Immunochemical evidence for multiple beta units of the class II molecule in the rat

The RT1-B/D region-associated antigens which were serologically defined in the previous study (Ohhashi et al., 1981), were partially purified from membranes of a rat B cell leukaemia, KNL-14. Sequential immunoprecipitation test, with the partially purified 125I-B/Dak preparation using four different rat alloantisera, including a monoclonal antibody, disclosed three distinctive populations of beta units of the class II molecules. Highly purified beta units of three discriminable class II molecules were shown to have different structural properties in terms of molecular weights and of electrophoretic profiles on the isoelectric focusing. The beta units shifted to a position of higher molecular weight on SDS-PAGE under reducing condition, thus suggesting to carry intradisulfide bonds. Furthermore, the highly purified beta units cross-reacted with murine anti-Ia sera. The rebinding test revealed that at least two discriminable species of beta units cross-react with anti-I-Ak monoclonal antibody, whereas beta units purified by binding with the 1E4 monoclonal antibody cross-reacted with anti-I-Ab and/or anti-I-Ad antiserum. On the basis of structural and antigenic properties, we have postulated that the rat class II region can be divided into at least three subregions, each containing a locus which encodes a distinctive beta unit of the class II molecule.