Preliminary characterization of two thymus-dependent xenoantigens from mouse lymphocytes

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Abstract
Preliminary characterization of 2 mouse thymus-dependent (T) lymphocyte xenoantigens, T25 and T200, which are selectively labeled by lactoperoxidase-catalyzed iodination of T cells, is described. Both molecules are membrane-bound glycoproteins. Fractionation of membrane vesicles prepared from [mouse] BW5147 lymphoma cells by sedimentation through sucrose density gradients show that antigens T25 and T200 are in fractions enriched with plasma membrane. Antigen T200 is partially degraded when viable cells are treated briefly with low concentrations of trypsin. Both molecules are efficiently solubilized in buffers containing sodium deoxycholate or Nonidet P-40, as measured by failure to sediment at 100,000 g for 60 min. Gel filtration on Sepharose 6B showed the presence of aggregated material in Nonidet P-40 extracts which was not found in deoxycholate-solubilized membranes. After solubilization in detergent, antigens T25 and T200 bind to, and may be specifically eluted from columns of pea lectin-Sepharose or concanavalin A-Sepharose. Both molecules are heterogeneous when examined by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulfate. As judged by its binding to columns of pea lectin, at least part of the heterogeneity of mouse thymocyte antigen T25 resides in its carbohydrate moiety.