Combination of fetal HLA-C and maternal KIR haplotypes, in comparison with the presence of the lesion in the chorionic plate.

<p>Number.</p><p>Fisher exact (2-sided).</p

Combination of fetal HLA-C2 and maternal KIR B, in comparison with the presence of the lesion in the chorionic plate.

<p>Number (percentage).</p><p>Fisher exact (2-sided) p = 0.087.</p

Antibodies.

†<p>Tris/EDTA: 0.1 mol/L (pH 9.0).</p>*<p>Envision: Dako, North America Inc, USA (anti-mouse and anti-rabbit Envision HRP).</p

Patient characteristics of the two ED groups of pregnancies, those with and without the lesion in the chorionic plate.

<p>Average (range).</p>*<p> <b>p<0.05.</b></p

Significant relationship between the presence of the lesion in the chorionic plate and histological parameters.

<p>A. Bar graph of placental histological scores in percentage. B. Incidence of histological parameters in placentas with and without the lesion in the chorionic plate. A significant relationship was found between the presence of the lesion in the chorionic plate and intervillositis and histological parameters in the decidua; including chronic deciduitis, presence of plasma cells in the decidua and fibrin deposition in the decidua. Number (percentage) Fisher's exact test. * p<0.05 ** p<0.01.</p

Higher expression of the macrophage marker CD14+, the M2 marker CD163+ and DC-Sign in the lesion.

<p>Immunohistochemical stainings of egg donation placentas. In the left panel serial slides with no lesion present and in the right panel serial slides with the lesion present in the chorionic plate. From top to bottom are depicted the stainings H&E, Cytokeratin-7, CD56+, CD8+, CD14+ and CD163+. We observed a higher expression of the macrophage marker CD14+ and the M2 marker CD163+.</p

Data behind all figures.

This file contains all data points for all figures in the associated PLOS ONE manuscrip

No difference in half-life of IgG2 light chain isotypes κ and λ in mice.

<p>(A) Recombinant human IgG2κ and λ in Balb/C mice was injected and measured by total IgG ELISA for a two week period following injection of 200 µg IgG. Calculated half-lives were 7.2±1.48 and 6.4±0.84 days for IgG2κ and IgG2λ. (B) Enrichment of IgG2 κ isoforms was performed as described in Dillon et al 2008. HPLC elution profiles of IgG2 κA and κB structural isomeres on a Dionex ProPac WCX-10 (4.0_250 mm) column are depicted. IgG2κB isoform was generated by incubation of 3 mg/ml IgG2κ in 200 mM Tris buffer at pH 8 with 6 and 1 mM of cysteine and cystamine, respectively. For IgG2κA synthesis 0.9M guanidine hydrochloride (GuHCl) was also added. The samples were kept in the dark and placed at 4°C for 48–72 h. Following incubation the antibody was run on a Zeba spin desalting column (Pierce) for buffer exchange into PBS. (C) The clearance of IgG2λ, IgG2κA, and IgG2κB in Balb/C mice. Calculated half-lives were 4.0±0.58, 5.39±0.85, and 3.7±1.04 days for IgG2κA, IgG2κB and IgG2λ, respectively. (D) Clearance of IgG2κA and IgG2κB in WT and FcγR −/− C57Bl/6 mice. Calculated half-lives were 6.2±2.62, 6.43±1.69, and 7.5±1.89 days for IgG2κA, IgG2κB and IgG2λ, respectively, in WT mice but 1.08±0.28, 1.19±0.23, and 0.7±0.92 days for IgG2κA, IgG2κB and IgG2λ, respectively, in FcRn −/− mice. Graphs in (A, C–D) depict mean and standard deviations of results obtained for 4 mice per group. Half-lives were calculated assuming exponential decay and reported in days ± standard error of means. No significant difference in half-life was detected between the two isotypes.</p

Validation of IgG1- and IgG2- light chain specific ELISA.

<p>Results from IgG1 total (A) and IgG2 total (B) were plotted against the sum of IgG1κ and IgG1λ, or IgG2κ and IgG2λ, respectively. The results of regression analysis are indicated in each panel, along with Pearson's correlation.</p

Equal placental transport and serum clearance of IgG1 and 2 light chain isotypes κ and λ.

<p>(A) The average IgG1 and IgG2 placental transport (maternal/child) ratios were compared according to their light chain isotype. (B) Clearence of IgG1 and IgG2 κ and λ was investigated in humans by collecting blood from hypogammaglobulinemia patients four weeks after an IVIg transfusion. IgG1 and IgG2 light chain isotypes κ and λ were quantified in serum by subclass- and light chain specific ELISA and subclass composition was compared to that found in the IVIg used. No preferential clearance of one light chain isotype was detectable in either IgG subclass.</p