Comprehensive immunoprofiling and systematic adjuvant comparisons for identifying suitable vaccine: Adjuvant pairings.

Adjuvants are critical components of vaccines that enhance the host immune response to the vaccine antigen, however, only a small number of adjuvants are used in vaccines approved for human use. This is in part due to the slow process of novel adjuvants advancing from preclinical models to human studies, and modest mechanistic insights obtained using standard immunological methods to justify selection of a particular adjuvant for clinical evaluation. Here, we discuss several aspects of current adjuvant research and strategies to better assess the complex pathways triggered by adjuvant candidates that can increase adjuvanticity and vaccine efficacy while minimizing reactogenicity. We propose a more systematic use of broad immunoprofiling, coupled with data integration using computational and mathematical modeling. This comprehensive evaluation of the host immune response will facilitate the selection of the most appropriate adjuvant for a vaccine, ultimately leading to the expeditious evaluation of novel adjuvants for vaccines against emerging infectious diseases, which will prove especially valuable during a pandemic where speed is of the essence when developing vaccines.

Construction of a prognostic signature of RFC5 immune-related genes in patients with cervical cancer.

BACKGROUND: Cervical cancer (CC) is a malignant tumor threatening women's health. Replication factor C (RFC) 5 is significantly highly expressed in CC tissues, and the immune microenvironment plays a crucial role in tumor initiation, progression, and metastasis. OBJECTIVE: To determine the prognostic role of RFC5 in CC, analyze the immune genes significantly associated with RFC5, and establish a nomogram to evaluate the prognosis of patients with CC. METHODS: High RFC5 expression in patients with CC was analyzed and verified through TCGA GEO, TIMER2.0, and HPA databases. A risk score model was constructed using RFC5-related immune genes identified using R packages. Combining the risk score model and clinical information of patients with CC, a nomogram was constructed to evaluate the prognosis of patients with CC. RESULTS: Comprehensive analysis showed that the risk score was a prognostic factor for CC. The nomogram could predict the 3-year overall survival of patients with CC. CONCLUSIONS: RFC5 was validated as a biomarker for CC. The RFC5 related immune genes were used to establish a new prognostic model of CC.

Transient and tunable CRISPRa regulation of APOBEC/AID genes for targeting hepatitis B virus.

APOBEC/AID cytidine deaminases play an important role in innate immunity and antiviral defenses and were shown to suppress hepatitis B virus (HBV) replication by deaminating and destroying the major form of HBV genome, covalently closed circular DNA (cccDNA), without toxicity to the infected cells. However, developing anti-HBV therapeutics based on APOBEC/AID is complicated by the lack of tools for activating and controlling their expression. Here, we developed a CRISPR-activation-based approach (CRISPRa) to induce APOBEC/AID transient overexpression (>4-800,000-fold increase in mRNA levels). Using this new strategy, we were able to control APOBEC/AID expression and monitor their effects on HBV replication, mutation, and cellular toxicity. CRISPRa prominently reduced HBV replication (∼90%-99% decline of viral intermediates), deaminated and destroyed cccDNA, but induced mutagenesis in cancer-related genes. By coupling CRISPRa with attenuated sgRNA technology, we demonstrate that APOBEC/AID activation can be precisely controlled, eliminating off-site mutagenesis in virus-containing cells while preserving prominent antiviral activity. This study untangles the differences in the effects of physiologically expressed APOBEC/AID on HBV replication and cellular genome, provides insights into the molecular mechanisms of HBV cccDNA mutagenesis, repair, and degradation, and, finally, presents a strategy for a tunable control of APOBEC/AID expression and for suppressing HBV replication without toxicity.

Effects of immunostimulators of microbial origin on T cells of pigs vaccinated with attenuated vaccine against Aujeszky's disease.

Aujeszky's disease (AD) is a viral infectious disease caused by Suid herpesvirus 1 (SuHV-1). Vaccination and eradication of AD in domestic pigs is possible using marker vaccines with attenuated or inactivated SuHV-1, or subunit vaccines. However, vaccines with attenuated SuHV-1 have shown to be more potent in inducing strong cell-mediated immune response. The studies have shown that Parapoxvirus ovis, as well as Propionibacterium granulosum with lipopolysacharides (LPS) of Escherichia coli have pronounced immunomodulatory effects and that in combination with the vaccines can induce stronger humoral and cellular immune responses than use of vaccines alone. In our study distribution of peripheral blood T cell subpopulations was analysed after administration of vaccine alone (attenuated SuHV-1), immunostimulators (inactivated Parapoxvirus ovis or combination of an inactivated P. granulosum and detoxified LPS of E. coli) and combinations of vaccine with each immunostimulator to the 12-week old piglets. Throughout the study no significant changes were found in the proportions of γδ and most αß T cell subpopulations analysed. However, on the seventh day of the study combination of an inactivated P. granulosum and LPS of E. coli with vaccine induced transient but significant increase of the proportions of CD4+CD8α+ and CD4-CD8α+ αß T cells, that have been strongly associated with early protection of SuHV-1 infected pigs. Our findings indicate that combination of inactivated P. granulosum and detoxified E. coli LPS could be used for enhancement of a cellular immune response induced by vaccines against AD.

