April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Mixed Chimerism Prolong Mice Keratoplasty Allografts Survival
Author Affiliations & Notes
  • Zhiqiang Pan
    Beijing Tongren Hospital, Beijing Tongren Eye Bank, Beijing, China
  • Yinnan Zhang
    Beijing Tongren Hospital, Beijing Tongren Eye Bank, Beijing, China
  • Ying Jie
    Beijing Tongren Hospital, Beijing Tongren Eye Bank, Beijing, China
  • Footnotes
    Commercial Relationships  Zhiqiang Pan, None; Yinnan Zhang, None; Ying Jie, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 1153. doi:
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      Zhiqiang Pan, Yinnan Zhang, Ying Jie; Mixed Chimerism Prolong Mice Keratoplasty Allografts Survival. Invest. Ophthalmol. Vis. Sci. 2011;52(14):1153.

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

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Abstract

Purpose: : To explore the mixed chimerism and specific immune tolerance induced by superantigen SEB combined with the donor bone marrow transplantation and determine inhibiting mice corneal keratoplasty rejection reaction.

Methods: : The hosts (C57BL/6) were separated into three groups and twenty mice in each group. SEB(75µg/kg, 0.15ml), CTX(80mg/kg.d×3d), and NS(0.15ml)was injected intraperitoneally before the donor (BALB/C) bone marrow transplantation(day 0). Allogenic penetrating keratoplasty models were set up on day 7. The major histocompatibility complex-II (MHC-II) molecules of the donor in periphery blood and spleen were detected by flow-cytometry on day 0 and 14, 28, 56-days after bone marrow transplantation respectively. The host splenocytes were isolated and co-cultured with ConA and donor BALB/C splenocytes separately. The index of cell proliferation of the one way mixed lymphocyte reaction was detected. The percentage of T cell subpopulation (CD4+, CD8+, CD4+CD25+, CD3+NK1.1+) in periphery blood was tested on series timing course. The cornea graft surviving time and the rejection indexes were assessed under splitlamp microscope and photographed. At the 14th day post corneal keratoplasty, 2 eyes from each group were paraffin embedded and sectioned, followed by H&E stain. CD4+T cells and CD8+T cells were immuno-fluorescent labeled, which signal was scanned by confocal microscopy.

Results: : While pretreatment with SEB induced high levels of hematopoietic chimerism on day 14(4.89±0.71%) and(3.19±0.52%)on day 28. While pretreatment with CTX resulted in chimerism(3.10±0.20%), which decreased by day 28 (1.42±0.31%). Pretreatment with NS resulted in only transient chimerism (3.06±0.71%) which almost disappeared by day 28(0.49±0.25). The mean allograft survival time was 18.11±5.35 days in SEB pretreatment group, 11.14±2.41days in CTX pretreatment group, 7.50±1.60 days in NS pretreatment group, which difference has significance(P<0.01). Significant differences in corneal grafts were observed in histological and immunohistochemistry evaluations.

Conclusions: : SEB pretreatment combined with donor bone marrow transplant could efficiently induce the chimerism, which inhibit the allogenic cornea transplant rejection and prolong the cornea graft survival time. It might be related to CD4+ , CD8+ T cell deletion and acquisition of donor-specific tolerance.

Keywords: immune tolerance/privilege • transplantation • cornea: basic science 
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