The development of Trypanosoma cruzi in macrophages in vitro. Interaction with lysosomes and host cell fate

SummaryThe interaction between mouse peritoneal macrophages and ‘Y’ strain Trypanosoma cruzi bloodstream forms was studied at optical and electron microscopical levels. The method of marking lysosomes with Thorotrast, either before or after infection of cell monolayers with parasites, revealed that secondary lysosomes fused with phagosomes shortly after trypanosome interiorization. In spite of this, 24 h later most parasites were no longer in a vacuole but lay free within the host cell cytoplasm, multiplying actively. At this time, and up to shortly before 96 h when parasites escaped to the external milieu, most parasitized cells were not lethally injured, as revealed by the Trypan blue dye-exclusion test. Only when parasites were released into the external medium was this situation reversed and infected macrophages took up the dye.

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THE INTERACTION OF SOLUBLE HORSERADISH PEROXIDASE WITH MOUSE PERITONEAL MACROPHAGES IN VITRO

The Journal of Cell Biology ◽

10.1083/jcb.55.1.186 ◽

1972 ◽

Vol 55 (1) ◽

pp. 186-204 ◽

Cited By ~ 251

Author(s):

Ralph M. Steinman ◽

Zanvil A. Cohn

Keyword(s):

Horseradish Peroxidase ◽

Culture Medium ◽

Peritoneal Macrophages ◽

Wide Range ◽

Mouse Macrophages ◽

Secondary Lysosomes ◽

Endocytic Vesicles ◽

In Vitro Interaction ◽

Mouse Peritoneal Macrophages

The in vitro interaction of soluble horseradish peroxidase (HRP) with homogeneous mono layers of mouse macrophages has been studied using sensitive biochemical and cytochemical techniques. The compartmentalization of HRP in extracellular and intracellular sites has been quantitatively evaluated. A significant fraction is bound to a serum-derived layer, which coats the surface of culture vessels and may be removed by appropriate washes. Macrophages interiorize HRP as a solute in pinocytic vesicles without appreciable binding of the glycoprotein to the plasma membrane. Uptake is directly proportional to the concentration of HRP in the culture medium. 1 x 106 cells ingest 0.0025% of the administered load per hr over a wide range of concentrations. Cytochemically, all demonstrable HRP is sequestered within the endocytic vesicles and secondary lysosomes of the vacuolar apparatus. After uptake, the enzymatic activity of HRP is inactivated exponentially with a half-life of 7–9 hr, until enzyme is no longer detectable. When macrophages have pinocytosed trace-labeled HRP-125I, cell-associated isotope disappears with a t ½ of 20–30 hr and they release monoiodotyrosine-125I into the culture medium. We were unable to obtain evidence that significant amounts of HRP (>2%) can be exocytosed after uptake, can exist intact on the cell surface, or can be digested extracellularly. It is difficult to reconcile these observations with several of the postulated mechanisms whereby macrophages are thought to play a prominent role in the induction of an immune response.

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Dipetalonema viteae: ultrastructural study on the in vitro interaction between rat macrophages and microfilariae in the presence of IgE antibody

Parasitology ◽

10.1017/s003118200004186x ◽

1981 ◽

Vol 82 (1) ◽

pp. 55-62 ◽

Cited By ~ 8

Author(s):

M. A. Ouaissi ◽

A. Haque ◽

A. Capron

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Ultrastructural Study ◽

Peritoneal Macrophages ◽

Immune Serum ◽

Sequence Of Events ◽

Transmission Electron ◽

Dipetalonema Viteae ◽

In Vitro Interaction

SUMMARYThe in vitro interaction between rat peritoneal macrophages and Dipetalonema viteae microfilariae in the presence of amicrofilaraemic rat immune serum was studied by transmission electron microscopy. The probable sequence of events leading to the killing of D. viteae microfilaria by macrophages is as follows. (a) Rat peritoneal macrophages in the presence of amicrofilaraemic rat immune serum adhere to the parasite surface, (b) the macrophages extend their pseudopodia around the parasite, (c) the ‘lysosome-like’ granules discharge their contents on to the parasite surface, (d) the lytic activity of these products begins at the parasite surface and (e) subsequent breaking of the microfilarial cuticle occurs, exposing the parasite intracellular material.

