Expression analysis of immune response genes of Müller cells infected with Toxoplasma gondii.
OBJECTIVE
To investigate changes in immune response genes following Toxoplasma gondii infection of Müller cells.


METHODS
Human Müller cells were infected or mock infected with two strains of T. gondii (RH and Prugniaud). RNA and supernatants were collected from infected and uninfected cells at 2 and 24 h. RNA from the two time points were compared using a custom made DNA microarray. Real time PCR or human cytokine antibody array was used to confirm up-regulation of immune molecules.


RESULTS
Gene expression in infected cells showed up-regulation of CCL2, IL-6, CXCL8, and CXCL2. CCL2 and CXCL2 gene expression was confirmed by real time PCR. IL-6 and CXCL8 protein production was confirmed by a cytokine antibody array. IL-4 production was observed by cytokine antibody array but not by DNA microarray. In contrast, infection with T. gondii did not induce interferon-gamma (IFNgamma) and IL-12 expression, molecules conventionally associated with the inter-conversion of tachyzoite to bradyzoite.


CONCLUSIONS
These results indicate that while in vitro infected Müller cells may be capable of inducing an immune response by attracting blood-borne leucocytes, they do not appear able to directly control the proliferation of T. gondii.
Knight
B C
BC
Department of Ophthalmology, Rayne Institute, St Thomas' Hospital, London, UK.
Kissane
S
S
Falciani
F
F
Salmon
M
M
Stanford
M R
MR
Wallace
G R
GR
D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D015870:Gene Expression / Q000276:immunology*; D020869:Gene Expression Profiling; D005802:Genes, MHC Class II / Q000502:physiology*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014122:Toxoplasma / Q000276:immunology; D014123:Toxoplasmosis / Q000276:immunology*
D016207:Cytokines