A novel and rapid prediction assay for the effectiveness of IL-6 receptor specific antisense oligonucleotides by proliferation inhibition of an interleukin-6 dependent cell line.
Interleukin-6 (IL-6) belongs to a family of cytokines that use receptors consisting of a common signal-transducing chain (gp130). Baf/3 cells transfected with the human IL-6 receptor (IL-6R) and gp130 (Baf/3-gp130/IL-6R) can only grow in medium containing IL-6. We attempted to interrupt the signal transducing pathway of IL-6 with the help of antisense oligonucleotides (ASOs) designed against the IL-6R. We used 18 different kinds of antisense oligonucleotides of overlapping sequences around the translational start codon of the human IL-6R. Sense ASOs were used as a control. The proliferation of cells was analysed by H-thymidine incorporation. Cell surface expression of the IL-6R was assessed by FACS analysis. We identified three ASOs which strongly inhibited the proliferation of IL-6 dependent transfected Baf/3 cells. Flow cytometric studies on the suppression of surface expression of IL-6R by ASOs showed a similar pattern. These results should help to clarify the structural requirements of functionally effective ASOs in the inhibition of IL-6R.
Lia Varga
V
V
Department of Genetics, Cell- and Immunobiology, Semmelweis University of Medicine, Budapest, Hungary.
Falus
A
A
Pallinger
E
E
Novák
I
I
Fülöp
K A
KA
Toth
S
S
Rose-John
S
S
D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D005434:Flow Cytometry; D006801:Humans; D015850:Interleukin-6 / Q000494:pharmacology*; D016376:Oligonucleotides, Antisense / Q000235:genetics* / Q000494:pharmacology; D019947:Receptors, Interleukin-6 / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D013379:Substrate Specificity; D013997:Time Factors
D015850:Interleukin-6; D016376:Oligonucleotides, Antisense; D019947:Receptors, Interleukin-6