[Tumor-selective replication, cytotoxicity and GM-CSF production of oncolytic recombinant adenovirus in KH901 injection].
OBJECTIVE
To study the tumor-selective replication, cytotoxicity and GM-CSF production of the recombinant virus in KH901 injection used to infect the cells cultured in vitro.


METHODS
A panel of tumor and normal cells was infected with recombinant adenovirus in KH901 and wild-type adenovirus type 5 at a MOI of 2 PPC, the cells were harvested at 72 hours after infection and made a titer after three cycles of freeze/thaw; A panel of tumor and normal cells was infected with recombinant adenovirus KH901 at MOI of 1 or 10 PPC. For 24 hours after infection the medium was harvested to determine the biological activity of GM-CSF; A panel of tumor and normal cells was infected with KH901 of recombinant adenovirus and wild-type adenovirus type 5 at MOIs of 0, 0.1, 1, 10, 100, and 1000 PPC. At 7 days after infection, cell viability was determined by the MTT assay, and ECso was determined too.


RESULTS
The data showed that wild-type adenovirus type 5 replicated efficiently in and killed both the tumor and normal cells, however, the recombinant adenovirus in KH901 replicated hugely in tumor cells [(2526.4+/-136.8)-(2796.6+/-104.6) TCID50/cell), and produced significant amount of GM-CSF [(1177. 793 +/-6.62)-(3924.497+/-17.79) IU/(10(6) cell x 24h)] and killed the tumor cells [EC50: (0.31+/-0.06)-(0.19+/- 0.01) pfu/cell] while was replicating poorly in non-permissive human normal cells [(56.8+/-9.2)-(90.1+/-14.4) TCID50/ cell], and producing very small amount of GM-CSF [(13.397+/-0.82) IU/(10(6)cell x 24 h)] and attenuating human primary cells killed [EC50: (92.33 +/- 9.12)-(121.20 +/- 19.94) pfu/cell], with which there was statistically a significant difference between wild-type adenovirus type 5 and recombinant adenovirus in KH901 (P<0.05).


CONCLUSIONS
In vitro studies show that the tumor-selective replication, cytotoxicity, GM-CSF production of recombinant adenovirus lead the injection KH901 containing the recombinant adenovirus, as oncolytic agent, to have a potential utility for the treatment of solid tumors.
Shen
Fu-bing
FB
National Chengdu Center of Safety Evaluation of Traditional Chinese Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
Chang
Jian-hua
JH
Yang
Chun
C
Li
Jin
J
Guo
Yong
Y
Yi
Bo
B
Li
Hong-xia
HX
Ye
Xue-fei
XF
Wang
Li
L
D000256:Adenoviridae / Q000235:genetics / Q000502:physiology*; D045744:Cell Line, Tumor; D004274:DNA, Recombinant / Q000235:genetics*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D007267:Injections; D009369:Neoplasms / Q000235:genetics / Q000473:pathology / Q000821:virology*; D050504:Oncolytic Viruses / Q000235:genetics / Q000502:physiology*; D019562:Viral Load; D014779:Virus Replication
D004274:DNA, Recombinant; D016178:Granulocyte-Macrophage Colony-Stimulating Factor