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IN VITRO INTERACTION OF MOUSE HEPATITIS VIRUS AND MACROPHAGES FROM GENETICALLY RESISTANT MICE

Journal of Experimental Medicine ◽

10.1084/jem.131.4.843 ◽

1970 ◽

Vol 131 (4) ◽

pp. 843-850 ◽

Cited By ~ 27

Author(s):

Ilan Shif ◽

Frederik B. Bang

Keyword(s):

Virus Replication ◽

Hepatitis Virus ◽

Mouse Hepatitis Virus ◽

Peritoneal Macrophages ◽

Slow Inactivation ◽

Virus Particles ◽

C3h Mice ◽

The Difference ◽

In Vitro Interaction

Peritoneal macrophages from genetically resistant C3H mice and genetically susceptible Princeton (PRI) mice adsorbed the MHV (PRI) strain of mouse hepatitis virus equally well. The difference between the permissive cells and the nonpermissive ones seems to reside in the ability of the former to "eclipse" the virus and, subsequently, support virus replication. C3H cells exposed to low multiplicities of the virus remained intact with no demonstrable viral replication. Virus, taken up by the resistant cells, was protected from heat and underwent slow inactivation while few or no virus particles were released into the medium.

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Inhibition of Host Cell Lysosome Spreading by Trypanosoma cruzi Metacyclic Stage-Specific Surface Molecule gp90 Downregulates Parasite Invasion

Infection and Immunity ◽

10.1128/iai.00302-17 ◽

2017 ◽

Vol 85 (9) ◽

Cited By ~ 8

Author(s):

João Paulo Ferreira Rodrigues ◽

Guilherme Hideki Takahashi Sant'ana ◽

Maria Aparecida Juliano ◽

Nobuko Yoshida

Keyword(s):

Trypanosoma Cruzi ◽

Hela Cell ◽

Specific Surface ◽

Host Cell ◽

Cell Invasion ◽

Target Cell ◽

Content Type ◽

Terminal Domain ◽

Successful Infection

ABSTRACT Successful infection by Trypanosoma cruzi, the agent of Chagas' disease, is critically dependent on host cell invasion by metacyclic trypomastigote (MT) forms. Two main metacyclic stage-specific surface molecules, gp82 and gp90, play determinant roles in target cell invasion in vitro and in oral T. cruzi infection in mice. The structure and properties of gp82, which is highly conserved among T. cruzi strains, are well known. Information on gp90 is still rather sparse. Here, we attempted to fill that gap. gp90, purified from poorly invasive G strain MT and expressing gp90 at high levels, inhibited HeLa cell lysosome spreading and the gp82-mediated internalization of a highly invasive CL strain MT expressing low levels of a diverse gp90 molecule. A recombinant protein containing the conserved C-terminal domain of gp90 exhibited the same properties as the native G strain gp90: it counteracted the host cell lysosome spreading induced by recombinant gp82 and exhibited an inhibitory effect on HeLa cell invasion by CL strain MT. Assays to identify the gp90 sequence associated with the property of downregulating MT invasion, using synthetic peptides spanning the gp90 C-terminal domain, revealed the sequence GVLYTADKEW. These data, plus the findings that lysosome spreading was induced upon HeLa cell interaction with CL strain MT, but not with G strain MT, and that in mixed infection CL strain MT internalization was inhibited by G strain MT, suggest that the inhibition of target cell lysosome spreading is the mechanism by which the gp90 molecule exerts its downregulatory role.

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Improved enzyme-linked immunoadsorbent assay (ELISA) for the study of Trypanosoma cruzi-host cell interaction in vitro

Memórias do Instituto Oswaldo Cruz ◽

10.1590/s0074-02761993000200010 ◽

1993 ◽

Vol 88 (2) ◽

pp. 235-241 ◽

Cited By ~ 2

Author(s):

Mauricio R. M. P. Luz ◽

Maria de Nazaré C. Soeiro ◽

Tania C. Araújo-Jorge

Keyword(s):

Trypanosoma Cruzi ◽

Host Cell ◽

Cell Interaction ◽

Host Cell Interaction

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Infectivity of amastigotes of Trypanosoma cruzi

Revista do Instituto de Medicina Tropical de São Paulo ◽

10.1590/s0036-46651986000400001 ◽

1986 ◽

Vol 28 (4) ◽

pp. 205-212 ◽

Cited By ~ 13

Author(s):

Tecia Ulisses de Carvalho ◽

Wanderley de Souza

Keyword(s):

