Safety evaluation of Ad5CMV-p53 in vitro and in vivo.
In preparation for a clinical trial of the recombinant p53 adenovirus Ad5CMV-p53 for the treatment of lung cancer, the potential adverse effects of Ad5CMV-p53 were assessed in vitro and in vivo. No infectious replication of Ad5CMV-p53 was detectable in HeLa cells infected with extracts from HeLa cells previously infected with Ad5CMV-p53. No Ad5CMV-p53 DNA replication was detected by 32Pi labeling in lung cancer cells infected with Ad5CMV-p53 at multiplicities of infection (moi) up to 1,000 pfu/cell (total of 5 x 10(9) pfu viruses). The infectivity and cytotoxicity of Ad5CMV-p53 were examined in vitro in normal human bronchial epithelial (NHBE) cells. At a moi of 50 pfu/cell, Ad5CMV-p53 infection and expression were detectable in 80% of the treated cells. The exogenous p53 protein was first detected by western blotting at 8 hr and peaked at 48 hr after infection. Growth of NHBE cells was not affected by Ad5CMV-p53 infection at a moi of 100 pfu/cell. The pathogenicity of Ad5CMV-p53 was assessed in BALB/c mice. The virus was given to four groups of mice by intratracheal injection at dosages from 10(7) to 10(10) pfu; a fifth group received phosphate-buffered saline alone. None of the viral injections proved to be lethal. Mild to moderate peribronchiolar and perivascular infiltration by mononuclear cells and lymphocytes, with patches of pneumonitis, was the most acute toxic effect detected by histologic analysis in the two high-dose groups. Immunohistochemical analysis of the same paraffin-embedded sections showed that infectivity and level of expression of p53 in lung tissue were dose-dependent. Our results demonstrate that Ad5CMV-p53 is a replication-defective virus that yields a relatively low degree of acute toxicity in mice; these data document a safety profile encouraging for clinical trials of Ad5CMV-p53 in the therapy of lung cancer.
Zhang
W W
WW
Department of Thoracic and Cardiovascular Surgery, University of Texas M. D. Anderson Cancer Center, Houston 77030, USA.
Alemany
R
R
Wang
J
J
Koch
P E
PE
Ordonez
N G
NG
Roth
J A
JA
D000260:Adenoviruses, Human / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology; D000818:Animals; D001980:Bronchi / Q000166:cytology; D002478:Cells, Cultured; D004261:DNA Replication; D004847:Epithelial Cells; D016158:Genes, p53 / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics / Q000633:toxicity*; D006367:HeLa Cells; D006801:Humans; D008168:Lung / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D016159:Tumor Suppressor Protein p53 / Q000633:toxicity*; D014779:Virus Replication
D016159:Tumor Suppressor Protein p53