have translational significance (20,21).…”

Immunotherapy Following Regional Chemotherapy Treatment of Advanced Extremity Melanoma

“…19 Patients with intransit melanomas have been demonstrated to be globally immunosuppressed, suggesting a role for immunotherapy to stimulate immune response to more normal levels. 20 Within the tumor microenvironment, melphalan-based RC has been shown to upregulate the costimulatory molecules CD80 and CD86 to produce T-cell stimulation in response to tumor antigen. 21 Melphalan-based RC also has been reported to alter T-cell stimulation, NK cell activity, and sICAM-2 levels in the peripheral blood.…”