Invasion of Toxoplasma gondii occurs by active penetration of the host cell.

Toxoplasma gondii is an obligate intracellular parasite that infects a wide variety of vertebrate cells including macrophages. We have used a combination of video microscopy and fluorescence localization to examine the entry of Toxoplasma into macrophages and nonphagocytic host cells. Toxoplasma actively invaded host cells without inducing host cell membrane ruffling, actin microfilament reorganization, or tyrosine phosphorylation of host proteins. Invasion occurred rapidly and within 25-40 seconds the parasite penetrated into a tight-fitting vacuole formed by invagination of the plasma membrane. In contrast, during phagocytosis of Toxoplasma, extensive membrane ruffling captured the parasite in a loose-fitting phagosome that formed over a period of 2-4 minutes. Phagocytosis involved both reorganization of the host cytoskeleton and tyrosine phosphorylation of host proteins. In some cases, parasites that were first internalized by phagocytosis, were able to escape from the phagosome by a process analogous to invasion. These studies reveal that active penetration of the host cell by Toxoplasma is fundamentally different from phagocytosis or induced endocytic uptake. The novel ability to penetrate the host cell likely contributes to the capability of Toxoplasma-containing vacuoles to avoid endocytic processing.

[1]  Stephen J. Smith,et al.  Ruffles induced by Salmonella and other stimuli direct macropinocytosis of bacteria , 1993, Nature.

[2]  J. Remington,et al.  Effect of cytochalasin D on Toxoplasma gondii cell entry , 1978, Infection and immunity.

[3]  B. Finlay,et al.  Salmonella typhimurium invasion of epithelial cells: role of induced host cell tyrosine protein phosphorylation , 1994, Infection and immunity.

[4]  S. Greenberg,et al.  Tyrosine phosphorylation is required for Fc receptor-mediated phagocytosis in mouse macrophages , 1993, The Journal of experimental medicine.

[5]  S. Greenberg,et al.  Colocalization of F-actin and talin during Fc receptor-mediated phagocytosis in mouse macrophages , 1990, The Journal of experimental medicine.

[6]  E. Porchet-Henneré,et al.  Relations entre toxoplasma et sa cellule-hôte , 1983 .

[7]  S. Greenberg,et al.  Tyrosine phosphorylation of the gamma subunit of Fc gamma receptors, p72syk, and paxillin during Fc receptor-mediated phagocytosis in macrophages. , 1994, The Journal of biological chemistry.

[8]  S. Miller,et al.  Salmonella stimulate macrophage macropinocytosis and persist within spacious phagosomes , 1994, The Journal of experimental medicine.

[9]  L. Miller,et al.  Freeze-fracture study on the erythrocyte membrane during malarial parasite invasion , 1981, The Journal of cell biology.

[10]  R. Werk How does Toxoplasma gondii enter host cells? , 1985, Reviews of infectious diseases.

[11]  S. Silverstein,et al.  Studies on the mechanism of phagocytosis. I. Requirements for circumferential attachment of particle-bound ligands to specific receptors on the macrophage plasma membrane , 1975, The Journal of experimental medicine.

[12]  L. Sibley,et al.  Phagosome acidification blocked by intracellular Toxoplasma gondii , 1985, Nature.

[13]  L. Sibley,et al.  Lymphokine activation of J774G8 cells and mouse peritoneal macrophages challenged with Toxoplasma gondii , 1985, Infection and immunity.

[14]  J. Schwartzman,et al.  Immunofluorescent localization of myosin at the anterior pole of the coccidian, Toxoplasma gondii. , 1983, The Journal of protozoology.

[15]  W. de Souza,et al.  Cytochemical localization of plasma membrane enzyme markers during interiorization of tachyzoites of Toxoplasma gondii by macrophages. , 1989, The Journal of protozoology.

[16]  K. Joiner,et al.  Toxoplasma gondii: fusion competence of parasitophorous vacuoles in Fc receptor-transfected fibroblasts. , 1990, Science.

[17]  J. Swanson,et al.  Macropinosome maturation and fusion with tubular lysosomes in macrophages , 1993, The Journal of cell biology.

[18]  T. Asai,et al.  Transmission and scanning electron microscopy of host cell entry by Toxoplasma gondii. , 1977, The American journal of pathology.

[19]  L. Miller,et al.  Erythrocyte entry by malarial parasites. A moving junction between erythrocyte and parasite , 1978, The Journal of cell biology.

[20]  J. Dvorak,et al.  The origin of parasitophorous vacuole membrane lipids in malaria-infected erythrocytes. , 1993, Journal of cell science.

[21]  B. Finlay,et al.  Cytoskeletal rearrangements accompanying salmonella entry into epithelial cells. , 1991, Journal of cell science.

[22]  W. Bommer The life cycle of virulent toxoplasma in cell cultures. , 1969, Australian Journal of Experimental Biology and Medical Science.

[23]  J. G. Hirsch,et al.  THE ABSENCE OF LYSOSOMAL FUSION WITH PHAGOCYTIC VACUOLES CONTAINING LIVING PARASITES* , 2003 .

[24]  G. Bancroft,et al.  Cytokine enhancement of complement‐dependent phagocytosis by macrophages: synergy of tumor necrosis factor‐α and granulocyte‐macrophage colony‐stimulating factor for phagocytosis of Cryptococcus neoformans , 1992, European journal of immunology.

[25]  V. Tsai,et al.  Failure to trigger the oxidative metabolic burst by normal macrophages: possible mechanism for survival of intracellular pathogens , 1980, The Journal of experimental medicine.

[26]  S. Falkow,et al.  Signal transduction in the mammalian cell during bacterial attachment and entry , 1993, Cell.

[27]  J. G. Hirsch,et al.  THE INTERACTION BETWEEN TOXOPLASMA GONDII AND MAMMALIAN CELLS , 1972, The Journal of experimental medicine.

[28]  G. O'Connor,et al.  Penetration of mouse peritoneal macrophages by the protozoon Toxoplasma gondii. New evidence for active invasion and phagocytosis. , 1981, Laboratory investigation; a journal of technical methods and pathology.

[29]  S Falkow,et al.  The interaction of bacteria with mammalian cells. , 1992, Annual review of cell biology.

[30]  Nichols Ba,et al.  Penetration of mouse peritoneal macrophages by the protozoon Toxoplasma gondii. New evidence for active invasion and phagocytosis. , 1981 .