Weight indices of recipients receiving BM with or without depletion of donor-derived α/β TCR⁺ cells.

<p><i>LEW</i>.<i>1W (RT1</i>^<i>u</i>, <i>RT7</i>^<i>a</i><i>) recipients were conditioned by 15 mg/kg of anti-RT7</i>^<i>a</i><i>mAb 3 days prior to transplantation of 1 x 10</i>^<i>8</i><i>BMC from MHC syngeneic (LEW</i>.<i>1U-7B</i>: <i>RT1</i>^<i>u</i>, <i>RT7</i>^<i>b</i><i>)</i>, <i>MHC haploidentical (LEW</i>.<i>1U-7B x LEW</i>.<i>7B</i>: <i>RT1</i>^<i>u/l</i>, <i>RT7</i>^<i>b</i><i>) or MHC disparate (LEW</i>.<i>7B</i>: <i>RT1</i>^<i>l</i>, <i>RT7</i>^<i>a</i><i>) donors</i>. <i>BM grafts were depleted from α/β TCR</i>^<i>+</i><i>cells in vitro (TCD) or were kept untreated (no TCD)</i>. <i>Thus</i>, <i>1 x 10</i>^<i>8</i><i>BMC contained 5</i>.<i>4 x 10</i>^<i>6</i><i>α/β TCR</i>^<i>+</i><i>cells (MHC syngeneic)</i>, <i>5</i>.<i>1 x 10</i>^<i>6</i><i>α/β TCR</i>^<i>+</i><i>(MHC haploidentical) and 4</i>.<i>2 x 10</i>^<i>6</i><i>α/β TCR</i>^<i>+</i><i>cells (MHC disparate)</i>, <i>respectively</i>, <i>when untreated</i>. <i>Weight indices were calculated by dividing the actual weight of the recipients through the baseline weight at the time point of BMT conditioning (= anti-RT7</i>^<i>a</i><i>mAb injection)</i>. <i>Weight indices were calculated and depicted as mean values +/- standard deviation per group (</i>^<i>…</i><i>○</i>^<i>…</i><i>depletion of</i> α/β TCR⁺ cells; –<i>■– no depletion of</i> α/β TCR⁺ cells) <i>Statistical analyses were performed applying the unpaired t test (* p < 0</i>.<i>05)</i>.</p

MHC (RT1) and CD45 (RT7) immunogenetics of strain combinations used for the bone marrow transplantation model.

<p>MHC (RT1) and CD45 (RT7) immunogenetics of strain combinations used for the bone marrow transplantation model.</p

Multi-lineage chimerism in stable recipients of MHC haploidentical BM grafts conditioned by 15 mg/kg of anti-RT7^a mAb at day 200.

<p><i>Blood samples of long-term surviving recipients of MHC haploidentical BMT (F1 (LEW.7B x LEW.1U-7B: RT1^l/u RT7^b) → LEW.1W: RT1^u, RT7^a) were analysed by flow cytometry for donor-derived chimerism (RT7^b) within leukocyte lineages at day 200. Used mAbs were anti-α/β TCR mAb (R73) for T-lymphocytes, anti-CD45RA (OX33) for B-lymphocytes, anti-granulocytes (HIS48) for granulocytes and anti-RT7^b (HIS41) for donor-derived cells.</i></p

Reconstitution of donor and recipient derived leukocytes after MHC-disparate and MHC-haploidentical BMT conditioned by 15 mg/kg of the anti-RT7^a mAb.

<p><i>LEW.1W recipients (RT1^u, RT7^a, n = 6 per group) received anti-RT7^a mAb (15 mg/kg) 3 days prior to BMT. BMC of MHC disparate LEW.7B (RT1^l, RT7^b) and MHC haploidentical (LEW.1U-7B x LEW.7B (RT1^u/l, RT7^b)) donors were depleted from α/β TCR⁺ cells in vitro and intravenously injected (1x 10⁸ BMC per BMT). Numbers of surviving animals per group are indicated above the diagram. Total counts of leukocytes in peripheral blood are given. Donor- and host-derived fractions were determined by flow cytometry using a donor-specific anti-RT7^b mAb. Mean values of leukocytes from recipient origin (□) and from donor origin (■) are depicted. (A) Recipients of MHC disparate BMC lost chimerism over time, developed pancytopenia and died between days 26 and 41. (B) Recipients of MHC haploidentical BMC developed permanent chimerism and survived indefinitely.</i></p

Transplantation of BM with or without depletion of donor-derived α/β TCR⁺ cells.

