Free Feeding of CpG-Oligodeoxynucleotide Particles Prophylactically Attenuates Allergic Airway Inflammation and Hyperresponsiveness in Mice

CpG-oligodeoxynucleotides (CpG-ODNs) constitute an attractive alternative for asthma treatment. However, very little evidence is available from studies on the oral administration of CpG-ODNs in animals. Previously, we developed acid-resistant particles (named ODNcap) as an oral delivery device for ODNs. Here, we showed that free feeding of an ODNcap-containing feed prophylactically attenuates allergic airway inflammation, hyperresponsiveness, and goblet cell hyperplasia in an ovalbumin-induced asthma model. Using transcriptomics-driven approaches, we demonstrated that injury of pulmonary vein cardiomyocytes accompanies allergen inhalation challenge, but is inhibited by ODNcap feeding. We also showed the participation of an airway antimicrobial peptide (Reg3γ) and fecal microbiota in the ODNcap-mediated effects. Collectively, our findings suggest that daily oral ingestion of ODNcap may provide preventive effects on allergic bronchopulmonary insults via regulation of mechanisms involved in the gut-lung connection.

In ovo Feeding as a Tool for Improving Performance and Gut Health of Poultry: A Review

Early growth and development of the gastrointestinal tract are of critical importance to enhance nutrients' utilization and optimize the growth of poultry. In the current production system, chicks do not have access to feed for about 48–72 h during transportation between hatchery and production farms. This lag time affects early nutrient intake, natural exposure to the microbiome, and the initiation of beneficial stimulation of the immune system of chicks. In ovo feeding can provide early nutrients and additives to embryos, stimulate gut microflora, and mitigate the adverse effects of starvation during pre-and post-hatch periods. Depending on the interests, the compounds are delivered to the embryo either around day 12 or 17 to 18 of incubation and via air sac or amnion. In ovo applications of bioactive compounds like vaccines, nutrients, antibiotics, prebiotics, probiotics, synbiotics, creatine, follistatin, L-carnitine, CpG oligodeoxynucleotide, growth hormone, polyclonal antimyostatin antibody, peptide YY, and insulin-like growth factor-1 have been studied. These compounds affect hatchability, body weight at hatch, physiological functions, immune responses, gut morphology, gut microbiome, production performance, and overall health of birds. However, the route, dose, method, and time of in ovo injection and host factors can cause variation, and thereby inconsistencies in results. Studies using this method have manifested the benefits of injection of different single bioactive compounds. But for excelling in poultry production, researchers should precisely know the proper route and time of injection, optimum dose, and effective combination of different compounds. This review paper will provide an insight into current practices and available findings related to in ovo feeding on performance and health parameters of poultry, along with challenges and future perspectives of this technique.

A Novel CpG Oligodeoxynucleotide Enhanced the Therapeutic Effect of Epirubicin on Bladder Tumors in Rats

Background CpG oligodeoxynucleotides, which boast anti-inflammatory, anti-infectious, and chemotherapeutic activities, are promising immunomodulators. Recently, some preclinical studies have highlighted the potent immunostimulatory and anti-tumor effects of CpG oligodeoxynucleotides, which has aroused interest in their potential clinical effects on human cancers. Methods In this study, we evaluated the therapeutic effect of a new type of CpG oligodeoxynucleotide whose sequence (5’-AACGTTGTCGTCGACGTCGTCGTCAGGCCTGACGTTATCGATGGCGTTGTCGTCAACGTTGTCGTTAACGTT-3’) was designed by our laboratory in combination with epirubicin in a bladder tumor rat model induced by N-methyl-N-nitrosourea instillation. Moreover, we explored the safety of the novel CpG oligodeoxynucleotide for bladder tumor therapy by observing the degree of cystolith in the bladder and comparing the results against those of Bacillus Calmette–Guérin therapy, which is a gold-standard treatment for bladder tumor. Results All results showed that CpG oligodeoxynucleotide combined with epirubicin significantly inhibited the growth of bladder tumors and reduced the pathological grading. As compared with bladder cells or cytokines observed under the positive control or epirubicin-alone treatment conditions, all indexes including histopathological grading, Mutation P53 gene protein expression, and Interleukin-2 (IL-2) level were significantly optimized by instillation of CpG oligodeoxynucleotide. Immunohistochemical examination indicated that CpG oligodeoxynucleotide reduced the expression of Mutation P53 gene protein in the bladder tumor rat model. Specifically, the level of IL-2 in rat serum was increased by more than 30% CpG oligodeoxynucleotide treatment combined with epirubicin. Also, in comparison with the degree of cystolith observed in the Bacillus Calmette–Guérin group, no obvious side effects were caused by CpG oligodeoxynucleotide. Conclusions CpG oligodeoxynucleotide as an immunomodulator can enhance the efficacy of epirubicin and presents higher safety than Bacillus Calmette–Guérin in treating bladder cancer.