Microalgae in Terms of Biomedical Technology: Probiotics, Prebiotics, and Metabiotics.

Green, red, brown, and diatomic algae, as well as cyanobacteria, have been in the focus of attention of scientists and technologists for over 5 decades. This is due to their importance as efficient and economical producers of food additives, cosmetics, pharmaceuticals, biofertilizers, biofuels, and wastewater bioremediation agents. Recently, the role of microalgae has increasingly been considered in terms of their probiotic function, i.e., of their ability to normalize the functioning of the microbiota of humans and agricultural animals and to produce biologically active substances, including hormones, neurotransmitters, and immunostimulators. A separate brief subsection of the review deals with the potential functions of microalgae with respect to the brain and psyche, i.e., as psychobiotics. Moreover, algal polysaccharides and some other compounds can be broken down to short fragments that will stimulate the development of useful intestinal microorganisms, i.e., function as efficient prebiotics. Finally, many components of microalgal cells and chemical agents produced by them can exert important health-promoting effects per se, which indicates that they are as potentially valuable metabiotics (the term preferred by late Prof. B.A. Shenderov), which are alternatively denoted as postbiotics in the literature.

Pattern Recognition Receptor Ligands as an Emerging Therapeutic Agent for Latent HIV-1 Infection.

Toll-like receptors (TLRs) were first identified as molecular sensors that transduce signals from specific structural patterns derived from pathogens; their underlying molecular mechanisms of recognition and signal transduction are well-understood. To date, more than 20 pattern-recognition receptors (PRRs) have been reported in humans, some of which are membrane-bound, similar to TLRs, whereas others are cytosolic, including retinoic acid-inducible gene-I (RIG-I)-like receptors (RLRs), nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs), and stimulator of interferon genes (STING). Clinically, PRR ligands have been developed as vaccine adjuvants to activate innate immunity and enhance subsequent antigen-specific immune responses. Recently, PRR ligands have been used as direct immunostimulators to enhance immune responses against infectious diseases and cancers. HIV-1 remains one of the world's most significant public health challenges. Without the elimination of HIV-1 latently infected cells, patients require lifelong combination antiretroviral therapy (cART), while research aimed at a functional cure for HIV-1 infection continues. Based on the concept of "shock and kill," a latency-reversing agent (LRA) has been developed to reactivate latently infected cells and induce cell death. However, previous research has shown that LRAs have limited efficacy in the eradication of these reservoirs in vivo. Besides, PRR ligands with anti-retroviral drugs have been developed for use in HIV treatment for these years. This mini-review summarizes the current understanding of the role of PRR ligands in AIDS research, suggests directions for future research, and proposes potential clinical applications.

Targeting strategies of liposomal subunit vaccine delivery systems to improve vaccine efficacy.

Liposomes are versatile delivery systems and immunological adjuvants that not only can load various antigens, such as proteins, peptides, nucleic acids and carbohydrates, but also can combine them with immunostimulators. Liposomes have great potential in the development of new types of vaccines, and much effort has been devoted to enhancing vaccine efficacy in recent years. Different types of immune cells such as macrophages and dendritic cells play an important role in the immune response and in preventing or treating cancer, allergy or many other infectious diseases. Targeting liposome-based delivery systems to certain immune cells and organs is one of the most effective measures in such treatments. Extensive research has shown that liposomes combined with immunostimulators or modified with pattern recognition receptor ligands can target various immune cells and the lymphatic system, thus not only inducing and promoting the desired immune response but also decreasing adverse effects throughout the body and avoiding targeting irrelevant cell types or tissues. Therefore, in this review, we outline some targeting strategies that can be adopted in the design of liposomal vaccines to improve vaccine efficacy, and we summarise the related liposome-based vaccine applications in several diseases. These applications have great potential to treat or prevent some infectious and intractable diseases.

Immunomodulating effect of a seaweed extract from Ulva armoricana in pig: Specific IgG and total IgA in colostrum, milk, and blood.

