Inoculation site from a cutaneous melanoma patient treated with an allogeneic therapeutic vaccine: a case report.

Mariana Aris,María Marcela Barrio,José Mordoh,Alicia Inés Bravo

doi:10.3389/fimmu.2015.00144

Abstract

We have developed a therapeutic vaccine consisting of a mixture of lethally-irradiated allogeneic cutaneous melanoma cell lines with BCG and GM-CSF as adjuvants. The CSF-470 vaccine is currently being assayed in a Phase II–III trial against medium-dose IFN-α2b. All vaccinated patients immunized intradermally developed large edematous erythema reactions, which then transformed into subcutaneous nodules active for several months. However, vaccine injection sites were not routinely biopsied. We describe the case of a female patient, previously classified as stage III, but who, due to the simultaneous discovery of bone metastases only received one vaccination was withdrawn from the study, and continued her treatment elsewhere. This patient developed a post-vaccination nodule which was surgically removed 7 weeks later, and allowed to analyze the reactivity and immune profiling of the inoculation site. An inflammatory reaction with zones of fibrosis, high irrigation, and brisk lymphoid infiltration, primarily composed of CD8+ and CD20+ lymphocytes, was observed. There were no remaining BCG bacilli, and scarce CD4+ and Foxp3+ T cells were determined. MART-1 Ag was found throughout the vaccination site. CD11c+ Ag presenting cells were either dispersed or forming dense nests. Some CD11c+ cells proliferated; most of them contained intracellular MART-1 Ag, and some interacted with CD8+ lymphocytes. These observations suggest a potent, long-lasting local inflammatory response with recruitment of Ag-presenting cells that incorporate melanoma Ags, probably leading to Ag presentation to naïve T cells.

Highlights

A 45-year-old Caucasian woman underwent resection on May 2012 of a spontaneously bleeding congenital nevus in her back
We have developed a therapeutic vaccine consisting of a mixture of lethally-irradiated allogeneic cutaneous melanoma cell lines with bacillus calmette guerin (BCG) and GM-CSF as adjuvants
The classic paradigm proposes that Ag presentation occurs exclusively in secondary lymph nodes (LN), evidence from afferent lymph vessels in normal human skin revealed an increased number of mainly memory/effector CD4+CD45Ro+ T cells and IL-12+ dendritic cells (DC) in contact with IFN-α-producing T cells, supporting that T cells may be stimulated by Ag presenting cells (APC) in peripheral tissues [10]

Summary

INTRODUCTION

A 45-year-old Caucasian woman (patient #1) underwent resection on May 2012 of a spontaneously bleeding congenital nevus in her back. Histological analysis revealed an ulcerated nodular cutaneous melanoma (CM) with a Breslow thickness of 7.8 mm, epithelioid and spindle cell morphology, non-brisk immune infiltration, and satellitosis. Dense nested structures comprised of macrophages, histiocytes and polynuclear cells, typically found in inflammatory processes including responses to BCG, were observed in the highly infiltrated zone (Figure 1C). Immune profiling analysis revealed brisk CD8+ and CD20+ lymphocyte infiltration, nonbrisk CD4+, and scarce Foxp3+ T cells infiltration (Figures 2A– D,G). Brisk infiltration of CD11c+ Ag-presenting cells (APCs) was observed; these cells were either dispersed in the dermis or assembled in multiple dense nested structures (Figures 2F,G). Revealed that some CD11c+ APCs, both dispersed and nested, proliferated (Figures 3A,B,G), most had phagocytosed MART-1 Ag (Figures 3C,D,G), and some were surrounded by CD8+ lymphocytes (Figures 3E,F), suggesting local Ag presentation. The near absence of FoxP3+ lymphocytes suggests that an immunogenic environment was created

BACKGROUND

DISCUSSION