[Expression of regulatory T cells and related cytokines in mice allogenic corneal transplantation rejection].
OBJECTIVE
To study the changes of CD4(+)CD25(+) and CD8(+)CD103(+) regulatory T cells and their relevant cytokines in corneal allograft transplantation in mice.


METHODS
BABL/c (H-2(d)) mice received corneal allografts from C3H/He (H-2(k)) mice were served as the experimental group. BABL/c (H-2(d)) mice received corneal from BABL/c (H-2(d)) mice were used as the control group. Corneal graft survival time was recorded pre-, 3rd day, 7th day, 4th week, 8th week post-operatively. Infiltration of inflammatory cells and corneal neovascularization were evaluated by histopathologic examination. The percentage of CD4(+)CD25(+) and CD8(+)CD103(+) T cell in peripheral blood and spleen was determined by flow cytometry. The expression of interleukin-10 (IL-10), IL-4, gamma-interferon (IFN-gamma) and IL-1beta in serum and aqueous humor was measured by enzyme-linked immunospecific assay (ELISA).


RESULTS
The graft rejection in experimental group occurred at 7 days to 4 weeks after the operation, averaged (14.79 +/- 1.02) days. The grafts in the control group remained clear within 8 weeks observation and the survival time is much longer than that of the allografts (chi(2) = 46.934, P = 0.000). Flow cytometry showed that the percentage of CD4(+)CD25(+) T cell in peripheral blood in the control group after surgery was (3.36 +/- 0.29)%, (4.09 +/- 0.44)%, (5.44 +/- 0.35)%, (5.73 +/- 0.53)% at day 3, day 7, 4th week, and 8th week, respectively, which was significantly higher than that in the experimental group [(2.50 +/- 0.39)%, (3.24 +/- 0.25)%, (4.20 +/- 0.45)%, (4.18 +/- 0.14)%; t = 3.828, 2.898, 3.780, 4.892; P < 0.05]. On the other hand, CD8(+)CD103(+) T cell in peripheral blood in the experimental group after surgery was (2.20 +/- 0.33)%, (2.79 +/- 0.57)%, (4.55 +/- 1.03)%, (4.31 +/- 0.07)% at different periods, which was much higher than that in the control group (t = 7.133, 4.876, 5.196, 19.960; P < 0.05). The changes of CD4(+)CD25(+) and CD8(+)CD103(+) T cells in the spleen was earlier than those in peripheral blood. ELISA showed the expression of IL-10 and IL-4 in the serum in experimental group after surgery was much lower than that in the control group (t = 3.203, 3.141, 3.012, 2.869 and 2.340, 6.681, 8.839, 8.574; P = 0.011, 0.012, 0.013, 0.019 and 0.053, 0.000, 0.000, 0.000). The serum levels of IFN-gamma and IL-1beta in experimental group were higher than that in the control group (t = 3.508, 3.265, 4.402, 5.539 and 3.630, 5.796, 1.728, 0.660; P = 0.006, 0.011, 0.002, 0.000 and 0.005, 0.000, 0.115, 0.524). Furthermore, the cytokine levels in the aqueous humor behaved similarly with those in the serum.


CONCLUSIONS
The down-regulation of CD4(+)CD25(+) Treg, IL-10 and IL-4 and the up-regulation of CD8(+)CD103(+) Treg, IFN-gamma and IL-1beta in mice allograft corneal rejection may play an important role in allograft rejection.
Wang
Xin
X
Department of Ophthalmology, Eye, Ear, Nose and Throat Hospital of Fudan University, Shanghai 200031, China.
LE
Qi-Hua
QH
Hong
Jia-Xu
JX
Xu
Jian-Jiang
JJ
Sun
Xing-Huai
XH
Wang
Wen-Tao
WT
Zhuang
Hong
H
D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D016039:Corneal Transplantation; D005260:Female; D006084:Graft Rejection; D007371:Interferon-gamma / Q000276:immunology; D016753:Interleukin-10 / Q000276:immunology; D053583:Interleukin-1beta / Q000276:immunology; D015847:Interleukin-4 / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D014184:Transplantation, Homologous
C556062:IL10 protein, mouse; D053583:Interleukin-1beta; D016753:Interleukin-10; D015847:Interleukin-4; D007371:Interferon-gamma