The effect of Toxoplasma gondii infection on expression of chemokines by rat retinal vascular endothelial cells.
Cells infected by Toxoplasma gondii undergo up-regulation of proinflammatory cytokines, organelle redistribution, and protection from apoptosis. During infection in man, the parasite encysts within the retina, a process that results in retinochoroiditis which can lead to permanent loss of sight. The reasons for the parasite to infect retinal tissue and the mechanisms by which it encysts are not clearly understood. We studied the effect of infection with T. gondii of retinal vascular endothelial cells using the Clontech Atlastrade mark array system in order to elucidate changes in gene expression. We compared hybridization of RNA to the array from infected and uninfected cells at two time points; 2 and 24 h. Exposure to T. gondii after 2 h resulted in change of expression of approximately 6% of genes on the array, including those involved in cell structure, protein and vesicle trafficking, cell-cycle regulation, transcriptional and translational machinery, and apoptosis. Among the genes involved in the inflammatory response, chemokine genes such as GRO1 (Growth Regulated Oncogene 1), MCP-1 (Monocyte Chemotactic Protein-1), FKN (Fractalkine) and RANTES (Regulated upon Activation, Normal T Cell Expressed and Secreted) were found to be up-regulated and protein production was confirmed by ELISA. However after 24 h of infection, GRO1, MCP-1 and FKN were down-regulated, confirmed by RT-PCR. Thus, invasion of retinal vascular endothelium (RVE) cells by T. gondii leads to the production of chemokines important in directing the traffic of inflammatory cells to the infected area.
Knight
B C
BC
Department of Ophthalmology, Rayne Institute, St. Thomas' Hospital London SE1 7EH, United Kingdom. bruce.knight@kcl.ac.uk
Brunton
C L
CL
Modi
N C
NC
Wallace
G R
GR
Stanford
M R
MR
D000818:Animals; D002461:Cell Line, Transformed; D018932:Chemokine CCL2 / Q000096:biosynthesis / Q000235:genetics; D018946:Chemokine CCL5 / Q000096:biosynthesis / Q000235:genetics; D054428:Chemokine CX3CL1; D054360:Chemokine CXCL1; D018925:Chemokines / Q000096:biosynthesis* / Q000235:genetics; D020523:Chemokines, CX3C / Q000096:biosynthesis / Q000235:genetics; D019743:Chemokines, CXC / Q000096:biosynthesis / Q000235:genetics; D004730:Endothelium, Vascular / Q000276:immunology* / Q000378:metabolism / Q000469:parasitology*; D004797:Enzyme-Linked Immunosorbent Assay; D020869:Gene Expression Profiling; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis / Q000235:genetics; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D012171:Retinal Vessels / Q000276:immunology* / Q000378:metabolism / Q000469:parasitology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014122:Toxoplasma / Q000276:immunology*; D015854:Up-Regulation / Q000276:immunology
C491766:Ccl2 protein, rat; D018932:Chemokine CCL2; D018946:Chemokine CCL5; D054428:Chemokine CX3CL1; D054360:Chemokine CXCL1; D018925:Chemokines; D020523:Chemokines, CX3C; D019743:Chemokines, CXC; C515540:Cx3cl1 protein, rat; C513297:Cxcl1 protein, rat; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D012333:RNA, Messenger