La gD2 coadministrada con el AFCo1 por vía intranasal induce inmunidad protectora contra virus de herpes simple tipo 2 en ratones / gD2 coadministered with AFCo1 by intranasal route induces protective immunity against Virus Herpes Simplex type 2 in mice

La infección por virus herpes simple tipo 2 (VHS-2) continúa siendo un problema de salud mundial. Esta infección es transmitida sexualmente y es la principal causa de úlceras genitales. La prevención de esta enfermedad requiere de la utilización de vacunas mucosales, pues las vacunas parenterales no han sido exitosas. Por otra parte, no existen adyuvantes mucosales, por lo que el desarrollo de estos es esencial para la estrategia de estas vacunas. La administración intranasal (IN) de la glicoproteína D del VHS-2 (gD2), coadministrada con el cocleato (AFCo1+gD2) sería igualmente efectiva con la gD2 incluida (AFCo1-gD2). Se inocularon ratones hembras C57BL/6 por la vía IN con gD2, contenida dentro del cocleato, coadministrada con el cocleato o gD2 sola. Se determinaron los niveles de IgG anti gD2 en suero y lavado vaginal, así como las subclases de IgG anti gD2 por ELISA. Se determinó la respuesta linfoproliferativa en células de bazo, el perfil de citoquinas Th1/Th2, los signos de la enfermedad y la protección frente al reto viral. Se observaron altos títulos de IgG e IgG2c anti gD2 en el suero de los animales inoculados con la gD2 y el AFCo1 como adyuvante. No se observaron diferencias significativas (p>0,05) entre los grupos que recibieron AFCo1+gD2 y los que recibieron AFCo1-gD2. Se observó un perfil de citoquinas tipo Th1 y un 100 por ciento de sobrevida en los grupos que recibieron el AFCo1 como adyuvante de la gD2, mientras que en el grupo que recibió la gD2 sola no se observó protección. Estos resultados indican que la gD2 puede ser utilizada coadministrada con AFCo1 por vía IN como un potencial candidato vacunal contra VHS-2(AU)

Sexually transmitted infections by Herpes Simplex Virus type 2 (HSV-2) are the leading cause of genital ulcers and a major public health problem worldwide. This requires the use of mucosal vaccines, because parenteral vaccines have not been successful. Presently, there are not mucosal adjuvants, for this reason the development of adjuvants is essential for mucosal vaccine strategies. The intranasal (IN) immunizations using HSV-2 glycoprotein D (gD2), coadministered with cochleate (AFCo1+gD2), would be an efficient candidate for future vaccines against HSV2, similar to the gD2 incorporated into AFCo1(AFCo1-gD2). Female C57Bl/6 mice were inoculated with AFCo1-gD2, AFCo1+gD2 or gD2 alone by IN route. The anti gD2 IgG in sera and vaginal fluids and IgG subclasses were measured by ELISA. The lymphoproliferative response in spleen cells, the Th1/Th2 cytokine profile, the protection and the signs of disease against viral challenge were measured. High titers of IgG and IgG2c subclasses were observed in sera of mice that received the gD2 and AFCo1 as adjuvant. No significant differences (p>0.005) were observed in the animals that received AFCo1+gD2 or AFCo1-gD2. a preferential Th1 cytokine profile and 100 percent of survival after challenge were observed in both groups that received the gD2 and AFCo1, while no survival was observed in the group that only received the gD2. These results showed that the gD2 can be used coadministered with AFCo1 by IN route as a potential vaccine candidate against HSV-2(AU)

Influencia de las vías de inmunización mucosales sobre la protección contra herpes simple tipo 2 con el AFCo1 como adyuvante / Influence of mucosal immunization routes on herpes simplex virus type 2 protection with AFCol as adjuvant