Trypanosoma Cruzi ◽

Guinea Pig ◽

Cell Line ◽

Peritoneal Macrophages ◽

Mouse Peritoneal Macrophages ◽

Pig Serum

The infectivity amastigotes of Trypanosoma cruzi, isolated from the supernatant of the J774G8 macrophage-like cell line infected with trypomastigotes to normal macrophages in vitro was tested. After a period of 1 h of T. cruzi-macrophage interaction about 2% of the mouse peritoneal macrophages had ingested amastigotes. In contrast 12% of the macrophages had ingested epimastigotes. Treatment of the amastigotes with trypsin did not interfere with their ingestion by macrophages. Once inside the macrophages the amastigotes divided and after some days transformed into trypomastigotes. When i.p. inoculated into mice the amastigotes were highly infective, inducing high levels of parasitaemia and tissue parasitism. As previously described for trypomastigotes, amastigotes were not lysed when incubated in the presence of fresh guinea-pig serum. Contrasting with what has been described for trypomastigotes, the resistance of amastigotes to complement-mediated lysis persisted after treatment with trypsin.

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Interleukin 10 and interferon gamma regulation of experimental Trypanosoma cruzi infection.

Journal of Experimental Medicine ◽

10.1084/jem.175.1.169 ◽

1992 ◽

Vol 175 (1) ◽

pp. 169-174 ◽

Cited By ~ 225

Author(s):

J S Silva ◽

P J Morrissey ◽

K H Grabstein ◽

K M Mohler ◽

D Anderson ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Interferon Gamma ◽

Acute Infection ◽

Interleukin 10 ◽

Peritoneal Macrophages ◽

Biologically Active ◽

Mouse Strains ◽

Ifn Gamma ◽

Cruzi Infection

Studies were undertaken to determine whether interleukin 10, (IL-10) a cytokine shown to inhibit interferon gamma (IFN-gamma) production, was involved in Trypanosoma cruzi infections in mice. Exogenous IFN-gamma protects mice from fatal infection with T. cruzi. Furthermore, resistant B6D2 mice developed fatal T. cruzi infections when treated with neutralizing anti-IFN-gamma monoclonal antibody (mAb). Thus, endogenous as well as exogenous IFN-gamma is important in mediating resistance to this parasite. Because both T. cruzi-susceptible (B6) and -resistant (B6D2) mouse strains produced IFN-gamma during acute infection, we looked for the concomitant production of mediators that could interfere with IFN-gamma-mediated resistance to T. cruzi. We found that IL-10-specific mRNA was produced in the spleens of mice with acute T. cruzi infections. In addition, spleen cell culture supernatants from infected B6 mice, and to a lesser extent B6D2 mice, elaborated an inhibitor(s) of IFN-gamma production. This inhibitor(s) was neutralized by anti-IL-10 mAb. These experiments demonstrated the production of biologically active IL-10 during T. cruzi infection. In further studies in vitro, it was shown that IL-10 blocked the ability of IFN-gamma to inhibit the intracellular replication of T. cruzi in mouse peritoneal macrophages. Thus, in addition to its known ability to inhibit the production of IFN-gamma, IL-10 (cytokine synthesis inhibitory factor), may also inhibit the effects of IFN-gamma. These experiments demonstrate that IL-10 is produced during infection with a protozoan parasite and suggest a regulatory role for this cytokine in the mediation of susceptibility to acute disease.

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Killing in vitro of Trypanosoma cruzi by macrophages from mice immunized with T. cruzi or BCG, and absence of cross-immunity on challege in vivo.

Journal of Experimental Medicine ◽

10.1084/jem.142.2.299 ◽

1975 ◽

Vol 142 (2) ◽

pp. 299-311 ◽

Cited By ~ 71

Author(s):

R Hoff

Keyword(s):

Trypanosoma Cruzi ◽

Peritoneal Macrophages ◽

Cross Immunity

Peritoneal macrophages from T. cruzi-immune mice were resistant to infection in vitro with culture forms of the parasite. Macrophage resistance appeared in infected mice about 21 days postinfection when parasitemia was still rising. Resistance in vitro was nonspecific since macrophages from BCG-immune mice were resistant to T. cruzi, and since macrophages from T. cruzi-immune mice were resistant to infection in vitro with Listeria were not resistant to challenge with T. cruzi even when the parasites were opsonized.

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