Involvement of MAPK activation in chemokine or COX-2 productions by Toxoplasma gondii.
This experiment focused on MAPK activation in host cell invasion and replication of T. gondii, as well as the expression of CC chemokines, MCP-1 and MIP-1 alpha , and enzyme, COX-2/prostaglandin E2 (PGE2) in infected cells via western blot, [3H]-uracil incorporation assay, ELISA and RT-PCR. The phosphorylation of ERK1/2 and p38 in infected HeLa cells was detected at 1 hr and/or 6 hr postinfection (PI). Tachyzoite proliferation was reduced by p38 or JNK MAPK inhibitors. MCP-1 secretion was enhanced in infected peritoneal macrophages at 6 hr PI. MIP-1 alpha mRNA was increased in macrophages at 18 hr PI. MCP-1 and MIP-1 alpha were reduced after treatment with inhibitors of ERK1/2 and JNK MAPKs. COX-2 mRNA gradually increased in infected RAW 264.7 cells and the secretion of COX-2 peaked at 6 hr PI. The inhibitor of JNK suppressed COX-2 expression. PGE2 from infected RAW 264.7 cells was increased and synthesis was suppressed by PD98059, SB203580, and SP600125. In this study, the activation of p38, JNK and/or ERK1/2 MAPKs occurred during the invasion and proliferation of T. gondii tachyzoites in HeLa cells. Also, increased secretion and expression of MCP-1, MIP-1 alpha , COX-2 and PGE2 were detected in infected macrophages, and appeared to occur via MAPK signaling pathways.
Kim
Ji-Young
JY
Department of Parasitology, Institute of Biomedical Science, Hanyang University College of Medicine, Seoul, Korea.
Ahn
Myoung-Hee
MH
Song
Hyun-Ouk
HO
Choi
Jong-Hak
JH
Ryu
Jae-Sook
JS
Min
Duk-Young
DY
Cho
Myung-Hwan
MH
D000818:Animals; D018925:Chemokines / Q000096:biosynthesis*; D051546:Cyclooxygenase 2 / Q000096:biosynthesis*; D004789:Enzyme Activation; D006367:HeLa Cells; D006801:Humans; D017737:Macrophages, Peritoneal / Q000201:enzymology / Q000276:immunology / Q000469:parasitology; D051379:Mice; D008807:Mice, Inbred BALB C; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D014122:Toxoplasma / Q000276:immunology* / Q000378:metabolism*; D014123:Toxoplasmosis / Q000201:enzymology* / Q000276:immunology*
D018925:Chemokines; D051546:Cyclooxygenase 2; D020928:Mitogen-Activated Protein Kinases