Mechanisms involved in antithymocyte globulin immunosuppressive activity in a nonhuman primate model.

BACKGROUND: The mechanisms of action of polyclonal antithymocyte globulins (ATGs) are still poorly understood and the selection of doses used in different clinical applications (prevention or treatment of acute rejection in organ allografts, treatment of graft-versus-host disease, or conditioning for allogeneic stem cell transplantation) remains empirical. Low T-cell counts are usually achieved in peripheral blood during ATG treatment but the extent of T-cell depletion in lymphoid tissues is unknown. METHODS: Experiments were conducted in cynomolgus monkeys using Thymoglobuline at low (1 mg/kg), high (5 mg/kg), and very high (20 mg/kg) doses. RESULTS: ATG treatment induced a dose-dependent lymphocytopenia in the blood and a dose-dependent T-cell depletion in spleen and lymph nodes but not in the thymus, indicating a limited access of ATG to this organ. T-cell apoptosis in peripheral lymphoid tissues was the main mechanism of depletion. Remaining T cells in peripheral lymphoid organs were coated by antibodies and had down-modulated surface expression of CD2, CD3, CD4, and CD8 molecules, whereas their responsiveness in mixed leukocyte reaction was impaired. The survival of MHC-mismatched skin and heart allografts was prolonged in a dose-dependent fashion, despite the occurrence of a strong anti-ATG antibody response resulting in the rapid clearance of circulating ATGs. CONCLUSION: The results indicate that T-cell depletion is achieved rapidly and primarily in peripheral lymphoid tissues at high ATG dosage. Short ATG treatments could therefore be clinically evaluated when major peripheral T-cell depletion is required.

Human natural antibodies to porcine platelets.

The specificity of human natural antibodies directed against blood cells from pigs was investigated by ELISA and immunoblotting. Both IgG and IgM were identified as xenoantibodies reacting with pig platelets adsorbed to microplates. The antibodies could be absorbed on platelets as well as on RBC, suggesting that the corresponding antigens are expressed on the surface of a variety of cells. Galactose (20 mM) and melibiose (10 mM) partially inhibited (approximately 50%) the binding of antibodies to platelets, whereas lactose and cellobiose (300 mM) did not. On immunoblots, platelet glycoproteins of 115, 125, 135, 180, and 210 kDa were specifically revealed with human sera diluted 1/20. In contrast with the results obtained by ELISA, xenoantibodies reactive with blotted glycoproteins were of the IgM class and the binding was not significantly inhibited by galactose or melibiose. "Anti-Gal" antibodies, purified from human sera by affinity chromatography on a melibiose-Sepharose immunoabsorbent, represented only a minor portion of the antibodies reactive with porcine platelets. Purified anti-Gal antibodies bound to the 115- and 135-kDa components, whereas the antibodies in the nonretained fraction revealed the 125-kDa molecule. As deduced from these data, human serum contains natural antibodies of both IgG and IgM classes directed to several porcine antigens. Gal-reactive structures were identified on the 115- and 135-kDa platelet glycoproteins, which might be homologous to their counterpart on endothelial cells. Also, the present work suggests that a majority of the natural antibodies reacted with other unidentified structures.