title: Low prevalence of selective human leukocyte antigen (HLA)-A and HLA-B epitope losses in early-passage tumor cell lines.
abstract: The down-regulation of human leukocyte antigen (HLA) class I molecules, especially the selective down-regulation of certain allelic products, is believed to represent a major mechanism of tumor escape from immune surveillance. In the present report, an original approach is described to precisely evaluate and classify HLA class I epitope losses in 30 cancer patients with malignant melanoma and lung, breast, endometrium, ovary, and colon carcinoma tumors. Early-passage tumor cell lines were established in culture from the corresponding metastatic tumor lesions obtained in each patient. Both the cell lines and the tumor lesions were compared, in their HLA-A and -B expression, to the peripheral blood mononuclear cells (PBMCs) obtained from the same patient (autologous PBMCs). On the basis of HLA-genotyping data, the appropriate monoclonal antibodies identifying mono- and poly-morphic HLA-A and HLA-B epitopes were selected from a panel of 34 antibodies for a total of 24 testable alleles. The selected antibodies were used not only in immunohistochemical assays on cryostatic tumor sections and cytospins of PBMCs but also in quantitative, sensitive flow cytometry assays on early-passage tumor cells and PBMC suspensions. With this latter method, a low overall HLA expression was detected in 26 tumor cell explants and a complete, generalized HLA-A, HLA-B, HLA-C loss in the remaining 4 cases. However, no complete, selective loss of any of the 45 tested HLA-A and HLA-B allomorphs was observed. Sequences from all of the HLA class I alleles could be detected at the genomic DNA level in tumor cells and tissues. At variance from the literature and the results of immunohistochemical experiments performed in parallel on the corresponding tumor lesions, the relative proportions of the various HLA epitopes were relatively preserved in each early-passage cell line/PBMC pair, and selective increases, rather than decreases, in the expression of polymorphic HLA epitopes had the highest prevalence and greatest magnitude. Our data suggest an alternative tumor stealth strategy in which up- and down-regulation are equally important. This alternative model of tumor-host interaction better fits the available models of tumor cell recognition by CTLs and natural killer cells bearing activatory and inhibitory receptors for HLA-A, HLA-B, HLA-C molecules.
authors.affiliation: Laboratory of Immunology Regina Elena Institute, Centro della Ricerca Sperimentale, Rome, Italy. giacomini@crs.ifo.it
authors.forename: P
authors.initials: P
authors.lastname: Giacomini
authors.forename: E
authors.initials: E
authors.lastname: Giorda
authors.forename: R
authors.initials: R
authors.lastname: Fraioli
authors.forename: M R
authors.initials: MR
authors.lastname: Nicotra
authors.forename: N
authors.initials: N
authors.lastname: Vitale
authors.forename: A
authors.initials: A
authors.lastname: Setini
authors.forename: L
authors.initials: L
authors.lastname: Delfino
authors.forename: A
authors.initials: A
authors.lastname: Morabito
authors.forename: M
authors.initials: M
authors.lastname: Benevolo
authors.forename: I
authors.initials: I
authors.lastname: Venturo
authors.forename: M
authors.initials: M
authors.lastname: Mottolese
authors.forename: G B
authors.initials: GB
authors.lastname: Ferrara
authors.forename: P G
authors.initials: PG
authors.lastname: Natali
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D015536:Down-Regulation; D000939:Epitopes / Q000276:immunology / Q000378:metabolism*; D005434:Flow Cytometry; D005838:Genotype; D015234:HLA-A Antigens / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D015235:HLA-B Antigens / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D007157:Immunologic Surveillance; D009369:Neoplasms / Q000276:immunology*; D011110:Polymorphism, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D015234:HLA-A Antigens; D015235:HLA-B Antigens