W. Denham et al., TRANSIENT HUMAN GENE-THERAPY - A NOVEL CYTOKINE REGULATORY STRATEGY FOR EXPERIMENTAL PANCREATITIS, Annals of surgery, 227(6), 1998, pp. 812-819
Objective The purpose of this study was to evaluate the ability to tra
nsfect a murine pancreas with a human cytokine regulatory gene (interl
eukin-10 [1L-10]) and examine the duration of transgene expression, it
s effect on the normal pancreas, and its antiinflammatory effect durin
g acute pancreatitis. Summary Background Data Interleukin-1 beta and t
umor necrosis factor-alpha are known detrimental mediators during the
progression of acute pancreatitis, and blockade of either cytokine res
ults in decreased severity of pancreatitis and improved survival. Alth
ough gene therapy has been proposed as a method to deliver protein-bas
ed therapy during a number of conditions, no means of effectively tran
sfecting the pancreas without inducing injury has been developed. Meth
ods A plasmid-human IL-10 construct (pMP6-hlL-10) complexed with catio
nic liposomes was administered by single intrapenitoneal injection to
healthy mice. Effective transfection (reverse transcriptase-polymerase
chain reaction for hIL-10 mRNA), transfected cell type (in situ polym
erase chain reaction for hIL-10 DNA), and the effect on the normal pan
creas were determined. Additional animals were transfected to determin
e the effects of this regulatory gene on the severity of pancreatitis.
Results Nearly 80% of all pancreatic cells expressed human DNA that w
as subsequently transcribed into mRNA through day 14. The transfection
event had no effect on amylase, lipase, or pancreatic histologic appe
arance. Successful transfection could attenuate subsequently induced p
ancreatitis (all parameters p < 0.05). Conclusions Transient transfect
ion of a human IL-10 gene can be accomplished into all cell types of m
urine pancreata using a plasmid/liposome vector, The DNA is effectivel
y transcribed into intact mRNA and does not cause inflammation or acin
ar cell damage. Transfer of this cytokine regulatory gene decreases th
e severity of pancreatitis, demonstrating a benefit of gene therapy du
ring this acute inflammatory process.