Treatment of hepatocellular carcinoma in a mouse xenograft model with an immunotoxin which is engineered to eliminate vascular leak syndrome.
Vascular leak syndrome (VLS) is the major dose-limiting toxicity of immunotoxin and interleukin-2 therapy. It has been evidenced that VLS-inducing molecules share a three-amino acid consensus motif, (x)D(y), which may be responsible for initiating VLS. Here we have constructed a recombinant immunotoxin (SMFv-PE38KDEL) by genetically fusing PE38KDEL to a single-chain antibody derived from SM5-1 monoclonal antibody, which has a high specificity for melanoma, hepatocellular carcinoma and breast cancer. In order to eliminate VLS induced by this PE38KDEL-based immunotoxin, a panel of mutants were generated by changing amino acid residues adjacent to its three (x)D(y) motifs in the three-dimensional structure. One of the SMFv-PE38KDEL mutants, denoted as mut1, displayed a similar protein synthesis inhibitory in a reticulocyte lysate translation assay compared to the wild-type SMFv-PE38KDEL (wt). The in vitro cytotoxicity assay indicated that mut1 specifically killed SM5-1 binding protein-positive tumor cells, although its cytotoxicity was slightly less than wt. In contrast, mut1 was shown to be much weaker in inducing VLS in mice than wt. The LD(50) values of wt and mut1 in mice were investigated with the result that the LD(50) of mut1 was about tenfold higher than that of wt. The in vivo antitumor activity of wt and mut1 were also compared in tumor-bearing nude mice. Both wt and mut1 were effective in inhibiting the tumor growth but mut1 showed improved therapeutic efficacy. These studies suggest mut1 may be a novel PE-based immunotoxin with much less toxicity for clinical use.
Wang
Hao
H
International Joint Cancer Institute, Second Military Medical University, Shanghai, PR China.
Song
Shuichuan
S
Kou
Geng
G
Li
Bohua
B
Zhang
Dapeng
D
Hou
Sheng
S
Qian
Weizhu
W
Dai
Jianxin
J
Tian
Liang
L
Zhao
Jian
J
Guo
Yajun
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology; D019559:Capillary Leak Syndrome / Q000150:complications / Q000517:prevention & control*; D006528:Carcinoma, Hepatocellular / Q000628:therapy*; D004195:Disease Models, Animal; D005818:Genetic Engineering; D015316:Genetic Therapy; D000922:Immunotoxins / Q000235:genetics* / Q000494:pharmacology*; D008114:Liver Neoplasms, Experimental / Q000628:therapy*; D051379:Mice; D008819:Mice, Nude; D008958:Models, Molecular; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology*; D014183:Transplantation, Heterologous
D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D000922:Immunotoxins; D011994:Recombinant Proteins