Las vacunas mucosales se han planteado como una estrategia prometedora para inducir protección mucosal. El virus herpes simple tipo 2 es uno de los patógenos más frecuentes en el humano transmitidos por vía sexual. Varios candidatos vacunales contra este patógeno se han evaluado, pero no han sido efectivos, por lo que aún no se cuenta con una vacuna profiláctica ni terapéutica. La gD2 es una glicoproteína recombinante y está reportada como uno de los antígenos de importancia vacunal contra este germen. Contamos con el cocleato derivado del proteoliposoma de Neisseria meningitidis serogrupo B (AFCo1) que ha mostrado capacidades adyuvantes por varias vías de inmunización. El objetivo de este trabajo fue evaluar la protección inducida en ratones por el AFCo1-gD2, administrada por diferentes vías mucosales. Se utilizaron ratones hembras C57BL6, los cuales fueron inmunizados por vía intranasal (IN), intravaginal (IVag) o intrarrectal (IR) con AFCo1-gD2 o gD2 sola. Se determinó la IgG anti gD2, la proliferación celular específica, la replicación viral en lavado vaginal, los signos de la enfermedad y la protección frente al reto viral. Se obtuvo respuestas significativas de IgG anti gD2 por todas las vías, aunque la IN mostró los valores más elevados. Se observó proliferación celular en células de animales inmunizados IN e IVag, pero no por vía IR. Se observó la mayor protección (100 por ciento) en los animales inmunizados por vía IN. Se concluye que la vía nasal es la más prometedora en la inducción de protección contra este reto viral(AU)

Mucosal vaccines are a promising strategy to induce mucosal protection. Herpes simplex virus type 2 is the commonest pathogens in the human transmitted by exposure at the genital mucosal surfaces. Many vaccine candidates against this pathogen have been evaluated; but they have not been effective, and neither a prophylactic nor a therapeutic vaccine has been yet obtained. The gD2 is a recombinant glycoprotein and it is reported as one of the antigens of importance for vaccine against this virus. There is the cochleate (AFCo1) derived from proteoliposome of Neisseria meningitidis serogroup B. This cochleate has shown potentialities as adjuvant for several immunization routes. The objective of this study was to evaluate the protection induced in mice by the AFCo1-gD2 administered by different mucosal routes. Female mice C57BL/6 were used and immunized by: intranasal (IN), intravaginal (IVag), or intrarectal (IR) routes with AFCo1-gD2 or gD2 alone. The anti-gD2 IgG and specific cellular proliferation were determined and the viral replication in vaginal wash, the signs of disease and the protection against the viral challenge, were measured too. A significant anti-gD2 IgG response was obtained by all routes, although the IN route showed the highest values. Cellular proliferation was observed in cells of animals immunized IN and IVag route; but not by IR route. In addition, a higher protection (100 percent) in the animals immunized with AFCo1-gD2 by IN route was observed. In conclusion the intranasal is the most promising route in the protection induction against this viral challenge(AU)

Intranasal immunization with a proteoliposome-derived cochleate containing recombinant gD protein confers protective immunity against genital herpes in mice.

The purpose of this study was to investigate the potential of intranasal (IN) immunization with Neisseria meningitides B proteoliposome (AFPL1) and AFPL1-derived cochleate (AFCo1), containing glycoprotein D (gD) of herpes simplex virus type 2 (HSV-2) for induction of protective immunity against genital herpes infection in mice. We could show that IN immunization with both AFPL1 and AFCo1 containing gD induced gD-specific IgG antibody and lymphoproliferative responses. However, IFN-gamma response could only be detected in CD4(+) splenic cells and genital lymph node cells of the AFCo1gD immunized mice upon recall antigen stimulation in vitro. Importantly, IN immunization with AFCo1gD could elicit a complete protection against an otherwise lethal vaginal challenge with HSV-2, while the AFPL1gD immunized mice were only partially protected. Further, we could show that the IFN-gamma response and protective immunity observed after IN immunization with AFCo1gD are mediated via the adaptor molecule myeloid differentiation factor 88. These data may have implications for the development of a mucosal vaccine against genital herpes.