Cellular mechanisms of graft-versus-host disease in a mouse model

Female CBA/H (H-2k, Mlsb) mice alloimmunized prior to and during syngeneic pregnancy with DBA/2 (H-2d, Mlsa) splenocytes gave rise to offspring which resisted graft-versus-host disease (GVHD) following neonatal intraperitoneal inoculation of high doses of DBA/2 spleen cells. Lymphocytes from GVHD-resistant mice tested after 6 weeks of age were unresponsive to DBA/2 stimulator cells in 72 h mixed lymphocyte cultures. Isotope uptake measured 24 h after culture, however, indicated that a considerable early response was made to DBA/2 which later declined. Proliferative responses to BALB/c were also depressed but no early response to this strain was detected. FACS analysis of T-lymphocyte profiles of the GVHD-resistant CBA/H mice revealed a 100% increase in the Lyt-2+ subpopulation compared to normal CBA/H mice. Significant increases in Lyt-2+ cells were also noted in in vitro cultures of CBA/H lymphocytes responding to GVHD-resistant CBA/H stimulators. Lymphocytes from GVHD-resistant mice suppressed the proliferative responses of normal CBA/H lymphocytes to alloantigenic but not mitogenic stimulation. Suppression of alloantigenic responses were shown to be specific to DBA/2 and did not affect the response to BALB/c stimulator cells, indicating that both anergy and specific suppressor cells were operative in inducing unresponsiveness.