Innate immune cells, known as invariant natural killer T (iNKT) cells are recognized for their ability to secrete cytokines and immune mediators. These cells identify glycolipids, whether of self or microbial origin, when presented by CD1d. During intestinal inflammation, iNKT cells can lead to protection or contribute to pathogenesis. While colonic iNKT are believed to be fixed and unalterable later in life, our study challenges this notion. Our previous research showed that Goblet cells (GCs) or/and goblet cell-associated passages (GAPs) transport luminal antigens and maintain peripheral T regulatory cells during the steady state in the small intestine (SI). However, GAPs are absent in the colon due to a high microbial load. We found that inducing GAPs in the colon of adult mice led to a substantial expansion of iNKT cells. Deletion of CD1d on GCs hindered iNKT cell expansion, highlighting the crucial role of colonic GCs in presenting glycolipids to iNKT cells. Single-cell RNA sequencing of colonic-iNKT cells revealed a notable increase in the iNKT2 and iNKT1 subsets following colonic GAP induction. Furthermore, the expanded iNKT cells exhibited persistence and conferred protection in the context of DSS-induced colitis. Together, these findings suggest that adult colonic iNKT cells have the capacity to expand in a GAP-dependent and GC-CD1d-dependent manner, conferring protection in specific colitis models.