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. 2007 Jul;149(1):139-45.
doi: 10.1111/j.1365-2249.2007.03397.x. Epub 2007 Apr 25.

Proportion of peripheral blood and decidual CD4(+) CD25(bright) regulatory T cells in pre-eclampsia

Y Sasaki  1 , D Darmochwal-Kolarz, D Suzuki, M Sakai, M Ito, T Shima, A Shiozaki, J Rolinski, S Saito
Affiliations

Proportion of peripheral blood and decidual CD4(+) CD25(bright) regulatory T cells in pre-eclampsia

Y Sasaki et al. Clin Exp Immunol. 2007 Jul.

Abstract

CD4(+) CD25(bright) regulatory T (T(reg)) cells have been identified as a principle regulator of tolerance during pregnancy. In the setting of pre-eclampsia, however, little is known about the dynamics of these cells. In the current study, we determined CD4(+) CD25(bright) T(reg) cells in the peripheral blood using flow cytometry and forkhead box P3 (FoxP3(+)) cells at the placental bed using immunohistochemical staining. Peripheral blood mononuclear cells (PBMC) of 38 pre-eclamptic cases (17 cases Japanese, 21 cases Polish), 40 normal late pregnancy subjects (20 subjects Japanese, 20 subjects Polish), and 21 non-pregnant healthy controls (10 subjects Japanese, 11 subjects Polish) were included. We found the percentage of CD25(bright) cells within the CD4(+) T cell population in PBMC was reduced significantly in both Japanese and Polish pre-eclamptic cases than in normal pregnancy subjects (P < 0.001) and non-pregnant healthy controls (P < 0.001). Also, the percentage of FoxP3(+) cells within CD3(+) T cells in the placental bed biopsy samples of pre-eclamptic cases were decreased compared to those in normal pregnancy subjects. These findings suggest that a decreased number of T(reg) cells was present in pre-eclampsia, and these changes might break the maternal tolerance to the fetus.

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Figures

Fig. 1
Fig. 1
Expression of forkhead box P3 (FoxP3) in CD4+ CD25bright T cells, CD4+ CD25dim T cells and CD4+ CD25– T cell subsets in pre-eclampsia (a) and normal pregnancy (b). Lymphocytes were stained with allophycocyanin (APC)-labelled anti-CD25 monoclonal antibody (MoAb), phycoerythrin (PE)-labelled anti-CD4 MoAb and fluorescein isothiocyanate (FITC)-labelled anti-human FoxP3 MoAb. CD4+ lymphocytes were classified into CD4+ CD25bright cells, CD4+ CD25dim cells and CD4+ CD25– cells (left panels). The expression of FoxP3 and CD25 in CD4+ cells is shown in the middle panels. The percentages of FoxP3-expressing cells in CD4+ CD25bright, CD4+ CD25dim and CD4+ CD25– cells are shown in the middle panels. Most CD4+ CD25bright cells expressed FoxP3 and least CD4+ CD25dim cells and CD4+ CD25– cells expressed FoxP3.
Fig. 2
Fig. 2
Population of CD4+ CD25bright/CD4+ T cells in non-pregnant subjects, normal pregnant subjects and pre-eclamptic cases in the Japanese (○) and Polish (▵) subjects. The percentage of CD4+ CD25bright cells to CD4+ cells was comparable between Japanese non-pregnant subjects and Polish non-pregnant subjects, Japanese pre-eclamptic cases and Polish pre-eclamptic cases. The box means median ± 75% quartiles of the total of Japanese and Polish in each group. The percentage of CD4+ CD25bright cells to CD4+ cells in pregnant women was significantly higher (P < 0·0001) than that in non-pregnant women. This percentage in pre-eclamptic cases was significantly lower than that in normal pregnant women (P < 0·0001) and non-pregnant women (P < 0·0001).
Fig. 3
Fig. 3
Haematoxylin and eosin staining of placental bed biopsy samples in a normal pregnancy subject (a) and a pre-eclamptic case (e). Immunohistochemical staining of CD3 (b,f), forkhead box P3 (FoxP3) (c,g) and cytokeratin (d,h) on formalin-fixed paraffin-embedded placental biopsy samples in a normal pregnancy subject (b,c,d) and a pre-eclamptic case (f,g,h). Red arrows in (c) and (g) show FoxP3-stained cells, and FoxP3+ cells in the high-power field are shown at the upper right.
Fig. 4
Fig. 4
The numbers of CD3+ T cells (a), the population of forkhead box P3/CD4 (b) and the population of CD8/CD3 (c) at the placental bed biopsy of pre-eclamptic cases (•) and normal pregnant subjects (○). The box shows median ± 75% quartiles. CD4+ cell numbers were determined as numbers of CD3+ cells minus numbers of CD8+ cells.
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