Expression of recombinant human granulocyte macrophage-colony stimulating factor (hGM-CSF) in mouse urine.
We have generated transgenic mice expressing human granulocyte macrophage-colony stimulating factor (hGM-CSF) in urine. In particular, the expression plasmid DNA containing mouse uroplakin II promoter was used to direct uroepithelium-specific transcription of transgene. In this study, hGM-CSF transcript was detected only in bladder uroepithelium as determined by northern blot analysis. Furthermore, hGM-CSF protein was detected in the suprabasal layer of the uroepithelium and ureter by immunohistochemistry. The hGM-CSF was secreted into urine at high level (up to 180 ng/ml), and enhanced proliferation of hGM-CSF-dependent human acute monocyte leukemic cells, suggesting that transgenic urine-derived hGM-CSF was bioactive. This is the first case of demonstrating biological activity of a cytokine produced in the urine of a transgenic animal. Our results demonstrate that bladder can be used as a bioreactor to produce biologically important substances. In addition, it suggests a potential application of bladder expression system to livestock for high-yield production of pharmaceuticals.
Ryoo
Z Y
ZY
Department of Immunobiology and Dermatology, College of Medicine, Catholic University of Korea, Seoul.
Kim
M O
MO
Kim
K E
KE
Bahk
Y Y
YY
Lee
J W
JW
Park
S H
SH
Kim
J H
JH
Byun
S J
SJ
Hwang
H Y
HY
Youn
J
J
Kim
T Y
TY
D000818:Animals; D001483:Base Sequence; D001709:Biotechnology / Q000379:methods*; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D004848:Epithelium / Q000378:metabolism; D015870:Gene Expression; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology / Q000652:urine*; D006801:Humans; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D009928:Organ Specificity; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins; D019076:Transgenes / Q000235:genetics; D001743:Urinary Bladder / Q000378:metabolism; D060170:Uroplakin II
D008565:Membrane Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; C554658:UPK2 protein, human; C554659:Upk2 protein, mouse; D060170:Uroplakin II; D016178:Granulocyte-Macrophage Colony-Stimulating Factor