Histological Chorioamnionitis and Tregs Homeostasis.

<p>(<b>A</b>) Column bar graph representing means and standard deviations of CD31⁺ Treg percentage in CB with different staging of histological chorioamnionitis and no chorioamnionitis. Statistical significance between all groups (p = 0.018) was determined by one way ANOVA with Bonferroni post hoc test. (<b>B</b>) Column bar graph representing means and standard deviations of CD31⁻ Treg percentage in CB with different staging of histological chorioamnionitis and no chorioamnionitis. Statistical significance between groups (p = 0.013) was determined by one way ANOVA with Bonferroni post hoc test.</p

Outcome of predictive model for Treg populations.

<p>Outcome of predictive model for Treg populations.</p

Cell surface markers and fluorochrome labeling.

<p>Cell surface markers and fluorochrome labeling.</p

Cord blood flow cytometry analysis of regulatory T cells.

<p>Cord blood mononuclear cells were surface stained with fluorochrome labeled monoclonal antibodies followed by intranuclear staining for FOXP3, fixation and flow cytometric analysis. (<b>A</b>) Box-plot with lymphocytes gated based on size (FSC) and granularity (SSC). (<b>B</b>) Neonatal CD4⁺ T cells are selected based on CD3 and CD4 co-expression (box-plot). (<b>C</b>) Neonatal regulatory T cells (CD3⁺CD4⁺) are selected based on CD25 and FOXP3 co-expression (box-plot). (<b>D</b>) Regulatory T cells (CD3⁺CD4⁺CD25⁺FOXP3⁺) subpopulations selected by the expression of CD45RA and CD31 (CD45RA⁺CD31⁺ recent thymic emigrant, CD45RA⁺ CD31⁻ peripherally induced naïve regulatory T cell) (box-plot).</p

Maternal factors affect the percentages of Treg phenotypes in CB.

<p>(<b>A</b>) Column bar graph comparing median and confidence intervals of the percentage of CD31⁻ Tregs in infants (both term and preterm) whose mothers had received prenatal antibiotics. “Yes” indicates mother received prenatal antibiotics and “No” indicates no antibiotics were given. Statistical significance was determined by Mann-Whitney U test (*p<0.0001). (<b>B</b>) Column bar graph comparing median and confidence intervals of the percentage of CD31⁻ Tregs in infants (both term and preterm) whose mothers had received prenatal steroids. “Yes” indicates mother received prenatal steroids and “No” indicates no steroids were given. Statistical significance was determined by Mann-Whitney U test (*p<0.0001). (<b>C</b>) Column bar graph comparing median and confidence intervals of the percentage of CD31⁻ Tregs in infants (both term and preterm) whose mothers had received prenatal magnesium sulfate. “Yes” indicates mother received prenatal magnesium sulfate and “No” indicates no magnesium sulfate was given. Statistical significance was determined by Mann-Whitney U test (* p = 0.005).</p

Tregs subpopulations percentages are different between term and preterm infants.

<p>(<b>A</b>) Term CB (first box plot) and preterm CB (second box plot). Tregs were examined for expression of the surface markers CD45RA and CD31. (<b>B</b>) Vertical scatter plot comparing mean percentage of CD3⁺CD4⁺CD25⁺FOXP3⁺CD45RA⁺CD31⁺ in CB of term and preterm infants. Statistical significance was determined by independent sample T-test (*p = 0.0001). (<b>C</b>) Vertical scatter plot comparing median percentage of CD3⁺CD4⁺CD25⁺FOXP3⁺CD45RA⁺CD31⁻ in CB of term and preterm infants. Statistical significance was determined by independent sample T-test (*p = 0.0001). (<b>D</b>) Scatter plot graph showing a positive correlation between gestational age (X-axis, in weeks) and CB CD31⁺ Tregs percentage (Y-axis). Pearson's correlation test used (r =  0.54, p = 0.0001). (<b>E</b>) Scatter plot graph showing a positive correlation between gestational age (X-axis, in weeks) and CB CD31⁻ Treg percentage (Y-axis). Pearson's correlation test used (r = −0.62, p = 0.0001).</p

Increased percentage of Tregs in CB of preterm infants and correlates with gestational age.

<p>(<b>A</b>) Vertical scatter plot comparing mean percentage of Tregs (CD3⁺CD4⁺CD25⁺FOXP3⁺) in CB of term (n = 31) and preterm (n = 40) infants. Statistical significance was determined by independent sample T-test (*p = 0.015) (<b>B</b>) Scatter plot graph showing a negative correlation between gestational age (X-axis, in weeks) and CB Tregs percentage (Y-axis). Pearson's correlation test used (r = −0.414, p = 0.0001)</p

Maternal and Infant Characteristics.

<p>Maternal and Infant Characteristics.</p