Retinal pigment epithelial cells upregulate expression of complement factors after co-culture with activated T cells

Exp Eye Res. 2011 Mar;92(3):180-8. doi: 10.1016/j.exer.2011.01.003. Epub 2011 Jan 19.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CD28 Antigens / metabolism
  • CD3 Complex / metabolism
  • CD59 Antigens / metabolism
  • Cell Line
  • Clusterin / metabolism
  • Coculture Techniques
  • Complement C3b Inactivator Proteins / metabolism
  • Complement Factor H / metabolism
  • Complement System Proteins / metabolism*
  • Dose-Response Relationship, Drug
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression Regulation / physiology*
  • Humans
  • Immunoblotting
  • Lymphocyte Activation / physiology*
  • Membrane Cofactor Protein / metabolism
  • Oligonucleotide Array Sequence Analysis
  • RNA, Messenger / metabolism
  • Retinal Pigment Epithelium / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • T-Lymphocytes / physiology*
  • Up-Regulation

Substances

  • CD28 Antigens
  • CD3 Complex
  • CD46 protein, human
  • CD59 Antigens
  • CFH protein, human
  • CFHR1 protein, human
  • CLU protein, human
  • Clusterin
  • Complement C3b Inactivator Proteins
  • Membrane Cofactor Protein
  • RNA, Messenger
  • CD59 protein, human
  • Complement Factor H
  • Complement System Proteins