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Vol. 10, Issue 9, 2891-2904, September 1999

Early Endosomes Are Required for Major Histocompatiblity Complex Class II Transport to Peptide-loading Compartments

Valérie Brachet,^* Gérard Péhau-Arnaudet, Catherine Desaymard, Graça Raposo, and Sebastian Amigorena^*

 ^*Institut National de la Santé et de la Recherche Médicale U520 and  Centre National de la Recherche Scientifique Unite Mixte de Recherche 144, Institut Curie, Section Recherche, 75005 Paris, France

Antigen presentation to CD4⁺ T lymphocytes requires transport of newly synthesized major histocompatibility complex (MHC) class II molecules to the endocytic pathway, where peptide loading occurs. This step is mediated by a signal located in the cytoplasmic tail of the MHC class II-associated Ii chain, which directs the MHC class II-Ii complexes from the trans-Golgi network (TGN) to endosomes. The subcellular machinery responsible for the specific targeting of MHC class II molecules to the endocytic pathway, as well as the first compartments these molecules enter after exit from the TGN, remain unclear. We have designed an original experimental approach to selectively analyze this step of MHC class II transport. Newly synthesized MHC class II molecules were caused to accumulate in the Golgi apparatus and TGN by incubating the cells at 19°C, and early endosomes were functionally inactivated by in vivo cross-linking of transferrin (Tf) receptor-containing endosomes using Tf-HRP complexes and the HRP-insoluble substrate diaminobenzidine. Inactivation of Tf-containing endosomes caused a marked delay in Ii chain degradation, peptide loading, and MHC class II transport to the cell surface. Thus, early endosomes appear to be required for delivery of MHC class II molecules to the endocytic pathway. Under cross-linking conditions, most Ii complexes accumulated in tubules and vesicles devoid of -adaptin and/or mannose-6-phosphate receptor, suggesting an AP1-independent pathway for the delivery of newly synthesized MHC class II molecules from the TGN to endosomes.

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   Corresponding author. E-mail address: sebastian.amigorena{at}curie.fr.

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Molecular Biology of the Cell
Vol. 10, 2891-2904, September 1999
Copyright © 1999 by The American Society for Cell Biology

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