Lymphokine-mediated immunotherapy studies in mouse tumor systems

Abstract

Studies have been performed using two subcutaneously implanted mouse tumor models to investigate the immunotherapeutic potential of lymphokine-containing culture supernatants from long-term human lymphoblast cell cultures. Human lymphoblastoid cell line, RPMI 1788, was used as a cell culture source of lymphokines. Supernatants were removed from cultures at the stationary phase of growth and concentrated on Amicon filters retaining molecules above 10,000 Daltons. This concentrate was applied to a Sephadex G-25 column, equilibrated with ammonium bicarbonate buffer, for removal of salts and dye from the culture medium. The effluent was lyophilized and reconstituted for use in further purification by affinity chromatography and SDS-PAGE gels. Such preparations were used to inject DBA/2 mice bearing subcutaneous L-1210 tumors. In addition, the B-16 melanoma was used as a model of a solid tumor in C57Bl/l mice. Animals were treated intralesionally and intraperitoneally with lymphokines containing preparations and control solutions. Tumors growing subcutaneously were susceptible to lymphokine-induced inflammation-mediated regression without additional therapy. In the study of L-1210 subcutaneous tumors, reduction in tumor size was followed by complete regression, prolonged survival, immunity to additional inoculation, and cures in 20--40% of the treated mice. Tumor regression and prolongation of survival were also noted in mice bearing B-16 melanomas. These studies support the use of mouse tumors as bioassays for antitumor inflammatory activity of human lymphokine preparations and help to quantitate their potential use in human tumor immunotherapy.

Tumour growth inhibition in mice by glycosylated recombinant human lymphotoxin: analysis of tumour-regional mononuclear cells involved with its action

“…It has also been reported that purified recombinant human TNF shows efficacy against tumours in vivo (Sohmura et al, 1986;Palladino, 1987;Manda et al, 1987;Asher et al, 1987;Watanabe et al, 1988;Inagawa et al, 1988). However, there are few published data for in vivo studies of glycosylated human LT(gLT), (Khan et al, 1982;Papermaster et al, 1980;Ransom et al, 1982), because the production of recombinant LT with sugar moieties was not achieved until quite recently. Human lymphotoxin genomic DNA has recently been cloned, and an efficient system for producing human gLT in Chinese hamster ovary cells has now been established in our laboratory (Nakagawa et al, 1991).…”

Effect of Delayed-Type Hypersensitivity Reaction and Transferred Lymphokine on the Resistance of Mice to Salmonella typhimurium Infection

“…Several studies which used lymphokine(s) in the treatment of murine tumors have been reported (6,13,15). However, little attention has been paid to the in vivo applicability of lymphokine(s) in the control of infection with intracellular parasites.…”