HLA TYPING FOR IMMUNOTHERAPY BY MICROCHIPS.
G. B. Ferrara*, L. Delfino*, G. Salani°, G. L. De Bellis° *IST c/o Advanced Biotechnology Center, Genova, Italy, °I.T.B.A., CNR, Segrate, MI, Italy.


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The HLA typing of patients is extremely important for a correct vaccinotherapy (i.e. melanoma) with peptides derived from tumoral antigens of the recipient and presented to T cells by different types or subtypes of the same HLA alleles. We have developed a microchip that can type the patients for the HLA antigens bound by peptides for an efficient therapy. The microchip can define the following antigens, HLA-A1, all A2 subtypes detected so far, A6802, A3, A3.1, A31, B44, Cw6, Cw16 which cover most of the specificities known to be used by tumor derived peptides. Several technical approaches were devised: oligonucleotides were either spotted on silane treated glass or on streptavidin coated inert surfaces by single or multiple silica needle robotized via an Hamilton 2200. Oligo microarrays were then used for hybridization or for minisequencing experiments. Signal detection was performed either by evanescent wave fluorescence or time resolved fluorescence detected using an intensified CCD or a cooled CCD. Chemiluminiscence was tested in parallel. Results will be shown demonstrating the quality of these results.