Adenovirus-transduced human butyrylcholinesterase in mouse blood functions as a bioscavenger of chemical warfare nerve agents.
Human serum butyrylcholinesterase (Hu BChE) is a promising therapeutic against the toxicity of chemical warfare nerve agents. We have showed previously that recombinant (r) Hu BChE can be expressed at very high levels, 400 to 600 U/ml in mouse blood, by delivering the Hu BChE gene using adenovirus (Ad). Here, we report the biochemical properties of the Ad-expressed full-length and truncated rHu BChE in mouse blood. The molecular sizes of the full-length rHu BChE subunit and its oligomers were similar to those of native Hu BChE, although only a small portion of the full-length rHu BChE subunit underwent assembly into dimers and tetramers. As expected, Ad containing the truncated Hu BChE gene transduced the expression of monomeric rHu BChE only. Compared with 415 U of rHu BChE per milliliter in blood, tissues including liver, lung, heart, brain, kidney, muscle, intestine, diaphragm, salivary gland, and fat expressed <10 U/g of rHu BChE activity. Ad-expressed rHu BChE in mouse blood neutralized soman and O-ethyl S-2-N,N-diisopropylaminoethyl methylphosphonothiolate at rates similar to those of native Hu BChE and rHu BChE expressed in vitro. Because the expression of rHu BChE rapidly decreased 6 days after virus administration, sera were assayed for the presence of anti-Hu BChE antibodies. Anti-Hu BChE antibodies were detected on day 7 and in increased amounts thereafter, which coincided with the loss of Hu BChE expression in sera. In conclusion, the delivery of Hu BChE gene using Ad can be a promising strategy that can provide protection against multiple lethal doses of chemical warfare nerve agents in vivo.
Chilukuri
Nageswararao
N
Department of Molecular Pharmacology, Division of Biochemistry, Walter Reed Army Institute of Research, Silver Spring, MD 20910-7500, USA. nageswararao.chilukuri@us.army.mil
Duysen
Ellen G
EG
Parikh
Kalpana
K
diTargiani
Robert
R
Doctor
Bhupendra P
BP
Lockridge
Oksana
O
Saxena
Ashima
A
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002091:Butyrylcholinesterase / Q000097:blood* / Q000235:genetics* / Q000378:metabolism; D002619:Chemical Warfare Agents / Q000378:metabolism*; D002800:Cholinesterase Inhibitors / Q000378:metabolism*; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D011994:Recombinant Proteins / Q000097:blood / Q000235:genetics / Q000378:metabolism; D012999:Soman / Q000378:metabolism*; D014018:Tissue Distribution; D014161:Transduction, Genetic
D002619:Chemical Warfare Agents; D002800:Cholinesterase Inhibitors; D011994:Recombinant Proteins; D012999:Soman; D002091:Butyrylcholinesterase