Antigen presentation by Leishmania mexicana-infected macrophages: activation of helper T cells by a model parasite antigen secreted into the parasitophorous vacuole or expressed on the amastigote surface.
Leishmania are protozoan parasites which invade mammalian macrophages and multiply as amastigotes in phagolysosomes (parasitophorous vacuoles). Using L. mexicana and bone marrow-derived macrophages (BMM), the question is addressed whether infected BMM induced to express major histocompatibility complex class II molecules can present defined antigens to specific T helper type 1 cells. As a model antigen, a membrane-bound acid phosphatase (MAP), a minor protein associated with intracellular vesicles in amastigotes, was either overexpressed at the surface of the parasites or overexpressed in a soluble form leading to antigen secretion into the parasitophorous vacuole. Presentation of MAP epitopes by these three types of amastigotes was then compared for macrophages containing live parasites or amastigotes inactivated by drug treatment. It is shown that surface-exposed and secreted MAP can be efficiently presented to T cells by macrophages harboring live amastigotes. Therefore, the parasitophorous vacuole communicates by vesicular membrane traffic with the plasmalemma of the host cell. The intracellular MAP of wild-type cells or the abundant lysosomal cysteine proteinases are not or only inefficiently presented, respectively. After killing of the parasites, abundant proteins such as overexpressed MAP and the cysteine proteinases efficiently stimulate T cells, while wild-type MAP levels are not effective. We conclude that intracellular proteins of intact amastigotes are not available for presentation, while after parasite inactivation, presentation depends on antigen abundance and possibly stability. The cell biological and possible immunological consequences of these results are discussed.
Wolfram
M
M
Max-Planck-Institut für Biologie, Abteilung Membranbiochemie, Tübingen, Germany. mawo@gen.mpib-tuebingen.mpg.de
Fuchs
M
M
Wiese
M
M
Stierhof
Y D
YD
Overath
P
P
D000818:Animals; D017951:Antigen Presentation; D000953:Antigens, Protozoan / Q000096:biosynthesis / Q000276:immunology*; D005260:Female; D007894:Leishmania mexicana / Q000254:growth & development / Q000276:immunology*; D008213:Lymphocyte Activation; D008264:Macrophages / Q000276:immunology* / Q000469:parasitology; D008565:Membrane Proteins / Q000096:biosynthesis*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D013601:T-Lymphocytes / Q000276:immunology* / Q000469:parasitology; D014617:Vacuoles / Q000276:immunology / Q000469:parasitology*
D000953:Antigens, Protozoan; D008565:Membrane Proteins