Role of annexins in endocytosis of antigens in immature human dendritic cells.
We have evaluated the uptake of a soluble protein antigen, denitrophenylated human serum albumin (DNP-HSA), and two different intracellular bacteria; Chlamydia trachomatis serovar L2 and Mycobacterium tuberculosis strain H37Ra, by immature human dendritic cells. These were generated by culturing progenitor cells from blood in the presence of cytokines (granulocyte-macrophage colony-stimulating factor and interleukin-4). Dendritic cells play a crucial part in antigen presentation for the induction of T-cell-dependent immune responses in various tissues. Recently, macropinocytic and phagocytic activity has been shown for immature dendritic cells of mouse, rat and human origin. In the present study, macropinocytosis characterized the uptake of the soluble protein-antigen DNP-HSA, whereas the C. trachomatis were ingested via receptor-mediated endocytosis in coated pits, and opsonized M. tuberculosis via phagocytosis. To follow the intracellular routes of the antigens, their positions were compared with the localization of annexins, a family of Ca(2+)-and phospholipid-binding proteins, involved in membrane fusion, aggregation and transport of different vesicles. To elucidate further the intracellular pathway of the antigens, two other proteins, lysosome-associated membrane protein-1 (LAMP-1) and cathepsin D, were labelled. They are known to colocalize with major histocompatibility complex class II compartments in the immature dendritic cells. We observed a distinct translocation of annexin V to DNP-HSA containing endosomes, and annexin III to vesicles with C. trachomatis. Furthermore, annexin III, IV and V redistributed to phagosomes with M. tuberculosis. Both LAMP-1 and cathepsin D colocalized with DNP-HSA endosomes, and with phagosomes with M. tuberculosis. Thus, immature human dendritic cells have the capacity to phagocytose. Moreover, the handling of these antigens by dendritic cells may represent three distinct intracellular pathways, albeit some properties and compartments are shared.
Larsson
M
M
Department of Health and Environment, Faculty of Health Sciences, Linköping University, Sweden.
Majeed
M
M
Ernst
J D
JD
Magnusson
K E
KE
Stendahl
O
O
Forsum
U
U
D017302:Annexins / Q000276:immunology* / Q000378:metabolism; D015703:Antigens, CD / Q000378:metabolism; D002402:Cathepsin D / Q000378:metabolism; D018929:Cell Culture Techniques; D002690:Chlamydia Infections / Q000276:immunology; D002692:Chlamydia trachomatis; D003713:Dendritic Cells / Q000276:immunology*; D004140:Dinitrophenols / Q000276:immunology; D004705:Endocytosis / Q000276:immunology*; D006801:Humans; D016130:Immunophenotyping; D051907:Lysosomal Membrane Proteins; D008562:Membrane Glycoproteins / Q000378:metabolism; D018613:Microscopy, Confocal; D012709:Serum Albumin / Q000276:immunology; D014376:Tuberculosis / Q000276:immunology
D017302:Annexins; D015703:Antigens, CD; D004140:Dinitrophenols; D051907:Lysosomal Membrane Proteins; D008562:Membrane Glycoproteins; D012709:Serum Albumin; C058064:dinitrophenyl-human serum albumin conjugate; D002402:Cathepsin D