Adenoviral transduction of human osteoblastic cell cultures: a new perspective for gene therapy of bone diseases.
This article confirms the susceptibility of osteoblastic cells to adenoviral transduction. Osteoblasts were harvested from human cancellous bone. Cells were transduced, using various amounts of adenoviral vectors carrying the cDNA encoding interleukin-1 receptor antagonist (IL-1 Ra), or the marker genes beta-galactosidase and luciferase. Expression of the transgenes and the biological activity of IL-1 Ra produced by gene transfer were measured quantitatively in a time-course by ELISA. The rate of transduction was 100% after exposure to 1 x 10(7) infective particles of adeno-LacZ. No expression of IL-1Ra was seen after transduction with adeno-IL-1Ra at titers of 1 x 10(4) and less. However, after transduction at titers of 1 x 10(7), infective particles cells expressed IL-1 Ra consistently for 72 days, with levels up to 1 microg IL-1 Ra/1 x 10(6) cells/ 48 hours. None of the control samples expressed detectable levels of IL-1 Ra. The biological activity of the transgenic IL-1 Ra was demonstrated by its ability to suppress successfully IL-1-induced nitric oxide synthesis by rabbit articular chondrocytes. After transduction with 1 x 10(7) infective particles of the adenoluciferase vector, up to 81,000 Units transgenic luciferase/x 10(6) osteoblastic cells were measured 2 days after gene transfer. Our results show that adenovirus transduces osteoblastic cells at a high rate in vitro.
Baltzer
A W
AW
Orthopädische Universitätsklinik, Heinrich-Heine-University, Düsseldorf, Germany. axel.baltzer@uni-duesseldorf.de
Whalen
J D
JD
Stefanovic-Racic
M
M
Ziran
B
B
Robbins
P D
PD
Evans
C H
CH
D000260:Adenoviruses, Human / Q000235:genetics*; D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D001847:Bone Diseases / Q000276:immunology / Q000628:therapy*; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics / Q000627:therapeutic use*; D004279:DNA, Viral / Q000235:genetics / Q000627:therapeutic use*; D015870:Gene Expression / Q000235:genetics; D018014:Gene Transfer Techniques; D005819:Genetic Markers / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics / Q000627:therapeutic use*; D006801:Humans; D053590:Interleukin 1 Receptor Antagonist Protein; D007763:Lac Operon / Q000235:genetics; D008156:Luciferases / Q000032:analysis / Q000235:genetics*; D008875:Middle Aged; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D010006:Osteoblasts / Q000502:physiology*; D011817:Rabbits; D012795:Sialoglycoproteins / Q000032:analysis / Q000235:genetics* / Q000627:therapeutic use*; D013997:Time Factors; D014161:Transduction, Genetic / Q000235:genetics*; D019076:Transgenes / Q000235:genetics; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics*
D018076:DNA, Complementary; D004279:DNA, Viral; D005819:Genetic Markers; C507763:IL1RN protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D012795:Sialoglycoproteins; D008156:Luciferases; D019001:Nitric Oxide Synthase; D000469:Alkaline Phosphatase; D001616:beta-Galactosidase