Anti asialo gm1 antiserum

Athymic female mice (6–8 weeks old) (Harlan Sprague-Dawley) were housed in the FHCRC animal facility according to the Institutional Animal Care and Use Committee. All mice were placed on a biotin-deficient diet (Purina Mills) 7 days prior to PRIT studies. Ramos, Granta-519, or Granta-519^luc cells (10⁷) were injected subcutaneously in the right flank 7–14 days prior to experiments to produce lymphoma xenografts measuring 6 to 8mm in diameter. Anti-asialoGM1 antiserum (30 μl, WAKO) was injected i.p. 8 days and 3 days prior to FP injection, and weekly for 100 days to prevent spontaneous tumor regressions.

The subline F10.P1 of the B16 melanoma (C57BL/6 origin) was established in our laboratory from a B16-F10 lung metastasis. The chicken OVAlbumin-transduced M05 variant of the B16 melanoma cell line (expressing the SIINFEKL peptide in H-2Kb) was a kind gift from Dr. Louis Falo, University of Pittsburgh [40 (link)]. Lewis lung carcinoma (3LL) and Panc02 adenocarcinoma cells were purchased from The American Type Culture Collection (ATCC). The MC38 colon carcinoma was a gift from Dr. M. Shurin, University of Pittsburgh. MC38 and Panc02 tumor cells were transfected to produce OVA-expressing variants, MC38^OVA and Panc02 ^OVA, respectively. All cell-lines were maintained in RPMI-1640 medium (Life Technologies, Gaithersburg, MD, USA) supplemented with 10% heat inactivated fetal calf serum, 2 mM glutamine, 20 mM Hepes buffer, 0.8 g/l streptomycin and 1.6×10⁵ U/l penicillin (from hereon referred to as complete medium, CM). Adherent cells were detached by exposure to 0.02% EDTA for 2–3 min and washed three times in RPMI-1640. Cell viability, judged by trypan blue dye exclusion test, was always >95%. Murine pulmonary metastases were established by tail vein injection of 0.2–0.4×10⁶ cells in 0.3 ml of RPMI-1640 into C57BL/6 mice, pretreated on day −1 with 40 μl anti-asialoGM1 antiserum (Wako Pure Chemicals, Wako, TX, USA).