"Biological effects of soluble recombinant CD40 ligand and interferon-γ on human renal cancer cell lines in vitro and in vivo."

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1. Surface expression of CD40 on human renal cancer cell line.Immunofluorescence studies were performed by flow cytometric analysis. CD40 was expressed on various human renal cancer cell lines, 769-P, 786-O, A-498 and A-704.2. Induction of CD40 expression by interferon-γ in vitro.Incubation with interferon-g significantly induced surface expression on human renal cancer cell lines.3. Antitumor effects of soluble recombinant CD40 lingand (srCD40L) and interferon-γ on human renal cancer cell lines in vitro.Incubation with srCD40L significantly inhibited the proliferation of 769-P and 786-O as determined by the proliferation assay with optimal inhibition of thymidine incorporation. Apoptotic studies by flow cytometry revealed an induction of apoptosis in these two cell lines. Additional inhibitory effect by co-incubation with interfron-γ significantly enhanced this inhibitory effects.4. Antitumor effects of srCD40L treatment in SCID mice bearing human human renal cancer cell lines.Effects of srCD40L treatment on survival in human renal cancer cell-bearing SCID mice were determined. All mice received 20 μL of anti-asialo GMI (Wako Chemicals, Osaka) by intravenous injection (IV) 1 day before tumor transfer to remove host natural killer cells. 769-P cells (5 x l0ィイD16ィエD1) were then administered by IV. SCID recipients then received either l0 μg of srCD40L or controll every other day for 20 days for total of 10 injections starting at day 3. Tumor bearing mice were then monitored for tumor development and progression. Treatment with srCD40L significantly improved the survival of tumor-bearing mice (p<0.05).We are preparing a manuscript from these data for Japanese Journal for Cancer Research.