Proliferative recruitment of intestinal intraepithelial lymphocytes after microbial colonization of germ-free mice

Intraepithelial lymphocytes (IEL), particularly alpha beta TCR-bearing IEL (alpha beta-IEL), dramatically increase in number after microbial colonization of formerly germ-free (ex-GF) mice (Umesaki et al., Immunology 1993. 79: 32). In this study, the kinetics of expansion of IEL after microbial colonization in ex-GF mice were investigated by the bromodeoxyuridine (BrdUrd) continuous labeling method. In GF mice, gamma delta- and alpha beta-IEL were gradually labeled with BrdUrd, reaching approximately 30% and 15% labeling, respectively, after 10 days of continuous BrdUrd labeling. In conventional (CV) mice, the percentage of BrdUrd-labeled alpha beta-IEL was a little higher than that for gamma delta-IEL. The maximal labeling for alpha beta-IEL and gamma delta-IEL reached 50-60% and 40%, respectively, in 10 days. In the case of conventionalized ex-GF mice, continuous labeling was started 11 days after microbial colonization of GF mice because alpha beta-IEL outnumbered gamma delta-IEL during this period. In this case, 75% of alpha beta-IEL and 67% of gamma delta-IEL were labeled with BrdUrd in 10 days. On the other hand, the apparent half lives of alpha beta-IEL and gamma delta-IEL were 10 and 20 days, respectively in CV mice. These results strongly suggest that the number of IEL, particularly alpha beta-IEL, increases after microbial colonization through recruitment into the cell cycle of a large proportion of IEL or their immediate precursors. The difference in the relative BrdUrd-labeling rate of alpha beta-IEL to that of gamma delta-IEL between germ-free and conventionalized mice indicates a preferential increase in the alpha beta-IEL subset during the course of conventionalization.