Journal of Cell Science, Vol 107, Issue 1 69-82, Copyright © 1994 by Company of Biologists

JOURNAL ARTICLES

Distribution of MHC class I and of MHC class II molecules in macrophages infected with Leishmania amazonensis

T Lang, C de Chastellier, C Frehel, R Hellio, P Metezeau, Sd Leao and JC Antoine
Unite d'Immunophysiologie Cellulaire, Institut Pasteur, Paris, France.

Macrophages, being apparently the only cells that in vivo allow the growth of the intracellular pathogen Leishmania, are likely candidates to present antigens to Leishmania-specific CD4+ and CD8+ T lymphocytes, known to be involved in the resolution or in the development of lesions induced by these parasites, and recognizing processed antigens bound to MHC class I and MHC class II molecules, respectively. In the present study, we analysed by confocal microscopy and by immunoelectron microscopy the subcellular distribution of both MHC class I and class II molecules in mouse (Balb/c and C57BL/6 strains) bone marrow-derived macrophages infected for 12 to 48 hours with Leishmania amazonensis amastigotes and activated with gamma interferon to determine the intracellular sites where Leishmania antigens and MHC molecules meet and can possibly interact. Double labelings with anti-MHC molecule antibodies and with either propidium iodide or an anti-amastigote antibody allowed localization of MHC molecules with regard to the endocytic compartments housing Leishmania amastigotes, organelles known as the parasitophorous vacuoles (PV) and which most likely contain the highest concentration of parasite antigens in the host cell. Both uninfected and infected macrophages from Balb/c mice expressed the MHC class I molecules H-2Kd and H-2Dd on their cell surface but no significant amount of these molecules could be detected in the PV, which indicates that, if infected macrophages play a role in the induction of Leishmania-specific CD8+ T lymphocytes, PV are probably not loading compartments for MHC class I molecules. In contrast, MHC class II molecules were found to be associated with the PV membranes as shown previously with microscopic techniques at lower resolution (Antoine et al. Infect. Immun. 59, 764-775, 1991). In addition, we show here that, 48 hours after infection of Balb/c macrophages, in about 90% of PV containing MHC class II molecules, the latter were mainly or solely localized at the attachment zone of amastigotes to PV membranes. This peculiar distribution, especially well demonstrated using confocal microscopy, was confirmed by subcellular fluorescence cytometry of infected macrophages stained for the MHC class II molecules. The following data agree with the idea that PV-associated MHC class II molecules establish specific interactions with plasma membrane components of amastigotes. First, the polarized localization of class II appeared specific to these molecules, since the distribution of the lysosomal glycoproteins Igp110 and Igp120, of the macrosialin (a macrophage-specific marker of endocytic compartments) and of the GTP-binding protein rab7p, shown here as being PV membrane components, was homogeneous.(ABSTRACT TRUNCATED AT 400 WORDS)

This article has been cited by other articles:

				N. Wanasen, C. L. MacLeod, L. G. Ellies, and L. Soong L-Arginine and Cationic Amino Acid Transporter 2B Regulate Growth and Survival of Leishmania amazonensis Amastigotes in Macrophages Infect. Immun., June 1, 2007; 75(6): 2802 - 2810. [Abstract] [Full Text] [PDF]

				M. Olivier, D. J. Gregory, and G. Forget Subversion Mechanisms by Which Leishmania Parasites Can Escape the Host Immune Response: a Signaling Point of View Clin. Microbiol. Rev., April 1, 2005; 18(2): 293 - 305. [Abstract] [Full Text] [PDF]

				E. Prina, S. Z. Abdi, M. Lebastard, E. Perret, N. Winter, and J.-C. Antoine Dendritic cells as host cells for the promastigote and amastigote stages of Leishmania amazonensis: the role of opsonins in parasite uptake and dendritic cell maturation J. Cell Sci., January 15, 2004; 117(2): 315 - 325. [Abstract] [Full Text] [PDF]

				N. Courret, C. Frehel, N. Gouhier, M. Pouchelet, E. Prina, P. Roux, and J.-C. Antoine Biogenesis of Leishmania-harbouring parasitophorous vacuoles following phagocytosis of the metacyclic promastigote or amastigote stages of the parasites J. Cell Sci., January 6, 2002; 115(11): 2303 - 2316. [Abstract] [Full Text] [PDF]

				J. Antoine, T Lang, E Prina, N Courret, and R Hellio H-2M molecules, like MHC class II molecules, are targeted to parasitophorous vacuoles of Leishmania-infected macrophages and internalized by amastigotes of L. amazonensis and L. mexicana J. Cell Sci., January 8, 1999; 112(15): 2559 - 2570. [Abstract] [PDF]

				U. Schaible, P. Schlesinger, T. Steinberg, W. Mangel, T Kobayashi, and D. Russell Parasitophorous vacuoles of Leishmania mexicana acquire macromolecules from the host cell cytosol via two independent routes J. Cell Sci., January 3, 1999; 112(5): 681 - 693. [Abstract] [PDF]

				S De Souza Leao, T Lang, E Prina, R Hellio, and J. Antoine Intracellular Leishmania amazonensis amastigotes internalize and degrade MHC class II molecules of their host cells J. Cell Sci., January 10, 1995; 108(10): 3219 - 3231. [Abstract] [PDF]