Structural and functional characterisation of the Toll like receptor 9 of Aotus nancymaae, a non-human primate model for malaria vaccine development.
In the absence of suitable rodent animal models for Plasmodium falciparum malaria, the efficacy testing of asexual blood-stage vaccine candidates in Aotus nancymaae represents a tool to select between different formulations before conducting expensive human clinical trials. CpG oligonucleotides (ODN) specifically promote the production of pro-inflammatory and Th1-type cytokines and they enhance the immunogenicity of co-administered antigens. Toll like receptor 9 (TLR-9) binds directly and sequence-specifically to single-stranded un-methylated CpG-DNA mediating the biological effects of CpG ODN. We cloned and functionally characterised the TLR-9 cDNA of A. nancymaae. The cDNA encompassed 3,099 bp predicted to code for 1,032 amino acid residues. Results of homology searches to human TLR-9 suggested that the receptor is 93 and 94% identical at the nucleotide and amino acid sequence levels, respectively. Stimulation of splenocytes of A. nancymaae with CpG ODN resulted in proliferative responses in all animals analysed. FACS analysis of cultures incubated with CpG ODN 2006 indicated that the B cell marker CD20 was up-regulated consistent with B cell activation. The high level of sequence conservation of Aona-TLR-9 reinforces the suitability of A. nancymaae as animal model for malaria subunit vaccine development.
Spirig
Rolf
R
Department of Medical Parasitology and Biology of Infection, Molecular Immunology, Swiss Tropical Institute, Socinstrasse 57, 4002 Basel, Switzerland.
Peduzzi
Elisabetta
E
Patarroyo
Manuel E
ME
Pluschke
Gerd
G
Daubenberger
Claudia A
CA
D000595:Amino Acid Sequence; D000818:Animals; D016645:Aotidae / Q000235:genetics* / Q000276:immunology*; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D007155:Immunologic Factors / Q000008:administration & dosage / Q000378:metabolism; D066298:In Vitro Techniques; D008213:Lymphocyte Activation / Q000187:drug effects; D017780:Malaria Vaccines / Q000008:administration & dosage / Q000494:pharmacology*; D008562:Membrane Glycoproteins / Q000235:genetics; D023421:Models, Animal; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology; D010802:Phylogeny; D010963:Plasmodium falciparum / Q000276:immunology; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013154:Spleen / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D051217:Toll-Like Receptor 9; D051193:Toll-Like Receptors
C408982:CPG-oligonucleotide; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D007155:Immunologic Factors; D017780:Malaria Vaccines; D008562:Membrane Glycoproteins; D009838:Oligodeoxyribonucleotides; D011956:Receptors, Cell Surface; C495371:TLR9 protein, human; D051217:Toll-Like Receptor 9; D051193:Toll-Like Receptors