Cultured embryonic retina systems as a model for the study of underlying mechanisms of Toxoplasma gondii infection.
OBJECTIVE
Toxoplasma gondii, the most common cause of retinochoroiditis in humans, is an obligate intracellular protozoan parasite that depends on te host cell's microenvironment to proliferate. Because congenital infection is associated with a higher risk of ocular involvement than a postnatally acquired infection, this study was conducted to investigate the ability of Toxoplasma gondii to infect retinal tissue during development, when cellular environmental changes normally occur.


METHODS
Retinas from 5- to 9-day-old chick embryos were used. Stationary cultures were prepared in 24-well cell culture dishes and maintained at 37 degrees C in DMEM plus 5% fetal bovine serum for 2 to 6 days. Then the wells were infected with 4 x 10(5) tachyzoites. Retina explants and aggregate cell cultures were maintained in DMEM under rotation at 37 degrees C. T. gondii proliferation was measured using [(3)H]-thymidine incorporation after 72 hours. Ornithine and arginine decarboxylase (ODC and ADC) activities were determined by measuring CO(2) production from [1-(14)C]-ornithine and [1-(14)C]-arginine, respectively.


RESULTS
The proliferation of tachyzoites was high in dense, stationary cultures expressing elevated ODC and ADC activity. The addition of ODC or ADC inhibitors reduced T. gondii proliferation by approximately 20% to 40%. As for cultured retina cells, retina explants also allowed T. gondii proliferation whenever ODC activity was high.


CONCLUSIONS
The data suggest a direct correlation between retinal polyamine biosynthesis and the proliferation of T. gondii, in agreement with the observation that individuals infected congenitally have a greater risk of development of toxoplasmic retinochoroiditis.
Moraes
Antonio M M
AM
Instituto de Biofísica Carlos Chagas Filho, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Pessôa
Cristiano N
CN
Vommaro
Rossiane C
RC
De Souza
Wanderley
W
de Mello
Fernando G
FG
Hokoç
Jan Nora
JN
D000818:Animals; D002262:Carboxy-Lyases / Q000378:metabolism; D002478:Cells, Cultured; D002642:Chick Embryo; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007124:Immunoenzyme Techniques; D051379:Mice; D009955:Ornithine Decarboxylase / Q000378:metabolism; D011073:Polyamines / Q000378:metabolism; D012160:Retina / Q000196:embryology* / Q000469:parasitology / Q000648:ultrastructure; D014122:Toxoplasma / Q000502:physiology*; D014124:Toxoplasmosis, Animal / Q000201:enzymology / Q000469:parasitology*; D014125:Toxoplasmosis, Congenital / Q000201:enzymology / Q000469:parasitology*; D014126:Toxoplasmosis, Ocular / Q000201:enzymology / Q000469:parasitology*
D004791:Enzyme Inhibitors; D011073:Polyamines; D002262:Carboxy-Lyases; D009955:Ornithine Decarboxylase; C019722:arginine decarboxylase