Liver and tumour tissue concentrations of TNF-alpha in cancer patients treated with TNF-alpha and melphalan by isolated liver perfusion

Pjk Kuppen,Cjh Van De Velde,Ihm Borel Rinkes,Amm Eggermont,Rame Tollenaar,Le Jonges,Al Vahrmeijer

doi:10.1038/bjc.1997.255

Abstract

In this study we determined the level of tumour necrosis factor alpha (TNF-alpha) in liver and tumour tissue samples obtained from patients with colorectal metastases confined to the liver, who were treated with isolated liver perfusion with TNF-alpha and melphalan. We adapted a standard enzyme-linked immunosorbent assay kit for the quantification of TNF-alpha in serum to measure the amount of this cytokine in solid tissue. For this purpose, we developed a buffer that lysed the tissues without affecting the TNF-alpha present. The minimum detection level was about 2 pg of TNF-alpha per mg tissue. Using this technique, we found a significant increase in the TNF-alpha level after perfusion in the liver tissue of all evaluable patients, which may explain the transient liver toxicity we observed in all patients. In tumour tissue, a significant TNF-alpha increase was observed in one out of five patients. The level of TNF-alpha in all liver tissue samples and some of the tumours after treatment by isolated liver perfusion was much higher than the peak serum concentrations obtained after systemic administration of the maximum tolerated dose of TNF-alpha. Furthermore, we demonstrated that the level of TNF-alpha in the liver tissue samples was about seven to eight times higher than in tumour tissue. We concluded that regional liver treatment resulted in a relatively high local level of TNF-alpha, but also that this cytokine did not preferentially accumulate in tumour tissue.

Highlights

A standard enzyme-linked immunosorbent assay (ELISA) kit for the detection of tumour necrosis factor alpha (TNF-a) in serum was used for the determination of TNF-ax in liver and tumour biopsies
Because these biopsies had to be solubilized before they could be used in the ELISA, the influence of the lysis buffer used for this purpose on the detection level of TNF-a was tested
This was done by the addition of known concentrations of TNF-a to the lysis buffer and subsequent measurement of the TNF-a level in this solution by the ELISA to determine the recovery

Summary

Methods

Biopsies of liver and tumour tissue of six patients were taken just before and directly after an isolated liver perfusion for 1 h with either 0.4 (n = 5) or 0.8 (n = 1) mg of human recombinant TNF-a (Boehringer-Ingelheim, Ingelheim am Rhein, Germany) together with melphalan (L-PAM, 1 mg kg-' body weight; Wellcome, Beckenham, Kent, UK). None of the patients had other known liver aberrations in their past history. During the isolated liver perfusion, the liver was drained for 1 h with TNF-a and melphalan. The inflow from the isolated circuit ran via the common hepatic artery and the portal vein. The hepatic venous outflow returned the perfusate to the oxygenator. The intestinal, renal and lower extremity blood was shunted around the liver and brought back to the heart via the axillary vein (Vahrmeijer et al, 1995)

Results

Discussion

Conclusion