evaluation of ovalbumin-specific antibodies in mice after intramuscular administration of cationic nanoemulsion&#45;encapsulated retinoic acid

Evaluating Immunogenicity of Nanoemulsion&amp;#45;Encapsulated Retinoic Acid in Mice

Adjuvants are often used to enhance the efficacy of a vaccine by stimulating the immune system to respond more strongly to the vaccine, thereby increasing immunity to a particular pathogen1. Nanoemulsion (NE) adjuvant refers to a colloidal dispersion system with thermodynamic stability by emulsifying a certain proportion of oil phase and aqueous phase to produce an emulsion in the form of water-in-oil (W/O) or oil-in-water (O/W)2. O/W nanoemulsion adjuvant can produce cytokines and chemokines at the injection site, induce the rapid aggregation and proliferation of important immune cells such as monocytes, neutrophils, and eosinophils, and enhance the immune response, and improve the immunogenicity of antigens3. Currently, three nanoemulsion adjuvants (MF59, AS03, and AF03) have been licensed for use in vaccines and have shown good safety and efficacy4.
However, mucosal immunity has been poorly addressed by the currently licensed adjuvant formulations in conventional parenteral vaccination5. Retinoic acid (RA) has been reported to induce intestinal homing of immune cells, but its low polarity, poor solubility in water, and poor light and thermal stability limit its use for robust enteric vaccination. Nanoemulsions offer opportunities to increase the bioavailability of highly lipophilic drugs, and the oil core of O/W emulsion adjuvants is suitable for dissolving non-polar substances such as RA6. Therefore, nanoemulsions can be used as carriers for RA in order to achieve the dual response effect of systemic immunity and mucosal immunity.
Compared to neutral or anionic delivery systems, cationic delivery systems have relatively efficient antigen encapsulation and delivery capabilities, which can enhance the immunogenicity of antigens7,8,9. The cationic surface charge of a variety of adjuvant systems is important for their adjuvant effects10,11,12. The cationic charge is an important factor in prolonging vaccine retention at the injection site, increasing antigen presentation and prolonging the stimulation of cellular immunity in the body12.
Based on the above considerations, we developed a cationic nanoemulsion to effectively co-deliver RA and antigens. The particle size and zeta potential of the nanoemulsion were determined using dynamic light scattering (DLS), and the systemic and mucosal immune responses of the nanoemulsion combined with OVA were evaluated by intramuscular injection13.

Author Spotlight: Development and Evaluation of a Cationic Nanoemulsion&amp;#45;Encapsulated Retinoic Acid System for Mucosal Vaccination

Cationic Nanoemulsion-Encapsulated Retinoic Acid as an Adjuvant to Promote OVA&#45;Specific Systemic and Mucosal Responses

We are working on a mucosal vaccine that produces an immune response as the set of infection, which has the potential to prevent infection and interrupt transmission. We have developed a catatonic nanoemulsion-encapsulated retinoic acid delivery system immunization with ovalbumin by intramuscular injection, which triggers both systemic and mucosal immune responses. In future studies, animal models of intestinal-related diseases can be used to further evaluate the effect and the mechanism of a cationic nanoemulsion-encapsulated retinoic acid in enhancing vaccine protection.

Evaluation of Ovalbumin-Specific Antibodies in Mice After Intramuscular Administration of Cationic Nanoemulsion&#45;Encapsulated Retinoic Acid

evaluation-ovalbumin-specific-antibodies-mice-after-intramuscular

Research

JoVE Journal

Immunology and Infection

66 Views.  Third Military Medical University. Per iniziare il rivestimento dell'antigene, diluire l'ovoalbumina a cinque microgrammi per millilitro con un tampone di rivestimento. Rivestire una piastra ELISA a 96 pozzetti con 100 microlitri di soluzione di ovoalbumina per pozzetto durante la notte a quattro gradi Celsius. Il giorno successivo, lavare i pozzetti cinque volte con PBST.Bloccare i pozzetti con 250 microlitri di PBST all'1% di BSA a 37 gradi Celsius per due ore. Per preparare una diluizione di siero 1000 X, diluire pri...

Video: Valutazione degli anticorpi specifici dell'ovoalbumina nei topi dopo somministrazione intramuscolare di acido retinoico incapsulato con nanoemulsione cationica

Cationic nanostructures have emerged as an adjuvant and antigen delivery system that enhances dendritic cell maturation, ROS generation, and antigen uptake and then promotes antigen-specific immune responses. In recent years, retinoic acid (RA) has received increasing attention due to its effect in activating the mucosal immune response; however, in order to use RA as a mucosal adjuvant, it is necessary to solve the problem of its dissolution, loading, and delivery. Here, we describe a cationic nanoemulsion-encapsulated retinoic acid (CNE-RA) delivery system composed of the cationic lipid 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOTAP), retinoic acid, squalene as the oil phase, polysorbate 80 as surfactant, and sorbitan trioleate 85 as co-surfactant. Its physical and chemical properties were characterized using dynamic light scattering and a spectrophotometer. Immunization of mice with the mixture of antigen (ovalbumin, OVA) and CNE-RA significantly elevated the levels of anti-OVA secretory immunoglobulin A (sIgA) in vaginal lavage fluid and the small intestinal lavage fluid of mice compared with OVA alone. This protocol describes a detailed method for the preparation, characterization, and evaluation of the adjuvant effect of CNE-RA.

In this protocol, we developed a cationic nanoemulsion-encapsulated retinoic acid (RA) to be used as an adjuvant to promote antigen-specific systemic and mucosal responses. By adding the FDA-approved RA to the nanoemulsion, antigen-specific sIgA was promoted in the vagina and small intestine after intramuscular injection of the nanoemulsion.

Cationic nanostructures have emerged as an adjuvant and antigen delivery system that enhances dendritic cell maturation, ROS generation, and antigen ...