Immunoregulatory activities of nanoparticle-forming oligodeoxynucleotides

Abstract

Innate immune system is activated by a wide range of microbial by products leading to an immediate immune activation primarily designed to neutralize and control the invading insult. The cells of the innate immune system also instruct the development of antigen-specific adaptive immunity. While TLR9 is triggered by bacterial DNA, extended and over-exuberant immune response poses a threat since it may exacerbate cell and tissue destruction leading to organ failure. Telomeric TTAGGG conserved motifs are previously reported to antagonize TLR mediated events. The down-regulatory effect of these motifs may help to restore the desired homeostatic balance of the immune system. While CpG ODN patterned after bacterial DNA can be harnessed in different clinical settings to provide an advantage to host to resist infectious diseases, control tumor growth or alleviate allergic symptoms, the immunosuppressive telomeric motifs could be effectively applied in controlling systemic anti-inflammatory or autoimmune related disorders. Several challenges exist in the utilization of synthetic ODNs in the clinic. The first challenge is that conventional classes of synthetic ODNs exhibit different properties. K-type ODNs are more effective in proliferation and activation of B cells and DC. D-type ODNs are in nanoparticle forms, lead to anti-viral type I IFN production and mature monocytes into DCs. Of note, the efficacy of these synthetic ODNs is reduced under physiological conditions due to premature clearance and low levels of internalization. Moreover, D-ODNs as one of the most potent IFNα inducing TLR9 ligands possess a large-scale production problem due to 3’polyGruns, which hamper their entry into the clinic. We have designed a novel class of ODN, designated as ODN420, devoid of polyGs that can undergo nanoparticle formation necessary for its IFNα induction. Ex vivo stimulation of mouse splenocytes and in vivo administration of ODN420 have revealed that this ODN exhibits higher immunostimulatory potential and is more stable than most commonly used ODNs due to its nanoparticle-forming ability. Another interesting finding is that ODN420 with the natural phosphodiester (PO) backbone is at least as potent as its more stable counterpart with the modified phosphorothioate backbone. Furthermore, it combines superior properties of conventional classes of K and D-ODNs. These results have been reproduced in human peripheral blood mononuclear cells by various assays. Next, we have analyzed whether this ODN could be utilized as a vaccine adjuvant and an anti-cancer agent with two independent experiments. Our immunization results demonstrate that ODN420 induces a higher level of Th1-mediated response than conventional ODNs and is a promising candidate as a vaccine adjuvant. This response is hampered when ODN420 is used in combination with ODN-A151. In the tumor xenograft model, ODN420 has promoted partial remission of the tumors or delayed the tumor growth. This knowledge will pave the way for more effective immunotherapeutic approaches.