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Particulate Naturally Processed Peptides Prime a Cytotoxic Response against Human Melanoma in Vitro¹

Laboratorio di Immunoterapia Adottiva [M. P. P., M. A. I., A. A. M., G. Co., S. H., M. B., C. R.], Laboratorio di Tecniche Separative [C. A.], and Unità di Immunochimica, Departimento Biotecnologie [P. D., G. Ca.], Istituto Scientifico H. San Raffaele, via Olgettina 60, 20132 Milan, and Cattedra di Medicina Interna II, Università degli Studi di Milano, 20100 Milan [C. R.], Italy

For an efficient antitumor cytotoxic response, tumor antigenic peptides need to be presented by professional antigen-presenting cells in association with MHC class I molecules. We established in vitro short-term human CTL lines from healthy and melanoma-bearing subjects, using as antigen-presenting cells autologous adherent cells after phagocytosis of latex beads coated with melanoma peptides. Melanoma peptides were obtained by acid extraction of melanoma cells that matched with donor peripheral blood mononuclear cells, at least for one HLA-A allele. The cytotoxic activity of the lines was specific for the melanoma from which peptides were obtained and for melanoma sharing HLA alleles. These results demonstrate that a complex mixture of naturally processed melanoma peptides conjugated to a phagocytic substrate that targets them into the MHC class I pathway of adherent cells can prime a CTL response in healthy subjects in vitro, and that peptides from allogeneic tumors may be used to propagate CTL in melanoma patients. Our data support the feasibility of active and passive vaccination procedures with nonliving vaccines in cancer patients.

¹ This work was supported by the Fondazione Centro San Raffaele del Monte Tabor, by Progetto Finalizzato Applicazioni Cliniche Ricerca Oncologica Grant 9202374 PF39 from the Consiglio Nazionale delle Ricerche (to C. R.), and by Grant 324/95 from the Associazione Italiana Ricerca sul Cancro (to C. R.). P. D. and G. C. are supported by the Associazione Italiana Ricerca sul Cancro.

² To whom requests for reprints should be addressed.

Received 11/27/95. Accepted 1/29/96.

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