title: Direct transfer of a foreign MHC gene into human melanoma alters T cell receptor V beta usage by tumor-infiltrating lymphocytes.
abstract: The direct introduction of foreign genes into tumors shows promise as a therapeutic modality to enhance tumor immunogenicity. Hence, melanoma nodules were directly injected with a vector encoding an allogeneic MHC class I molecule, HLA-B7. Tumor-infiltrating lymphocytes (TIL) were isolated from cutaneous melanoma biopsies before and after HLA-B7 gene transfer. TIL were expanded in interleukin-2 (IL-2) by standard techniques for approximately 4 weeks, then analyzed for T cell receptor V beta usage by quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Prior to gene transfer. TIL V beta usage was found to be highly restricted, the only one to four V beta families being expressed and one or two of these families representing more than 90% of the repertoire. As anticipated, TIL V beta usage varied among patients expressing different HLA types. However, V beta 13 was over-represented in that six of eight patients utilized V beta 13 as a dominant family, regardless of HLA type. Following HLA-B7 gene transfer, TIL V beta usage was markedly altered: (1) V beta families that dominated following gene transfer differed from the V beta families utilized by TIL prior to treatment, and (2) introduction of the HLA-B7 gene resulted in a more diverse repertoire with an increase in the number of V beta families represented. In two patients, TIL were evaluated before treatment and from multiple, distinct melanoma nodules following gene transfer. In these two patients, a comparison was made between TIL V beta profiles obtained after treatment from nodules that had been injected with the HLA-B7 gene or left untreated. Interestingly, the V beta repertoires of TIL from uninjected nodules following gene transfer were similar to that of TIL from injected nodules, rather than pretreatment TIL. These data demonstrate a direct immunological effect of foreign MHC gene transfer into human melanoma, and suggest that local expression of an allogeneic MHC molecule generates systemic alterations in the antitumor immune response.
authors.affiliation: Section of Thoracic Surgery, University of Michigan School of Medicine, Ann Arbor 48109, USA.
authors.forename: L A
authors.initials: LA
authors.lastname: DeBruyne
authors.forename: A E
authors.initials: AE
authors.lastname: Chang
authors.forename: M J
authors.initials: MJ
authors.lastname: Cameron
authors.forename: Z
authors.initials: Z
authors.lastname: Yang
authors.forename: D
authors.initials: D
authors.lastname: Gordon
authors.forename: E G
authors.initials: EG
authors.lastname: Nabel
authors.forename: G J
authors.initials: GJ
authors.lastname: Nabel
authors.forename: D K
authors.initials: DK
authors.lastname: Bishop
mesh_terms: D015704:CD4 Antigens / Q000502:physiology; D002469:Cell Separation; D018014:Gene Transfer Techniques; D018740:Genetic Heterogeneity; D015235:HLA-B Antigens / Q000235:genetics*; D006801:Humans; D016246:Lymphocytes, Tumor-Infiltrating / Q000737:chemistry* / Q000166:cytology / Q000276:immunology; D008545:Melanoma / Q000235:genetics* / Q000473:pathology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000378:metabolism*
chemical_list: D015704:CD4 Antigens; D015235:HLA-B Antigens; D016693:Receptors, Antigen, T-Cell, alpha-beta