Early effects of CSF-470 melanoma vaccine plus adjuvants in immune cell stimulation

PAMPENA, MARÍA BETINA; JULIÁ, ESTEFANÍA PAULA; ROCCA, YAMILA SOL; BLANCO, PAULA ALEJANDRA; BARRIO, MARÍA MARCELA; MORDOH, JOSÉ; LEVY, ESTRELLA MARIEL

Congreso; AACR Annual Meeting 2017; 2017

Background: In a Phase II/III clinical trial (CASVAC-0401, NCT01729663), cutaneous melanoma (CM) patients (pts) stages II/III were randomized to receive CSF-470 vaccine or IFN-α (2:1). Vaccine CSF-470 is composed of four lethally-irradiated allogeneic CM cell lines; BCG (Bacillus Calmette-Guérin) and rhGM-CSF were used as adjuvants. At the end of the Phase II part of the study (30 pts), a higher distant-metastases-free-survival was observed in vaccinated pts. All vaccinated pts developed inflammation at the vaccination site. Therefore, in an attempt to understand these first steps of immune reaction, leukocytes migration, cytokines synthesis and monocyte phagocytosis promoted by the vaccine components were studied in vitro.Methods: Cell migration of healthy donors? leukocytes or mononuclear cells (MN) towards vaccine components was assayed in 0,5 μm transwells. Cytokines were measured by ELISA in supernatants of CSF-470 vaccine and MN (1:1, 0,5x106 each) cocultures. For phagocytosis assays, CM cell lines were labeled with PKH67 lipophilic dye, irradiated, and cocultured with purified monocytes. After 24 hs, cultures were stained with CD14 and double staining (CD14+PKH67+) was analyzed by flow cytometry.Results: In migration assays, CSF-470 vaccine induced higher attraction of leukocytes as compared to culture medium (p=0.003, unpaired-T-test). GM-CSF exerted little attraction, although combined with BCG it increased the migration triggered by CSF-470 (vaccine vs vaccine plus adjuvants, p=0.034, unpaired-T-test). When using MN, CSF-470 alone or plus adjuvants, highly attracted monocytes, which represented 10%-65% of the migrating CD45+ population (before migration 6-15%) (p