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Development of CpG-oligodeoxynucleotides for effective activation of rabbit TLR9 mediated immune responses
Tsung-Hsien Chuang1, Chao-Yang Lai2, Ping-Hui Tseng3
1Immunology Research Center, National Health Research Institutes, Miaoli, Taiwan; Program in Environmental and Occupational Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
Abstract:
CpG-oligodeoxynucleotides (CpG-ODN) are potent immune stimuli being developed for use as adjuvants in different species. Toll-like receptor 9 (TLR9) is the cellular receptor for CpG-ODN in mammalian cells. The CpG-ODN with 18-24 deoxynucleotides that are in current use for human and mouse cells, however, have low activity with rabbit TLR9. Using a cell-based activation assay, we developed a type of CpG-ODN containing a GACGTT or AACGTT motif in 12 phosphorothioate-modified deoxynucleotides with potent stimulatory activity for rabbit TLR9. The developed CpG-ODN have higher activities than other developed CpG-ODN in eliciting antigen-nonspecific immune responses in rabbit splenocytes. When mixed with an NJ85 peptide derived from rabbit hemorrhagic disease virus, they had potent activities to boost an antigen-specific T cell activation and antibody production in rabbits. Compared to Freund's adjuvant, the developed CpG-ODN are capable of boosting a potent and less toxic antibody response. The results of this study suggest that both the choice of CpG-motif and its length are important factors for CpG-ODN to effectively activate rabbit TLR9 mediated immune responses.
Insights
New CpG-oligodeoxynucleotides (CpG-ODN) effectively stimulate rabbit Toll-like receptor 9 (TLR9). These optimized CpG-ODN enhance immune responses and antibody production in rabbits, offering a less toxic adjuvant alternative.
Area of Science:
- Immunology
- Veterinary Medicine
- Molecular Biology
Background:
- CpG-oligodeoxynucleotides (CpG-ODN) are immune stimulants targeting Toll-like receptor 9 (TLR9).
- Standard CpG-ODN show limited efficacy with rabbit TLR9.
- Adjuvants are crucial for enhancing vaccine efficacy.
Purpose of the Study:
- To develop novel CpG-ODN with potent activity for rabbit TLR9.
- To evaluate the efficacy of these CpG-ODN as adjuvants in rabbits.
- To compare the immune response induced by developed CpG-ODN with existing adjuvants.
Main Methods:
- Developed short (12-mer) phosphorothioate CpG-ODN with GACGTT or AACGTT motifs.
- Assessed CpG-ODN activity using cell-based activation assays with rabbit splenocytes.
- Evaluated antigen-specific immune responses (T cell activation, antibody production) in rabbits immunized with NJ85 peptide and CpG-ODN.
Main Results:
- Developed 12-mer CpG-ODN demonstrated potent stimulatory activity for rabbit TLR9.
- These CpG-ODN elicited stronger antigen-nonspecific immune responses in rabbit splenocytes compared to other CpG-ODN.
- CpG-ODN adjuvants boosted antigen-specific T cell activation and antibody production against rabbit hemorrhagic disease virus peptide.
- The developed CpG-ODN induced a potent antibody response with reduced toxicity compared to Freund's adjuvant.
Conclusions:
- CpG-motif sequence and length are critical for effective rabbit TLR9 activation.
- Optimized CpG-ODN represent a promising, less toxic adjuvant for enhancing rabbit immune responses.
- This study provides a foundation for developing effective CpG-ODN-based vaccines in rabbits.
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