J Korean Med Sci.  2009 Oct;24(5):853-859. 10.3346/jkms.2009.24.5.853.

Differences in Circulating Dendritic Cell Subtypes in Pregnant Women, Cord Blood and Healthy Adult Women

• Shin S
• Jang JY
• Roh EY
• Yoon JH
• Kim JS
• Han KS
• Kim S
• Yun Y
• Choi YS
• Choi JD
• Kim SH
• Kim SJ
• Song EY

• KMID: 1782006
• DOI: http://doi.org/10.3346/jkms.2009.24.5.853

Abstract

Different subtypes of dendritic cells (DC) influence the differentiation of naive T lymphocytes into T helper type 1 (Th1) and Th2 effector cells. We evaluated the percentages of DC subtypes in peripheral blood from pregnant women (maternal blood) and their cord blood compared to the peripheral blood of healthy non pregnant women (control). Circulating DC were identified by flow cytometry as lineage (CD3, CD14, CD16, CD19, CD20, and CD56)-negative and HLA-DR-positive cells. Subtypes of DC were further characterized as myeloid DC (CD11c+/CD123+/-), lymphoid DC (CD11c-/CD123+++) and less differentiated DC (CD11c-/CD123+/-). The frequency of DC out of all nucleated cells was significantly lower in maternal blood than in control (P<0.001). The ratio of myeloid DC/lymphoid DC was significantly higher in maternal blood than in control (P<0.01). HLA-DR expressions of myeloid DC as mean fluorescence intensity (MFI) were significantly less in maternal blood and in cord blood than in control (P<0.001, respectively). The DC differentiation factors, TNF-alpha and GM-CSF, released from mononuclear cells after lipopolysaccharide stimulation were significantly lower in maternal blood than in control (P<0.01). The distribution of DC subtypes was different in maternal and cord blood from those of non-pregnant women. Their role during pregnancy remains to be determined.

Keyword

Cord Blood; Dendritic Cells; Flow Cytometry; Pregnancy

MeSH Terms

Adult
Cell Differentiation
Dendritic Cells/*classification/cytology/immunology
Female
Fetal Blood/cytology/*immunology
Flow Cytometry
Granulocyte-Macrophage Colony-Stimulating Factor/metabolism
HLA-DR Antigens/metabolism
Humans
Lipopolysaccharides/pharmacology
Lymphocyte Activation
Pregnancy
T-Lymphocytes/cytology/immunology
Th1 Cells/cytology/immunology
Th2 Cells/cytology/immunology
Tumor Necrosis Factor-alpha/metabolism

Figure

• 

  Fig. 1 Identification of circulating dendritic cells (DC) by flow cytometry. (A) The mononuclear cell analysis gate was applied to light scatters (P1). (B) The gated events were analyzed for lineage cocktail (a mixture of FITC-conjugated anti-CD3, anti-CD14, anti-CD16, anti-CD19, anti-CD20 and anti-CD56 antibodies) and HLA-DR. P2 gate represented total DC (lineage-/HLA-DR+). (C) The P2-gated events were analyzed for CD123 and CD11c staining. Myeloid DC (lineage-/HLA-DR+/CD11c+/CD123±) were represented in gate P3, lymphoid DC (lineage-/ HLA-DR+/CD11c-/CD123+++) in gate P5, and less differentiated DC (lineage-/HLA-DR+/CD11c-/CD123±) in gate P4. Healthy, non-pregnant women (control), left panel; peripheral blood of pregnant women (maternal blood), middle panel; cord blood, right panel.

• 

  Fig. 2 (A) The percentage of myeloid dendritic cells (DC), lymphoid DC and less differentiated DC in peripheral blood of healthy, non-pregnant women (control), peripheral blood of pregnant women (maternal blood) and cord blood. (B) The expression of HLA-DR as mean fluorescence intensity (MFI) in subtypes of DC of each group. Error bars represent standard deviations. Differences between peripheral blood of healthy, non-pregnant women (control) and pregnant women (maternal blood) and cord blood were tested by Student's t t-test. Differences between peripheral blood of pregnant women (maternal blood) and cord blood were tested by Student's test for paired samples. *P<0.05; †P<0.01; ‡P<0.001.

• 

  Fig. 3 The scatter plot shows ratio of myeloid dendrite cell (DC)/lymphoid DC in peripheral blood of healthy, non-pregnant women (control), peripheral blood of pregnant women (maternal blood) and cord blood. Horizontal bars represent the mean ratio of myeloid DC/lymphoid DC. Differences between peripheral blood of healthy, non-pregnant women (control) and peripheral blood of pregnant women (maternal blood) or cord blood were tested by Student's t test. Differences between peripheral blood of pregnant women (maternal blood) and cord blood were tested by Student's t test for paired samples. *P<0.01.

• 

  Fig. 4 The TNF-α (panel A) and GM-CSF (panel B) secretion of mononuclear cells with or without lipopolysaccharide (LPS, 20 ng/mL) stimulation in peripheral blood of healthy, non-pregnant women (control), peripheral blood of pregnant women (maternal blood) and cord blood. Error bars represent standard deviations. Differences between peripheral blood of healthy, non-pregnant women (control) and pregnant women (maternal blood) and cord blood were tested by Mann-Whitney U test. Differences between maternal blood and cord blood were tested by Wilcoxon signed rank test. *P<0.01

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