BHQ-0 CPG; Glycolate, 500 Å

CpG islands are generally known as the epigenetic regulatory regions in accordance with histone modifications, methylation, and promoter activity. There is a significant need for the exact mapping of DNA methylation in CpG islands to understand the diverse biological functions. However, the precise identification of CpG islands from the whole genome through experimental and computational approaches is still challenging. Numerous computational methods are being developed to detect the CpG-enriched regions, effectively, to reduce the time and cost of the experiments. Here, we review some of the latest computational CpG detection methods that utilize clustering, patterns and physical-distance like parameters for CpG island detection. The comparative analyses of the methods relying on different principles and parameters allow prioritizing the algorithms for specific CpG associated datasets to achieve higher accuracy and sensitivity. A number of computational tools based on the window, Hidden Markov Model, density and distance-/length-based algorithms are being applied on human or mammalian genomes for accurate CpG detection. Comparative analyses of CpG island detection algorithms facilitate to prefer the method according to the target genome and required parameters to attain higher accuracy, specificity, and performance. There is still a need for efficient computational CpG detection methods with lower false-positive results. This review provides a better understanding about the principles of tools that will assist to prioritize and develop the algorithms for accurate CpG islands detection.

Toll-like receptors (TLRs) are pattern-recognition receptors that detect a wide variety of microbial pathogens for the initiation of host defense immunological responses. Thirteen TLRs have been identified in mammals, and teleosts contain 22 mammalian or non-mammalian TLRs. Of these, TLR9 and TLR21 are the cytosine-phosphate-guanosine-oligodeoxynucleotides (CpG-ODNs) recognition TLRs in teleosts. TLR9 is a mammalian TLR expressed in teleost but not in the avian species. TLR21 is a non-mammalian TLR expressed in both teleost and the avian species. Synthetic CpG-ODNs are potent immunostimulants that are being studied for their application against tumors, allergies, and infectious diseases, and as a vaccine adjuvant in humans. The immunostimulatory effects of CpG-ODNs as vaccine adjuvants and their antimicrobial function in domestic animals and teleosts are also being investigated. Most of our current knowledge about the molecular basis for the immunostimulatory activity of CpG-ODNs comes from earlier studies of the interaction between CpG-ODN and TLR9. More recent studies indicate that in addition to TLR9, TLR21 is another receptor for CpG-ODN recognition in teleosts to initiate immune responses. Whether these two receptors have differential functions in mediating the immunostimulatory activity of CpG-ODN in teleost has not been well-studied. Nevertheless, the existence of two recognition TLRs suggests that the molecular basis for the immunostimulatory activity of CpG-ODN in teleosts is different and more complex than in mammals. This article reviews the current knowledge of TLR9 and TLR21 activation by CpG-ODNs. The key points that need to be considered for CpG-ODNs as immunostimulants with maximum effectiveness in activation of immune responses in teleosts are discussed. This includes the structure/activity relationship of CpG-ODN activities for TLR9 and TLR21, the structure/functional relationship of these two TLRs, and differential expression levels and tissue distributions for these two TLRs.

CpG oligodeoxynucleotides (ODNs) have attracted increasing attention as immunotherapeutic agents. However, efficient transfection of CpG ODNs into the immune cells remains a big challenge. In this study, for the first time, we reported that silk fibroin (SF) could function as an efficient carrier for CpG ODNs. A novel strategy was developed to prepare SF-CpG ODNs nanoparticles (NPs) based on self-assembly of SF. The as-prepared SF-CpG NPs were spherical in shape and were uniformly dispersed. SF-CpG NPs exhibited good stability and biocompatibility. SF-CpG NPs possessed significantly enhanced (7 folds) cellular uptake compared with CpG ODNs. Release of CpG ODNs from SF-CpG NPs was accelerated in environment-mimicking TLR9-localized endo/lysosome. SF-CpG NPs stimulated about four folds higher levels of immune cytokines and nitric oxide compared with CpG ODNs. Our results suggested that SF notably improved the CpG ODNs delivery. SF-CpG NPs have strong potential in immunotherapy.