Hajime Kawaharada

USEFULNESS OF GLYCOSYLATED RECOMBINANT HUMAN LYMPHOTOXIN FOR GROWTH INHIBITION OF HUMAN AND MURINE SOLID TUMORS AND EXPERIMENTAL METASTASIS IN MICE

Summary We have examined the antitumor and antimetastatic effects of native- type, glycosylated recombinant lymphotoxin (LT) on human and murine tumors transplanted in mice. The results reported here are as follows: (a) The in vivo antitumor spectrum of LT is not coincident with the in vitro study, and it has a wide antitumor spectrum and substantially inhibits the growth of human solid tumors, (b) When both syngeneic and nude mice are transplanted with Meth A tumor, the significant growth-inhibitory effect of LT is obtained in syngeneic mice, but the effect is quite small in nude mice regardless of the routes; LT attains the same degree of effectiveness as that in syngeneic mice, but at an 8 to 16 times higher dose. Furthermore, the pretreatment with antiasialo- GMl antibody inhibits the antitumor effects of LT in syngeneic mice, (c) In the pulmonary metastasis model induced by i.v. injection of Meth A cells, a high preventive effect of LT is obtained by systemic administration in syngeneic mice, but not in nude mice. In addition, the pretreatment with antiasialo- GMl antibody completely prevents the antimetastatic effect of LT, but also blocks that effect of control mice without LT treatment. In conclusion, LT appears to be a potent cytokine against tumor growth and metastasis in vivo. The differences between nude and syngeneic mice suggest the involvement of host immunity in the expression of LT function.

Comparison of Human Lymphotoxin Gene Expression in CHO Cells Directed by Genomic DNA or cDNA Sequences

Four recombinant plasmids coding for human lymphotoxin (LT) were constructed with genomic DNA (gDNA) or cDNA sequences. The simian virus 40 (SV40) early region, which contains the early promoter, an intron of the small-t-antigen-encoding gene, and polyadenylation signal sequences, was used for transcriptional and post-transcriptional regulatory elements in the construction of these plasmids. Two of them contained gDNA and the other two contained cDNA. One of the gDNA plasmids and one of the cDNA plasmids carry the SV40 intron between the structural gene and polyadenylation signal. Transient and stable gene expression levels of these plasmids in Chinese hamster ovary (CHO) cells were measured by assaying the secreted LT. The plasmid carrying gDNA without the SV40 intron was expressed more efficiently than the other three plasmids in both transient and stable gene expression assays.