Role of the B Cell Antigen Receptor in Antigen Processing and Presentation Involvement of the Transmembrane Region in Intracellular Trafficking of Receptor/Ligand Complexes

To study the role of the B cell Ag receptor (membrane-bound Ig [mIg]) in Ag processing and presentation, we have generated several Ig transfectants that express transfected TEPC-15 idiotype (T15-Id) mIgM with phosphorylcholine (PC)-binding specificity. The wild-type Ig transfectant is able to present a specific Ag (e.g., PC-conjugated hen egg-white lysozyme [PC-HEL]) more efficiently than a nonspecific Ag (HEL) to a T cell hybridoma recognizing an epitope on the HEL molecule. A substitution in the entire spacer, transmembrane, and cytoplasmic region of mIg with an equivalent region of I-Aα chain completely abolishes this mIg-enhanced Ag presentation. Experiments with the wild-type and substituted variant Ig transfectants suggest that this substitution may interfere with normal intracellular trafficking of mIg after cross-linking with specific Ag or antibodies specific for the mIg (anti-T15-Id mAb). Prolonged treatment of the wild-type Ig transfectants with specific Ag or anti-T15-Id mAb reduces the surface expression of T15-Id mIgM and leads to an accumulation of T15-Id mIgM inside the cells. The reduced surface expression and the elevated cytoplasmic accumulation of T15-Id mIgM are not observed in the variant Ig transfectant. Despite the ability of the variant to endocytose ligands similarly to the wild-type Ig transfectant, this variant displays a higher rate of recycling of the mIg/ligand complexes back to the cell surface and a lower rate of degradation of the ligands. These abnormalities may be responsible for the deficiency in mIg-mediated enhancement of Ag presentation in the variant Ig transfectant. Therefore, our results suggest that the transmembrane region of mIg is involved in intracellular trafficking of receptor/ligand complexes and that proper delivery and handling of internalized Ag are required for the enhanced presentation of specific Ag by B cells.