CD4+CD25−LAG3+ T cells compose a distinct T cell subset in the human tonsil. Tonsillar CD4+CD25−LAG3+ T cells suppress antibody production more efficiently than CD4+CD25+ T cells. Cell contact is important for suppression by tonsillar CD4+CD25−LAG3+ T cells. Tonsillar CD4+CD25−LAG3+ T cells suppress graft-versus-host disease (GVHD) reactions in humanized mice. Tonsillar CD4+CD25−LAG3+ T cells are a subset of naturally occurring IL-10-producing Tregs.