Inmunopotenciadores para la acuicultura / Immune–potentiators for the Aquaculture

La acuicultura es una de las actividades económicas de mayor crecimiento para la producción de alimentos. Unode sus principales retos es la obtención de grandes volúmenes de producción con la mayor calidad posible. Esto conlleva a una reducción de la aplicación de antibióticos y productos quimioterapéuticos. Una de las estrategias más prometedoras es la aplicación de inmunopotenciadores, principalmente en los cultivos intensivos. El objetivode este trabajo fue revisar los principales inmunopotenciadores, así como las tendencias y retos de su uso mundial. Se resumen las particularidades moleculares y funcionales de los mismos y se hace énfasis en los más estudiados: levamisol, ß-glucanos, lipopolisacárido, vitamina C, extractos de plantas y hormonas. Todos estos compuestos de naturaleza heterogénea inciden mayoritariamente en los componentes de la inmunidad innata de los peces, fortaleciendo y potenciando la resistencia a enfermedades; adicionalmente algunos de ellostienen funciones antiestrés y favorecen su crecimiento. Se concluye que los inmunopotenciadores constituyen una estrategia viable para reducir las pérdidas por problemas sanitarios en el sector de la acuicultura; pero aún quedan por solucionar aspectos como la vía de administración y la etapa de inmunización adecuada para cadaespecie y tipo de cultivo(AU)

Aquaculture represents one of the fastest–growing animal food–producing sectors worldwide. One of the mainchallenges is to obtain high–volume production with the highest possible quality, this leads to reduce the use ofantibiotics and chemotherapeutics. A promising solution to these problems is the application of immune–potentiatiors mainly in intensive farming. This article aims to review the main immune–potentiators, as well as thetrends and challenges of global use of them. It summarizes the main molecular and functional characteristics withemphasis on those most studied such as levamisole, ß-glucans, lipopolysaccharide, vitamin C, plant extracts andhormones. All these heterogeneous compounds, mostly affect the innate immunity of fish, strengthening andenhancing disease resistance and some of them additionally have anti–stress effect and promote fish growth. Weconclude that immune–potentiators are a viable strategy to reduce losses for health problems in aquaculture field,however, aspects such as administration route and appropriate immunization phase for each species remains tobe solved(AU)

Pilot scale production of the vaccine adjuvant Proteoliposome derived Cochleates (AFCo1) from Neisseria meningitidis serogroup B.

The use of new adjuvants in vaccine formulations is a subject of current research. Only few parenteral adjuvants have been licensed. We have developed a mucosal and parenteral adjuvant known as AFCo1 (Adjuvant Finlay Cochleate 1, derived from proteoliposomes of N. meningitidis B) using a dialysis procedure to produce them on lab scale. The immunogenicity of the AFCo1 produced by dialysis has been already evaluated, but it was necessary to demonstrate the feasibility of a larger-scale manufacturing process. Therefore, we used a crossflow diafiltration system (CFS) that allows easy scale up to obtain large batches in an aseptic environment. The aim of this work was to produce AFCo1 on pilot scale, while conserving the adjuvant properties. The proteoliposomes (raw material) were resuspended in a buffer containing sodium deoxycholate and were transformed into AFCo1 under the action of a calcium forming buffer. The detergent was removed from the protein solution by diafiltration to a constant volume. In this CFS, we used a hollow fiber cartridge from Amicon (polysulfona cartridge of 10 kDa porosity, 1mm channel diameter of fiber and 0.45 m² area of filtration), allowing production of a batch of up to 20 L. AFCo1 were successfully produced by tangential filtration to pilot scale. The batch passed preliminary stability tests. Nasal immunization of BALB/c mice, induced specific saliva IgA and serum IgG. The induction of Th1 responses were demonstrated by the induction of IgG2a, IFNγ and not IL-5. The adjuvant action over Neisseria (self) antigens and with co-administered (heterologous) antigens such as ovalbumin and a synthetic peptide from haemolytic Streptococcus B was also demonstrated.

Intranasal administration of proteoliposome-derived cochleates from Vibrio cholerae O1 induce mucosal and systemic immune responses in mice.

Conservative estimates place the death toll from cholera at more than 100,000 persons each year. A particulate mucosal vaccine strategy combining antigens and immune stimulator molecules from Vibrio cholerae to overcome this problem is described. Proteoliposomes extracted from V. cholerae O1 were transformed into cochleates (AFCo2, Adjuvant Finlay cochleate 2) through a calcium inducible rotary dialysis method. Light microscopy was carried out and tubules of 16.25+/-4.57 microm in length were observed. Western blots were performed to verify the immunochemical properties of the main AFCo2 incorporated antigens, revealing full recognition of the outer membrane protein U (OmpU), lipopolysaccharide (LPS), and mannose-sensitive hemagglutinin (MSHA) antigens. AFCo2 were administered by the intranasal route using a two or three dose schedule and the immune response against V. cholerae antigens was assessed. Three AFCo2 doses were required to induce significant (p<0.05), antigen specific IgA in saliva (1.34+/-0.135) and feces (0.60+/-0.089). While, two or three doses of AFCo2 or proteoliposomes induce similar specific IgG and vibriocidal activity responses in sera. These results show for the first time that AFCo2 can be obtained from V. cholerae O1 proteoliposomes and have the potential to protect against the pathogen when administered intranasally.

Mucosal immunization using proteoliposome and cochleate structures from Neisseria meningitidis serogroup B induce mucosal and systemic responses.

Most pathogens either invade the body or establish infection in mucosal tissues and represent an enormous challenge for vaccine development by the absence of good mucosal adjuvants. A proteoliposome-derived adjuvant from Neisseria meningitidis serogroup B (AFPL1, Adjuvant Finlay Proteoliposome 1) and its derived cochleate form (Co, AFCo1) contain multiple pathogen-associated molecular patterns as immunopotentiators, and can also serve as delivery systems to elicit a Th1-type immune response. The present studies demonstrate the ability of AFPL1and AFCo1 to induce mucosal and systemic immune responses by different mucosal immunizations routes and significant adjuvant activity for antibody responses of both structures: a microparticle and a nanoparticle with a heterologous antigen. Therefore, we used female mice immunized by intragastric, intravaginal, intranasal or intramuscular routes with both structures alone or incorporated with ovalbumin (OVA). High levels of specific IgG antibody were detected in all sera and in vaginal washes, but specific IgA antibody in external secretions was only detected in mucosally immunized mice. Furthermore, antigen specific IgG1 and IgG2a isotypes were all induced. AFPL1 and AFCo1 are capable of inducing IFN-gamma responses, and chemokine secretions, like MIP-1alpha and MIP-1beta. However, AFCo1 is a better alternative to induce immune responses at mucosal level. Even when we use a heterologous antigen, the AFCo1 response was better than with AFPL1 in inducing mucosal and systemic immune responses. These results support the use of AFCo1 as a potent Th1 inducing adjuvant particularly suitable for mucosal immunization.