Retinal pigment epithelial cells upregulate expression of complement factors after co-culture with activated T cells.
In this study we examined the effect of T cell-derived cytokines on retinal pigment epithelial (RPE) cells with respect to expression of complement components. We used an in vitro co-culture system in which CD3/CD28-activated human T cells were separated from the human RPE cell line (ARPE-19) by a membrane. Differential gene expression in the RPE cells of complement factor genes was identified using gene arrays, and selected gene transcripts were validated by q-RT-PCR. Protein expression was determined by ELISA and immunoblotting. Co-culture with activated T cells increased RPE mRNA and/or protein expression of complement components C3, factors B, H, H-like 1, CD46, CD55, CD59, and clusterin, in a dose-dependent manner. Soluble factors derived from activated T cells are capable of increasing expression of complement components in RPE cells. This is important for the further understanding of inflammatory ocular diseases such as uveitis and age-related macular degeneration.
Eye Research Unit, Department of International Health, Immunology and Microbiology, University of Copenhagen, ISIM 18.3, Blegdamsvej 3, 2100 Copenhagen, Denmark. hjuel@sund.ku.dk
Helene B
HB
Juel
Charlotte
C
Kaestel
Lasse
L
Folkersen
Carsten
C
Faber
Niels H H
NH
Heegaard
Rehannah
R
Borup
Mogens H
MH
Nissen
D018106:CD28 Antigens / Q000378:metabolism; D017252:CD3 Complex / Q000378:metabolism; D018957:CD59 Antigens / Q000378:metabolism; D002460:Cell Line; D051152:Clusterin / Q000378:metabolism; D018920:Coculture Techniques; D003180:Complement C3b Inactivator Proteins / Q000378:metabolism; D017242:Complement Factor H / Q000378:metabolism; D003165:Complement System Proteins / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D015151:Immunoblotting; D008213:Lymphocyte Activation / Q000502:physiology*; D051916:Membrane Cofactor Protein / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000378:metabolism; D055213:Retinal Pigment Epithelium / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000502:physiology*; D015854:Up-Regulation
D018106:CD28 Antigens; D017252:CD3 Complex; C497857:CD46 protein, human; D018957:CD59 Antigens; C493495:CFH protein, human; C517621:CFHR1 protein, human; C495242:CLU protein, human; D051152:Clusterin; D003180:Complement C3b Inactivator Proteins; D051916:Membrane Cofactor Protein; D012333:RNA, Messenger; C051590:CD59 protein, human; D017242:Complement Factor H; D003165:Complement System Proteins