Endocytosis, intracellular trafficking, and processing of membrane IgG and monovalent antigen/membrane IgG complexes in B lymphocytes

Human B lymphoblastoid cell lines specific for tetanus toxin/toxoid were used in our earlier studies to demonstrate the rapid endocytosis of monovalent Ag and its processing, as a complex with mIgG. Here we show that the mIgGR for Ag is endocytosed in the presence or absence of Ag and that at any given time about 60% of this recycling pool of membrane (m) Ig is inside the cell. During the earliest detectable stages of Ag processing a high proportion of Ag fragments resolved on SDS gels were bound to intact mIg. However at later times, as fragmented Ag accumulated, the fragments were precipitable only with antibodies against the Fab region of mIgG indicating proteolytic fragmentation of this receptor. Fractionation of cell homogenates on self-forming Percoll gradients revealed that at least two compartments are involved in Ag processing: a low density endosome compartment and a dense 'late endosome'/lysosomal compartment. The spectrum of Ag fragments observed in each fraction differed: fragments produced at later times during processing were detected only in the late endosome/lysosomal fraction whereas the earliest observed fragments were found both in this fraction and in the low density fractions. Monovalent Ag/mIgG complexes appear to have an increased probability, compared to unoccupied mIgG, of targeting to proteolytically active compartments leading to processing of the Ag/mIg complex and to accelerated degradation of the mIgG.