Mediastinal lymph node CD8alpha- DC initiate antigen presentation following intranasal coadministration of alpha-GalCer.

Our previous study revealed that alpha-galactosylceramide (alpha-Ga1Cer) is a potent nasal vaccine adjuvant inducing both potent humoral and cellular immune responses and affording complete protection against viral infections and tumors. However, the antigen-presenting cells (APC) that are activated by NKT cells and thereby initiate the immune responses following intranasal coadministration of protein antigen and alpha-GalCer are poorly understood. We assessed here where antigen presentation occurs and which APC subset mediates the early stages of immune responses when protein antigen and alpha-GalCer are intranasally administered. We show that dendritic cells (DC), but not B cells, initiated the mucosal immune responses at mediastinal lymph nodes. Of the DC subsets, the CD8 alpha(-)B220(-)CD11c(+) DC subset played the most prominent role in the direct and cross-presentation of protein antigen to naive T cells and in triggering the naive T cells to differentiate into effector T cells. This might be mainly caused by a relatively larger population of CD1d(high) cells of CD8 alpha(-) B220(-)CD11c(+) DC subset than those of other DC subsets. These results indicate that CD8 alpha(-) B220(-)CD11c(+) DC is the principal subset becoming immunogenic after interaction with NKTcells and abrogating tolerance to intranasally administered protein antigen when alpha-GalCer is coadministered as a nasal vaccine adjuvant.