Sharpening up tumor microenvironment to enhance the efficacy of immune checkpoint blockade on head and neck cancer using a CpG-oligodeoxynucleotide

Head and neck cancers are a type of life-threatening cancers characterized by an immunosuppressive tumor microenvironment. Only less than 20% of the patients respond to immune checkpoint blockade therapy, indicating the need for a strategy to increase the efficacy of immunotherapy for this type of cancers. Previously, we identified a type B CpG-oligodeoxynucleotide (CpG-ODN) called CpG-2722, which has the universal activity of eliciting an immune response in grouper, mouse, and human cells. In this study, we further characterized and compared its cytokine-inducing profiles with different types of CpG-ODNs. The antitumor effect of CpG-2722 was further investigated alone and in combination with an immune checkpoint inhibitor in a newly developed syngeneic orthotopic head and neck cancer animal model. Along with other inflammatory cytokines, CpG-2722 induces the gene expressions of interleukin-12 and different types of interferons, which are critical for the antitumor response. Both CpG-2722 and anti-programmed death (PD)-1 alone suppressed tumor growth. Their tumor suppression efficacies were further enhanced when CpG-2722 and anti-PD-1 were used in combination. Mechanistically, CpG-2722 shaped a tumor microenvironment that is favorable for the action of anti-PD-1, which included promoting the expression of different cytokines such as IL-12, IFN-β, and IFN-γ, and increasing the presence of plasmacytoid dendritic cells, M1 macrophages, and CD8 positive T cells. Overall, CpG-2722 provided a priming effect for CD8 positive T cells by sharpening the tumor microenvironment, whereas anti-PD-1 released the brake for their tumor-killing effect, resulting in an enhanced efficacy of the combined CpG-2722 and anti-PD-1.

Immunopotentiation by Lymph-Node Targeting of a Malaria Transmission-Blocking Nanovaccine

A successful malaria transmission blocking vaccine (TBV) requires the induction of a high antibody titer that leads to abrogation of parasite traversal of the mosquito midgut following ingestion of an infectious bloodmeal, thereby blocking the cascade of secondary human infections. Previously, we developed an optimized construct UF6b that elicits an antigen-specific antibody response to a neutralizing epitope of Anopheline alanyl aminopeptidase N (AnAPN1), an evolutionarily conserved pan-malaria mosquito midgut-based TBV target, as well as established a size-controlled lymph node targeting biodegradable nanoparticle delivery system that leads to efficient and durable antigen-specific antibody responses using the model antigen ovalbumin. Herein, we demonstrate that co-delivery of UF6b with the adjuvant CpG oligodeoxynucleotide immunostimulatory sequence (ODN ISS) 1018 using this biodegradable nanoparticle vaccine delivery system generates an AnAPN1-specific immune response that blocks parasite transmission in a standard membrane feeding assay. Importantly, this platform allows for antigen dose-sparing, wherein lower antigen payloads elicit higher-quality antibodies, therefore less antigen-specific IgG is needed for potent transmission-reducing activity. By targeting lymph nodes directly, the resulting immunopotentiation of AnAPN1 suggests that the de facto assumption that high antibody titers are needed for a TBV to be successful needs to be re-examined. This nanovaccine formulation is stable at -20°C storage for at least 3 months, an important consideration for vaccine transport and distribution in regions with poor healthcare infrastructure. Together, these data support further development of this nanovaccine platform for malaria TBVs.