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

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

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

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

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

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

Analysis of donor/recipient HLA and minor H antigen typing.

<div><p>A. Query section: Minor H antigen typing data can be entered by marking the checkboxes per allele per minor H antigen. HLA typing data has the 2-digit format. Full typing for all HLA loci is not required. After submitting the query, a page with results will appear</p> <p>B. Results: The results section lists minor H antigens relevant in the particular donor-recipient combination based upon the HLA typing. In case donor and recipient are minor H antigen matched, as is the case here for HA-1, the sign “ = ” will appear between donor and recipient. In case recipient and donor are minor H antigen mismatched, the disparities are indicated by an arrow. The direction of the arrow indicates the direction of the immune responses. In this example immune reactivity can occur in the recipient-to-donor (host-versus-graft, rejection) direction for HA-2 and HA-3 and in the donor-to-recipient (graft-versus-host) direction for HA-8 and for the various H-Y antigens.</p> <p>The tissue distribution of each relevant minor H antigen is listed as “broad” or “restricted”. By clicking these terms, a detailed list of target cells expressing the minor H antigen will be displayed.</p> <p>Minor H antigens are presented by selected HLA molecules. The HLA alleles expressed by recipient and donor that are not able to present a minor H antigen are listed at the bottom of the results section. Information will appear in case a particular minor H antigen cannot be presented by a specific HLA sub-allele (4-digit format; in this example HLA- DQB1*0503/0504 for DQ5/H-Y).</p></div

Structure of the minor H antigen database (dbMinor).

<div><p>Data from dbMinor are obtained via direct submission and from literature screening.</p> <p>The data in dbMinor can be analyzed via an HLA-based query, a full typing query and a minor H antigen query.</p> <p>Results from these queries are linked to external data from the NCBI database.</p></div

Minor H antigen PCR-SSP.

<div><p>PCR-SSP of DNA of two unrelated individuals (CAN and 80821) was performed and analyzed as described in the Materials and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000042#s3" target="_blank">Methods</a> section.</p> <p>The first lane of each minor H locus represents the immunogenic allele; the second lane displays the non-immunogenic allele.</p> <p>Bands relevant for minor H antigen typing are boxed.</p> <p>The products of 439 and 504 bp reflect the internal control for the immunogenic and the non-immunogenic allele respectively.</p> <p>The control band for UGT2B17 has a size of 761 bp.</p> <p>Since the minor H antigens UGT2B17 and H-Y have no allelic counterpart, the latter typing was performed with a single set of primers.</p></div

Image_1_Dissecting the impact of molecular T-cell HLA mismatches in kidney transplant failure: A retrospective cohort study.jpg

IntroductionKidney transplantation is the optimal treatment in end-stage kidney disease, but de-novo donor specific antibody development continues to negatively impact patients undergoing kidney transplantation. One of the recent advances in solid organ transplantation has been the definition of molecular mismatching between donors and recipients’ Human Leukocyte Antigens (HLA). While not fully integrated in standard clinical care, cumulative molecular mismatch at the level of eplets (EMM) as well as the PIRCHE-II score have shown promise in predicting transplant outcomes. In this manuscript, we sought to study whether certain T-cell molecular mismatches (TcEMM) were highly predictive of death-censored graft failure (DCGF).MethodsWe studied a retrospective cohort of kidney donor:recipient pairs from the Scientific Registry of Transplant Recipients (2000-2015). Allele level HLA-A, B, C, DRB1 and DQB1 types were imputed from serologic types using the NMDP algorithm. TcEMMs were then estimated using the PIRCHE-II algorithm. Multivariable Accelerated Failure Time (AFT) models assessed the association between each TcEMM and DCGF. To discriminate between TcEMMs most predictive of DCGF, we fit multivariable Lasso penalized regression models. We identified co-expressed TcEMMs using weighted correlation network analysis (WGCNA). Finally, we conducted sensitivity analyses to address PIRCHE and IMGT/HLA version updates.ResultsA total of 118,309 donor:recipient pairs meeting the eligibility criteria were studied. When applying the PIRCHE-II algorithm, we identified 1,935 distinct TcEMMs at the population level. A total of 218 of the observed TcEMM were independently associated with DCGF by AFT models. The Lasso penalized regression model with post selection inference identified a smaller subset of 86 TcEMMs (56 and 30 TcEMM derived from HLA Class I and II, respectively) to be highly predictive of DCGF. Of the observed TcEMM, 38.14% appeared as profiles of highly co-expressed TcEMMs. In addition, sensitivity analyses identified that the selected TcEMM were congruent across IMGT/HLA versions.ConclusionIn this study, we identified subsets of TcEMMs highly predictive of DCGF and profiles of co-expressed mismatches. Experimental verification of these TcEMMs determining immune responses and how they may interact with EMM as predictors of transplant outcomes would justify their consideration in organ allocation schemes and for modifying immunosuppression regimens.</p