Conjugates of Double-Stranded Oligonucleotides with Poly(ethylene glycol) and Keyhole Limpet Hemocyanin: A Model for Treating Systemic Lupus Erythematosus

• Published in: Bioconjugate chemistry Vol. 5; no. 5; pp. 390 - 399
• Main Authors: Jones, David S, Hachmann, John P, Osgood, Stephen A, Hayag, Merle S, Barstad, Paul A, Iverson, G. Michael, Coutts, Stephen M
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01-09-1994
• Subjects: Alkylation; Animals; Antibody Formation; Antigens - immunology; B-Lymphocytes - immunology; Electrophoresis, Polyacrylamide Gel; Female; Hemocyanins - chemistry; Immune Tolerance; Immunization; In Situ Hybridization; Lupus Erythematosus, Systemic - therapy; Magnetic Resonance Spectroscopy; Mice; Mice, Inbred C57BL; Oligonucleotides - chemistry; Oligonucleotides - immunology; Oligonucleotides - therapeutic use; Polyethylene Glycols - chemistry; Spectrophotometry, Ultraviolet
• ISSN: 1043-1802; 1520-4812
• DOI: 10.1021/bc00029a003

Two types of oligonucleotides were synthesized with linker groups attached at the 5'-end. Both were repeating dimers of deoxyribocytidine and deoxyriboadenosine. A 20-mer was prepared with a thiol-containing linker, masked as a disulfide, and a 50-mer was prepared with a vicinal diol-containing linker. A tetraiodoacetylated poly(ethylene glycol) (PEG) derivative was synthesized and reacted with the thiol-containing 20-mer to provide an oligonucleotide PEG conjugate of precisely four oligonucleotides on each PEG carrier. The vicinal diol on the 50-mer was oxidized to an aldehyde and conjugated to keyhole limpet hemocyanin (KLH) to provide an oligonucleotide-KLH conjugate by reductive alkylation. The conjugates were annealed with complementary (TG)n strands. While the double-stranded oligonucleotide-KLH conjugate is an immunogen, eliciting the synthesis of antibodies against oligonucleotides, the PEG conjugate has the biological property of specifically suppressing (tolerizing) B cells which make antibodies against the immunizing oligonucleotide.