A B7‐1‐transfected human melanoma line stimulates proliferation and cytotoxicity of autologous and allogeneic lymphocytes

Abstract

B7 co-stimulation is necessary to activate resting T cells upon antigen recognition by the T cell receptor. To see whether expression of B7 may render human melanoma cells able to stimulate T cells, a cloned melanoma line (Me1B6), which did not express B7-1, was transfected with the human B7-1 gene. In proliferation assays, B7-1 transfected cells (Me1B6/B7) showed greater stimulatory activity of allogeneic and autologous peripheral blood lymphocytes (PBL) compared to parental, non-transfected tumor cells. This effect was also seen when allogeneic CD8+ and CD4+ subpopulations were used as effectors. In these studies, activation of lymphocytes was B7-1-dependent and HLA classes I and II mediated. The higher proliferation correlated with an increased lytic activity by PBL stimulated with B7-1+ tumor cells against the untransfected Me1B6. Furthermore, PBL from a metastatic melanoma patient stimulated by Me1B6/B7 developed an higher lytic activity not only against Me1B6 but also against their autologous, B7-1- tumor. Finally, after Me1B6/B7 stimulation, PBL released interleukin (IL)-2 and interferon-gamma, but not IL-4, suggesting a Th1-mediated response. These data support the use of B7-1 transfected melanoma cells in the therapeutic vaccination of melanoma patients.

Type I consensus interferon (CIFN) gene transfer into human melanoma cells up-regulates p53 and enhances cisplatin-induced apoptosis: implications for new therapeutic strategies with IFN-alpha

“…Isolation and in vitro characterization of IFN-producing human melanoma cell lines The human melanoma cell lines 8863 (metastatic melanoma) 20 and 1B6 (primary melanoma) 21 were infected with the recombinant retroviruses LXSN and LCIFNSN, derived from the retroviral vectors schematically depicted in Figure 1. Following selection into G418-containing medium, cell lines producing approximately 1000 IU/ml of IFN were isolated from both 8863 and 1B6 cells (Table 1).…”

“…20 The HLA haplotype of the two patients, previously identified, was: patient 10538: HLA-A2, -A28, -B27, -B45, -C-, -C-, -DR7; patient 8863: HLA-A2, -A1, -B8, -B35, -Cw4, -C-, -DR3, -DR11. 20 The 1B6 and 8863 cell lines had been previously shown not to express the B7.1 co-stimulatory molecule. 20 Both cell lines were cultivated in RPMI-1640 supplemented with 50 u/ml penicillin, 50 g/ml streptomycin (BioWhittaker, Verviers, Belgium), and 10% FCS (Sebam, Berlin, Germany).…”

“…20,21 The 1B6 clone was isolated from the Me10538 cell line, established from a primary skin melanoma lesion, 21 whereas the 8863 cell line was derived from a metastatic melanoma. 20 The HLA haplotype of the two patients, previously identified, was: patient 10538: HLA-A2, -A28, -B27, -B45, -C-, -C-, -DR7; patient 8863: HLA-A2, -A1, -B8, -B35, -Cw4, -C-, -DR3, -DR11. 20 The 1B6 and 8863 cell lines had been previously shown not to express the B7.1 co-stimulatory molecule.…”

“…20 The 1B6 and 8863 cell lines had been previously shown not to express the B7.1 co-stimulatory molecule. 20 Both cell lines were cultivated in RPMI-1640 supplemented with 50 u/ml penicillin, 50 g/ml streptomycin (BioWhittaker, Verviers, Belgium), and 10% FCS (Sebam, Berlin, Germany).…”

Adenoviral delivery of B7–1 (CD80) increases the immunogenicity of human ovarian and cervical carcinoma cells

“…These vectors may be suitable for modification of tumour cells without extended culture ex vivo as Jaffee et al 25 reported high efficiency retroviral gene transfer into a number of primary human tumour explants without drug selection. Sule-Suso et al 26 …”