April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Evidence for Anti-Retinal Auto-Antibodies (AAbs) in the Complement Factor H Chimeric Transgenic (Cfh-Tg) Mouse Model of Age-Related Macular Degeneration (AMD)
Author Affiliations & Notes
  • Alessandro Iannaccone
    Ophthalmology/Hamilton Eye Institute, Univ Tennessee Health Sci Ctr, Memphis, TN
  • Albert H. Alhatem
    Ophthalmology/Hamilton Eye Institute, Univ Tennessee Health Sci Ctr, Memphis, TN
  • Nataliya Lenchik
    Ophthalmology/Hamilton Eye Institute, Univ Tennessee Health Sci Ctr, Memphis, TN
    Medicine/Endocrinology, Univ Tennessee Health Sci Ctr, Memphis, TN
  • Francesco Giorgianni
    Pharmaceutical Sciences, Univ Tennessee Health Sci Ctr, Memphis, TN
  • David D New
    Ophthalmology/Hamilton Eye Institute, Univ Tennessee Health Sci Ctr, Memphis, TN
  • Sarka Beranova-Giorgianni
    Pharmaceutical Sciences, Univ Tennessee Health Sci Ctr, Memphis, TN
  • Ivan Gerling
    Medicine/Endocrinology, Univ Tennessee Health Sci Ctr, Memphis, TN
  • Rafael Ufret-Vincenty
    Ophthalmology, UT Southwestern Medical Center, Dallas, TX
    Neuroscience, UT Southwestern Medical Center, Dallas, TX
  • Marko Radic
    Microbiology, Immunology and Biochemistry, Univ Tennessee Health Sci Ctr, Memphis, TN
  • Footnotes
    Commercial Relationships Alessandro Iannaccone, None; Albert Alhatem, None; Nataliya Lenchik, None; Francesco Giorgianni, None; David New, None; Sarka Beranova-Giorgianni, None; Ivan Gerling, None; Rafael Ufret-Vincenty, None; Marko Radic, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 1205. doi:
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      Alessandro Iannaccone, Albert H. Alhatem, Nataliya Lenchik, Francesco Giorgianni, David D New, Sarka Beranova-Giorgianni, Ivan Gerling, Rafael Ufret-Vincenty, Marko Radic; Evidence for Anti-Retinal Auto-Antibodies (AAbs) in the Complement Factor H Chimeric Transgenic (Cfh-Tg) Mouse Model of Age-Related Macular Degeneration (AMD). Invest. Ophthalmol. Vis. Sci. 2014;55(13):1205.

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

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Abstract

Purpose: To determine if Cfh-Tg mice harboring the H402 Cfh variant (Ufret-Vincenty et al. IOVS 2010; 51: 5878-87) develop circulating AAbs recognizing ocular tissue antigens.

Methods: Methods: We compared sera by Western blots (WBs) from 4 groups: 1) Cfh-Tg expressing the AMD-predisposing H402 Cfh variant (H-Cfh-Tg); 2) H-Cfh-Tg co-expressing the wild-type (WT) transgene of human C-reactive protein (CRP), which interacts with Cfh in inflammation (H-Cfh-Crp-Tg); 3) mice co-expressing the WT Cfh variant (Y402) and the WT Crp gene (Y-Cfh-Crp-Tg); and 4) control adult C57BL/6 mice. Adult C57BL/6 mouse retina/RPE/BM/choroid tissue lysate (10 µg) was loaded on gels, incubated with serum (5µL) and developed. WB band intensity was quantified with the Odyssey system and compared by Kruskall-Wallis test. Immunohistochemistry was done against anti-mouse IgG Ab by fluorescence microscopy following incubation of adult C57BL/6 retina sections with mouse sera. To identify the autoantigens, mouse sera were immunoprecipitated, followed by 2-dimension electrophoresis (2DE) and Mass-Spectrometry (MS) was performed on spots seen on 2DE following previously reported methods (Lenchik et al. ARVO 2013, Abs. 4103).

Results: H-Cfh-Tg mice showed consistently the highest reactivity compared to controls, Y-Cfh-Crp-Tg and H-Cfh-Crp-Tg. Significantly more intense WB bands were seen in H-Cfh-Tg mice at 91kDa (p=0.001), 17kDa and 13kDa (p=0.004), 31kDa (p=0.012), 54kDa (p=0.039), and 22kDa (p=0.045). The outer nuclear (ONL), inner nuclear (INL), and ganglion cell (GCL) layer of adult C57BL/6 mouse retina sections showed strong staining, with an apparent perinuclear pattern, by both H-Cfh and H-Crp-Cfh sera. Differentially reactive spots between H-Cfh-Tg and control mouse sera were observed also on 2DE and preliminary IDs for some of the spots have been obtained via MS, suggesting that antigens involved in apoptotic mechanisms are targeted.

Conclusions: Similar to what we have seen in human AMD sera (Iannaccone et al. Adv. Exp. Med. Biol. 2012; 723:11-6), AAbs recognizing retinal targets develop also in this allele-specific AMD mouse model. This suggests the existence of an autoimmune response to AMD-like retinal degenerative events triggered by the H-Cfh variant, which could be contributing to the pathology observed in the H-Cfh-Tg AMD model as well as in human AMD.

Keywords: 412 age-related macular degeneration • 557 inflammation • 555 immunomodulation/immunoregulation  
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