Development of CpG-oligodeoxynucleotides for effective activation of rabbit TLR9 mediated immune responses

Tsung-Hsien Chuang; Chao-Yang Lai; Ping-Hui Tseng; Chiun-Jye Yuan; Li-Chung Hsu

doi:10.1371/journal.pone.0108808

Development of CpG-oligodeoxynucleotides for effective activation of rabbit TLR9 mediated immune responses

PLoS One. 2014 Sep 30;9(9):e108808. doi: 10.1371/journal.pone.0108808. eCollection 2014.

Authors

Tsung-Hsien Chuang¹, Chao-Yang Lai², Ping-Hui Tseng³, Chiun-Jye Yuan⁴, Li-Chung Hsu⁵

Affiliations

¹ Immunology Research Center, National Health Research Institutes, Miaoli, Taiwan; Program in Environmental and Occupational Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
² Immunology Research Center, National Health Research Institutes, Miaoli, Taiwan.
³ Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei, Taiwan.
⁴ Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan.
⁵ Institute of Molecular Medicine, National Taiwan University, Taipei, 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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adjuvants, Immunologic / chemistry
Adjuvants, Immunologic / pharmacology*
Animals
Antibody Formation / drug effects
Base Sequence
Cytokines / metabolism
HEK293 Cells
Hemorrhagic Disease Virus, Rabbit / metabolism
Humans
Immunoglobulin M / metabolism
Lymphocyte Activation / drug effects*
Oligodeoxyribonucleotides / chemistry
Oligodeoxyribonucleotides / pharmacology*
Peptides / chemistry
Peptides / pharmacology
Phosphates / chemistry
Rabbits
Spleen / drug effects
Spleen / metabolism
T-Lymphocytes / immunology
T-Lymphocytes / metabolism
Toll-Like Receptor 9 / metabolism*

Substances

Adjuvants, Immunologic
CPG-oligonucleotide
Cytokines
Immunoglobulin M
Oligodeoxyribonucleotides
Peptides
Phosphates
Toll-Like Receptor 9
phosphorodithioic acid

Grants and funding

This work was supported in part by grants from the National Health Research Institutes, Taiwan to THC (IM-102-PP-02), and from the National Science Council of Taiwan to THC (NSC102-2320-B-400-009-MY3) and LCH (NSC102-2325-B-002-06). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.