Carbon nanotubes significantly enhance the biological activity of CpG ODN in chickens

Synthetic unmethylated cytidine-phosphate-guanosine oligodeoxynucleotides (CpG ODN) is an effective immune stimulant in chicken. To be effective CpG dosage requirement is high. High dosage increases cost of treatment and introduces toxicity. A delivery system using multi-walled carbon nanotubes (MWCNT) is utilized in this study to aid in lowering the effective dose of the immune stimulant. CpG ODNs were attached non-covalently in different ways to multi-walled carbon nanotubes (MWCNT). We assessed and selected an appropriate linking method of CpG ODN with MWCNT followed by cellular uptake studies to establish that MWCNT-conjugated CpG ODNs were delivered better than free CpG ODNs into the cell. It was observed that MWCNT-conjugated CpG ODNs were equally effective in priming the cells in vitro at 1000-fold less concentration than free CpG ODN. In vivo studies revealed that a significantly lower dose of CpG ODN, when given subcutaneously, was enough to protect chickens from a lethal challenge of bacteria. The mechanism of immune stimulation was examined by in vivo cell recruitment and in vitro cytokine production studies. MWCNT-conjugated CpG ODNs are significantly more efficacious and less toxic than free CpG ODN to qualify as a potential immune stimulant.