May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Comparison of CD45–positive cells from retina and brain reveals substantial differences in antigen presentation ability
Author Affiliations & Notes
  • D.S. Gregerson
    Department of Ophthalmology, University of Minnesota, Minneapolis, MN
  • T.N. Sam
    Department of Ophthalmology, University of Minnesota, Minneapolis, MN
  • S.W. McPherson
    Department of Ophthalmology, University of Minnesota, Minneapolis, MN
  • Footnotes
    Commercial Relationships  D.S. Gregerson, None; T.N. Sam, None; S.W. McPherson, None.
  • Footnotes
    Support  NIH grant EY11542 and the MN Lions
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 2313. doi:
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      D.S. Gregerson, T.N. Sam, S.W. McPherson; Comparison of CD45–positive cells from retina and brain reveals substantial differences in antigen presentation ability . Invest. Ophthalmol. Vis. Sci. 2004;45(13):2313.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: Although several observations confirm T cell recognition of Ag in retina, there has been no direct demonstration of functional, retinal–derived APC that could provide antigen presentation capable of supporting the intense local responses of experimental autoimmune uveoretinitis. CD45–positive cells from brain have been reported to be efficient APC. To test for antigen presentation by retinal APC, CD45–positive cells were positively selected from retina and brain tested in vitro with naive and antigen–experienced T cells. Methods: CD45–positive cells were isolated from retina and brain, and tested in vitro for functional evidence of Ag presentation to naïve and Ag–experienced CD4 T cells specific for beta–galactosidase (3E9 T cells). Spleen cells were used as positive APC controls. Cultures were harvested for analysis by flow cytometry and cytokine ELISA. Measures of activation included changes in CD4, CD25, CD44, CD45RB, CD62L, CD69, caspase–3 activation, CFSE dilution, size, number of cells recovered, and cytokine production. Results: Ag–experienced T cells responded weakly to Ag presented by retinal CD45–positive cells. Upregulation of activation markers was found, without commensurate increases in cell proliferation or cytokine production. Activation of the retinal cells with IFN–g, anti–CD40 or LPS incrementally increased their activity as APC. Addition of neutralizing antibodies to TGF–b did not reveal suppressed APC activity by the retinal cells. Retinal CD45–positive cells were unable to support Ag–dependent T cell activation and proliferation in naive T cells, unlike splenic APC and CD45–positive cells from brain, which supported potent responses. Instead, addition of retinal CD45–positive cells to co–cultures of naïve 3E9 T cells plus splenic APC inhibited the response of the T cells. The inhibition was not dependent on TCR ligation, but reduced early CD69 expression following Ag–stimulated activation was found. Inhibition was not a result of apoptosis induction, assayed by activation of caspase–3 in the T cells. Conclusions: The APC activity of retinal CD45–positive cells is minimal, and may be directed to limiting the responses of T cells. We speculate that Ag presentation in the retina leading to local inflammation is more likely dependent on recruited APC. Since retina is not associated with tissue equivalents of meninges and choroid plexus, both rich sources of dendritic cells difficult to eliminate from dissected brain, cells from retina appear to better represent the APC activity of fresh, adult CNS parenchymal and perivascular cells.

Keywords: antigen presentation/processing • immunomodulation/immunoregulation • microglia 
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