Antibody-mediated retinal pericyte injury

Implications for diabetic retinopathy

Yan Li, Dawn Smith, Qing Li, Nader Sheibani, Suber Huang, Timothy Kern, Ram H. Nagaraj, Feng Lin

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

PURPOSE. To test the hypothesis that autoantibodies against retinal pericytes could develop in diabetic retinopathy, and that these autoantibodies could induce retinal pericyte dysfunction/ death via complement. METHODS. Human primary retinal pericytes cultured in media containing normal (5 mM) or high (30 mM) glucose concentrations were incubated with normal human sera in the presence of a retinal pericyte-reactive antibody, then their viability was assessed by a BCECF-based cytotoxicity assay, and their function was assessed by a T-cell proliferation assay. The pericytes were also analyzed by RT-PCR and flow cytometry to detect CD38, an established diabetes-associated cell surface autoantigen. The potential of the anti-CD38 antibodies in inducing pericyte cellular injury was evaluated using the same cytotoxicity assays. In addition, autoantibody-mediated cytotoxicity in mouse retinal pericytes sensitized by sera from mice with developing diabetic retinopathy or control normal mice were also studied. RESULTS. Retinal pericyte-reactive antibodies induced cellular damage by activating complement in the serum. The antibodyinjured pericytes had reduced efficacy in inhibiting T cells. Hyperglycemic culture conditions rendered pericytes more susceptible to antibody-mediated attack. CD38 was expressed in retinal pericytes, and upregulated by TNF-α and IFN-γ, and anti-CD38 antibodies induced pericyte cytotoxicity. Retinal pericytes sensitized with sera from chronic diabetic mice suffered significantly augmented cytotoxicity compared with those sensitized with sera from the control mice. CONCLUSIONS. The autoantibody-initiated complement activation could be a mechanism underlying the loss of function, and eventually, death of retinal pericytes in diabetic patients, suggesting that inhibiting complement activation could be a novel therapeutic approach.

Original languageEnglish (US)
Pages (from-to)5520-5526
Number of pages7
JournalInvestigative Ophthalmology and Visual Science
Volume53
Issue number9
DOIs
StatePublished - Aug 2012
Externally publishedYes

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Pericytes
Diabetic Retinopathy
Antibodies
Wounds and Injuries
Autoantibodies
Serum
Complement Activation
Anti-Idiotypic Antibodies
T-Lymphocytes
Autoantigens

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience
  • Medicine(all)

Cite this

Antibody-mediated retinal pericyte injury : Implications for diabetic retinopathy. / Li, Yan; Smith, Dawn; Li, Qing; Sheibani, Nader; Huang, Suber; Kern, Timothy; Nagaraj, Ram H.; Lin, Feng.

In: Investigative Ophthalmology and Visual Science, Vol. 53, No. 9, 08.2012, p. 5520-5526.

Research output: Contribution to journalArticle

Li, Y, Smith, D, Li, Q, Sheibani, N, Huang, S, Kern, T, Nagaraj, RH & Lin, F 2012, 'Antibody-mediated retinal pericyte injury: Implications for diabetic retinopathy', Investigative Ophthalmology and Visual Science, vol. 53, no. 9, pp. 5520-5526. https://doi.org/10.1167/iovs.12-10010
Li, Yan ; Smith, Dawn ; Li, Qing ; Sheibani, Nader ; Huang, Suber ; Kern, Timothy ; Nagaraj, Ram H. ; Lin, Feng. / Antibody-mediated retinal pericyte injury : Implications for diabetic retinopathy. In: Investigative Ophthalmology and Visual Science. 2012 ; Vol. 53, No. 9. pp. 5520-5526.
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N2 - PURPOSE. To test the hypothesis that autoantibodies against retinal pericytes could develop in diabetic retinopathy, and that these autoantibodies could induce retinal pericyte dysfunction/ death via complement. METHODS. Human primary retinal pericytes cultured in media containing normal (5 mM) or high (30 mM) glucose concentrations were incubated with normal human sera in the presence of a retinal pericyte-reactive antibody, then their viability was assessed by a BCECF-based cytotoxicity assay, and their function was assessed by a T-cell proliferation assay. The pericytes were also analyzed by RT-PCR and flow cytometry to detect CD38, an established diabetes-associated cell surface autoantigen. The potential of the anti-CD38 antibodies in inducing pericyte cellular injury was evaluated using the same cytotoxicity assays. In addition, autoantibody-mediated cytotoxicity in mouse retinal pericytes sensitized by sera from mice with developing diabetic retinopathy or control normal mice were also studied. RESULTS. Retinal pericyte-reactive antibodies induced cellular damage by activating complement in the serum. The antibodyinjured pericytes had reduced efficacy in inhibiting T cells. Hyperglycemic culture conditions rendered pericytes more susceptible to antibody-mediated attack. CD38 was expressed in retinal pericytes, and upregulated by TNF-α and IFN-γ, and anti-CD38 antibodies induced pericyte cytotoxicity. Retinal pericytes sensitized with sera from chronic diabetic mice suffered significantly augmented cytotoxicity compared with those sensitized with sera from the control mice. CONCLUSIONS. The autoantibody-initiated complement activation could be a mechanism underlying the loss of function, and eventually, death of retinal pericytes in diabetic patients, suggesting that inhibiting complement activation could be a novel therapeutic approach.

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