The transfer of passive immunity from sows to piglets can be improved through the administration of immuno-stimulating products before farrowing. This study evaluated the immuno-stimulating effect of an algal sulfated polysaccharide extract (MSP extract) from the green algae Ulva armoricana when administrated orally to sows at the end of gestation. Four diets were tested: Control (no MSP extract), MSP1 (2 g/day of MSP extract), MSP2 (8 g/day), and MSP3 (16 g/day). The experimental diets were provided in two periods: before the last atrophic rhinitis vaccine booster, and a week before farrowing. Anti-Bordetella IgG antibodies were recorded in blood, colostrum, and milk, and total IgA were measured in colostrum and milk. Titer kinetics between the blood sampled before farrowing and colostrum displayed an increase in specific IgG for MSP3. Moreover, the MSP2 diet increased the level of total IgA in milk compared to the control group. Although the immuno-stimulating effect of MSP extract on piglet performance was not concurrent across the different supplementation levels, the present study supports the use of natural algae extract (MSP) as an immunomodulating solution in swine production.

Phytotherapy as an alternative to conventional antimicrobials: combating microbial resistance.

INTRODUCTION: In the modern antimicrobial era, the rapid spread of resistance to antibiotics and introduction of new and mutating viruses is a global concern. Combating antimicrobial resistant microbes (AMR) requires coordinated international efforts that incorporate new conventional antibiotic development as well as development of alternative drugs with antimicrobial activity, management of existing antimicrobials, and rapid detection of AMR pathogens. Areas covered: This manuscript discusses some conventional strategies to control microbial resistance. The main purpose of the manuscript is to present information on specific herbal medicines that may serve as good treatment alternatives to conventional antimicrobials for infections sensitive to conventional as well as resistant strains of microorganisms. Expert commentary: Identification of potential new antimicrobials is challenging; however, one source for potential structurally diverse and complex antimicrobials are natural products. Natural products may have advantages over other post-germ theory antimicrobials. Many antimicrobial herbal medicines possess simultaneous antibacterial, antifungal, antiprotozoal and/or antiviral properties. Herbal products have the potential to boost host resistance to infections, particularly in immunocompromised patients. Antimicrobial broad-spectrum activity in conjunction with immunostimulatory properties may help to prevent microbial resistance to herbal medicine. As part of the efforts to broaden use of herbal medicines to treat microbial infections, pre-clinical and clinical testing guidelines of these compounds as a whole should be implemented to ensure consistency in formulation, efficacy and safety.

Vaccine adjuvants: Why and how.

Novel vaccine strategies include the so-called subunit vaccines, which encompass only the part of the pathogen to which immune recognition results in protection. The high purity of these vaccines make adverse events less likely, but it also makes the vaccines less immunogenic and therefore potentially less effective. Vaccine adjuvants that increase and modulate the immunogenicity of the vaccine are therefore added to solve this problem. Besides aluminum salts, which have been used in vaccines for 90 years, a number of novel vaccine adjuvants have been included in licensed vaccines over the last 30 years. Increasing insight into immunological mechanisms and how to manipulate them has replaced empirical with rational design of adjuvants, leading to vaccine adjuvants with increased and customized immunogenicity profiles without compromising vaccine safety.

Novel immunostimulators with a thiazolidin-4-one ring promote the immunostimulatory effect of human iNKT cells on the stimulation of Th2-like immune responsiveness via GATA3 activation in vitro.

Invariant natural killer T cells (iNKTs) are important innate immune cells which get involved in various immune responses in both mice and humans. These immune reactions range from self-tolerance to development of autoimmunity and responses to pathogens and tumor development. In this study, we aimed to explore the effects of the novel immunostimulators (CH1b and CH2b) containing thiazolidin-4-one on the functions of human invariant natural killer T cells (iNKTs). First of all, iNKTs in peripheral blood mononuclear cells were expanded with α-Galactosylceramide (α-Galcer) in vitro. Then, the highly purified iNKTs were isolated from PBMCs using magnetic cells sorting (MACS). Next, we investigated the impacts of CH1b and CH2b on proliferation, cytokines production, cytotoxicity, and the associated signaling pathways in iNKT cells. Finally, we found that CH2b could significantly promote the activated iNKTs proliferation, increase the production of Th2 cytokines, and induce Th0 differentiation into Th2 subset via GATA 3 signaling pathway. Besides, CH2b could markedly enhance the cytotoxic ability of the activated iNKTs. Therefore, we concluded that CH2b, a promising candidate immunostimulator, might be used for the treatment of infections, tumors, autoimmune and allergic diseases, and for the correction of Th1/Th2 balance disorders in future.