IRBP-specific Th1 cells from peripheral blood were predominant in the experimental autoimmune uveitis

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Abstract

Experimental autoimmune uveitis (EAU) is a Th1-cell-mediated autoimmune disease. In this study, the correlation between IRBP-specific Th1 cells in PBLs and the histological grading in the eyes was evaluated kinetically during EAU induction. EAU was induced in B10.A mice with IRBP immunization and the eyes were enucleated for histological examination on days 0, 3, 7, 15, and 21 after immunization. To determine the Th1-cell-mediated immune response, Th1 cytokines (IL-12p40 and IFN-γ) were measured by RT-PCR in inflamed eyes. At mean time, CD4+ and IFN-γ+ double positive T cells (Th1 cells) from PBLs were analyzed by flow cytometry. The level of the IRBP-specific Th1 cells was significantly increased and kinetically changed during EAU induction, but the cells reached peak time early before the disease was onset. Those IRBP-specific Th1 cells in the PBLs were evidence for EAU disease, but its peak time was different from EAU disease in the eyes. Our data suggested that it is very important to collect blood from patients at a suitable time point and the Th1 cells measured by flow cytometry are good marker for disease diagnosis.

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Materials and methods

Animals. Female 6–8-week-old B10.A mice were purchased from Jackson laboratory (Bar Harbor, ME) and were maintained in plastic cages under specific pathogen-free facility of Chinese Academy of Sciences (Shanghai, PR China). All animal care and use are in compliance with institutional guidelines.

EAU induction and CsA administration. B10.A mice were immunized s.c. with 50 μg of interphotoreceptor retinoid-binding protein (IRBP) in 0.2 ml emulsion (1/1 v/v) with CFA (Sigma) that had been

EAU disease developed on days 15 to 21 after immunization

In order to evaluate the time-course of EAU histopathology during disease induction, EAU score was analyzed by histopathology examination kinetically. The histological features of EAU are shown in Fig. 1. There were 5 mice in each time point. The picture in Fig. 1 shows representative results from each group. The inflammation was not observed on days 3 and 7 (Figs. 1B and C). Clinical onset of EAU in B10.A mice occurred on day 15 and disordered arrangements of the outer nuclear layer and the

Discussion

In this study, the correlation between EAU histopathology and IRBP-specific Th1 cells in PBLs was investigated kinetically during EAU induction. The significant correlation between the histopathology and Th1 cytokines in disease target-eyes was consistent with our previous work [8]. It was interesting to observe that the number of Th1 cells (CD4+ and IFN-γ+ T cell) in the peripheral blood detected by flow cytometry was increased during disease induction, but it reached a peak before disease

Acknowledgements

We are grateful to Dr. Ye Ming for reviewing the manuscript and for helpful comments. This work was supported by 973 project (G1999053907), National Natural Science Foundation of China (30070708), and key Projects of Chinese Academy of Sciences (KSCX2-2-08-02) and National Key Basic Research Program of China (2001CB510006).

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Abbreviations: EAU, experimental autoimmune uveitis; PBLs, peripheral blood lymphocytes; IRBP, interphotoreceptor retinoid-binding protein; Th1 cell, T helper type 1 cell.

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