Xenogeneic graft-versus-host disease (GVHD) models in highly immunodeficient mice are currently being used worldwide to investigate human immune responses against foreign antigens in vivo. However, the individual roles of CD4⁺ and CD8⁺ T cells, and donor/host hematopoietic and nonhematopoietic antigen-presenting cells (APCs) in the induction and development of GVHD have not been fully investigated. In the present study, we comprehensively investigated the immune responses of human T cells and the antigen presentation capacity of donor/host hematopoietic and nonhematopoietic APCs in xenogeneic GVHD models using nonobese diabetic/Shi-scid-IL2rg^null mice. CD4⁺ T cells and, to a lesser extent, CD8⁺ T cells individually mediated potentially lethal GVHD. In addition to inflammatory cytokine production, CD4⁺ T cells also supported the activation and proliferation of CD8⁺ T cells. Using bone marrow chimeras, we demonstrated that host hematopoietic, but not nonhematopoietic, APCs play a critical role in the development of CD4⁺ T cell-mediated GVHD. During early GVHD, we detected 2 distinct populations in memory CD4⁺ T cells. One population was highly activated and proliferated in major histocompatibility complex antigen (MHC)^+/+ mice but not in MHC^−/− mice, indicating alloreactive T cells. The other population showed a less activated and slowly proliferative status regardless of host MHC expression, and was associated with higher susceptibility to apoptosis, indicating nonalloreactive T cells in homeostasis-driven proliferation. These observations are clinically relevant to donor T cell response after allogeneic hematopoietic stem cell transplantation. Our findings provide a better understanding of the immunobiology of humanized mice and support the development of novel options for the prevention and treatment for GVHD.