Transgene expression of biological active recombinant human granulocyte-colony stimulating factor (hG-CSF) into mouse urine.
We have generated transgenic mice that expressed human granulocyte-colony stimulating factor (hG-CSF) in their urine. In particular, the expression plasmid DNA containing mouse uroplakin II promoter was used to direct the uroepithelium-specific transcription of the transgene. In this study, the hG-CSF transcript was detected only in bladder, as was determined by RT-PCR analysis. Furthermore, hG-CSF protein was detected in the suprabasal layer of the uroepithelium and ureter, as was demonstrated by immunohistochemistry. The hG-CSF was secreted into urine at a high level (approx. 500 pg/ml), and it was able to enhance the proliferation of DMSO treated HL-60 cells, suggesting that the transgenic urine-derived hG-CSF was bioactive. However, the recombinant hG-CSF was leaked to peripheral circulation system. To examine the relationship between hG-CSF in the blood stream and the proliferation of hematopoietic cells, we tested the transgenic mouse blood with hematocrit analysis. An increase of the total number of neutrophils in the transgenic mice peripheral blood was not observed; therefore, the leakage of human G-CSF can probably be expected to do no harm to the transgenic mouse. Our results demonstrate that bladder can be safely used as a bioreactor to produce biologically important substances such as recombinant G-CSF.
Kim
Myoung Ok
MO
School of Lifesciences and Biotechnology, College of Natural Sciences, Kyungpook National University, Daegu, 702-701, Korea.
Kim
Sung Hyun
SH
Lee
Sang Ryel
SR
Kim
Kil Soo
KS
Min
Kwan Sik
KS
Lee
Hoon Taek
HT
Kim
Sun Jung
SJ
Ryoo
Zae Young
ZY
D000818:Animals; D001772:Blood Cell Count; D049109:Cell Proliferation / Q000187:drug effects; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D004622:Embryo, Mammalian / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000494:pharmacology / Q000652:urine*; D018922:HL-60 Cells; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008845:Microinjections; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019076:Transgenes / Q000235:genetics*; D060170:Uroplakin II
D008565:Membrane Proteins; D011994:Recombinant Proteins; C554658:UPK2 protein, human; C554659:Upk2 protein, mouse; D060170:Uroplakin II; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D004247:DNA