<p>Transplantation of BM with or without depletion of donor-derived α/β TCR⁺ cells.</p

Biodistribution and effects of biotinylated anti-RT7^a mAb in lympho-hematopoietic compartments of RT7^a/RT7^b chimeras.

<p><i>To enable reliable flowcytometric gating of lymphocyte subsets, which will be strongly depleted by the anti-RT7^a mAb, six mixed chimeras carrying disparity only in the RT7 (CD45) antigen were generated by sublethal TBI of 7 Gy. Reconstituted rats showed a multi-lineage RT7^a-chimerism of 10 to 30 percent differing between lineages and compartments (day 0 = before mAb application). The biotinylated anti-RT7^a mAb was applied once (2 mg/kg) before the tissue compartments were analysed by flow cytometry at days 3, 7, 14, 21 and 35. The fraction of persisting RT7^a positive cells is given as percentage of all CD45⁺ cells per lineage (column height). The black column gives the fraction of RT7^a positive cells that is coated by the biotinylated anti-RT7^a mAb. T-lymphocytes (α/βTCR⁺) and NK cells (NKR-P1⁺) were strongly and compartment independently depleted, whereas B-lymphocytes (CD45RA⁺) were coated by anti-RT7^a mAb, but were not significantly reduced in cell number despite high-graded coating. (PB–peripheral blood, THY–thymus, LN–lymph node, SP–spleen, BM–bone marrow).</i></p

Effects of anti-RT7^a mAb on bone marrow level.

<p><i>A single dosage of 10 mg/kg of anti-RT7^a was injected into LEW.1W rats (n = 4). (A) BMC were analysed 7 days after mAb injection, whereby BMC were categorized by CD45 expression density and granularity using flow cytometry. Myelopoiesis showed a right shift, whereby SSC^low/CD45^low early progenitors as well as SSC^low/CD45^high lymphoid progenitors were markedly reduced. (B) Histology verified the right shift in myelopoiesis.</i></p

Patient characteristics (n = 107).

<p>BMI, body mass index; SAPS II, Simplified Acute Physiology Score II; ICU, intensive care unit; AKI, acute kidney injury; RRT, renal replacement therapy; SD, standard deviation; IGFBP7, insulin-like growth factor-binding protein 7; TIMP-2, tissue inhibitor of metalloproteinase.</p><p>Patient characteristics (n = 107).</p

Baseline characteristics of the study subgroups.

<p>Data are mean (SD) or n (%). Fluid 24 = Fluid balance in the first 24 hours after ICU admission; MAP = mean "mean arterial pressure" over the first 24 hours; Hemoglobin = mean hemoglobin level in the first 24 hours; Urine output = mean urine output ml/kg/h in first 24 hours.</p><p>Baseline characteristics of the study subgroups.</p

Multivariable logistic regression model of perioperative parameters at the time of biomarker assessment with and without the (TIMP-2)^*(IGFBP7) for risk assessment of AKI, AKI Stage 2 and 3, RRT and 28-day mortality.

<p>*Likelihood-ratio test (comparing the fit of both models)</p><p>AKI, acute kidney injury; RRT, renal replacement therapy; IGFBP7, insulin-like growth factor-binding protein 7; TIMP-2, tissue inhibitor of metalloproteinase;</p><p>MAP, mean arterial pressure; AUC, area under the curve; CI, coincidence interval; OR, odds ratio.</p><p>Multivariable logistic regression model of perioperative parameters at the time of biomarker assessment with and without the (TIMP-2)^{<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0120863#t003fn001" target="_blank">*</a>}(IGFBP7) for risk assessment of AKI, AKI Stage 2 and 3, RRT and 28-day mortality.</p