Abstract

Targeting immune responses to the mucosal entry site before pathogen dissemination is critical for protection. We have generated a secreted gp96-fusion protein (gp96-Ig) that mediated strong, antigen specific CD8-CTL expansion in vivo.

The aim of this study was to evaluate the effect of the site of gp96 secretion on mucosal immunity.

Expansion and migration of antigen specific CD8 T cells (adoptively transferred gfp-OT-I responder cells) were measured in response to immunization. Following different routes of administration of gp96-Ig secreting cells (EG7/3T3-OVA) the OT-I response was measured in the peritoneal cavity, spleen, mesenteric lymph nodes and gut mucosa.

OT-I home significantly more efficiently to the gut mucosa if the gp96-Ig was delivered by the intraperitoneal route. Vaginal administration of gp96 also mediated mucosal immunity while subcutaneous and intradermal immunization delivered the weakest response. Secreted gp96-ova complexes recruited OT-I to Peyer’s Patches, intraepithelial compartment and lamina propria within five days. Mucosal OT-I express alpha4beta7, alphaEbeta7/CD103, CCR9, granzyme B, IFNgamma and exhibited effector memory phenotype.

Our results indicate that peritoneally or vaginally cell secreted gp96-Ig induces migration of effector memory cells equipped with cytotoxic molecules into the mucosal compartment by inducing selective expression of gut-homing markers.

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