Adoptive T Cell Therapy for Melanoma

Mice were injected subcutaneously with 1–5 × 10⁵ B16 melanoma cells and treated with intravenous adoptive transfer of freshly isolated 10^6–7 fresh splenocytes (∼2 × 10⁶ CD8⁺ Vβ13⁺ T cells) or in vitro–activated pmel-1 splenocytes (10^6–7 CD8⁺ Vβ13⁺ T cells). Mice (n = 5 for all groups) were vaccinated by intravenous injection of 2 × 10⁷ plaque-forming units of rVV or rFPV encoding mgp100, hgp100, or β-galactosidase (31 (link)) or by subcutaneous injection with 100 μl water/IFA emulsion containing 100 μg of mgp100_25–33, hgp100_25–33, or β-gal_96–103 peptide followed by two daily intraperitoneal injections of 100 μg anti-CD40 mAb purified from FGK45 hybridoma culture supernatant. rhIL-2 (a gift from Chiron Corp.) was administered intraperitoneally directly after vaccination (100,000 Cetus Units or 600,000 IU rhIL-2 in PBS, twice daily for 3–5 d). Tumors were measured with calipers and the products of perpendicular diameters were recorded. Mice were killed once tumors reached 400 mm². All experiments were performed in a blinded, randomized fashion (measuring investigator had no knowledge of the experimental group) and performed independently at least twice with similar results.