bluuebunny/crossref_metadata_2025_split huggingface dataset:
Chemokine Profiling in Patients with Graft Versus Host Disease (GVHD) Following Allogeneic Blood Stem Cell Transplantation.
Authors: Ziegler Judy A. , Lokshin Anna , Lentzsch Suzanne , Agha Mounzer , Arnold Renate , Mapara Markus Y. 
Published: October 2019

Abstract
Development of GVHD following allogeneic blood stem cell or bone marrow transplantation (BMT) is dependent on the immunogenetic disparity between donor and host, subsequent alloactivation and the inflammatory reaction occurring secondary to conditioning regimen-induced tissue damage. The inflammatory response occurring early following conditioning therapy is thought to be particularly crucial for the subsequent development of GVHD as it occurs in response to tissue damage and is amplified by the alloresponse. Chemokines are predominantly small molecules (8–14 kd) that bind to a family of heterotrimeric G-protein-coupled receptors with a seven-transmembrane-spanning serpentine structure and play an important role in leukocyte trafficking and are thought to be involved in tissue infiltration during GVHD. Reports from several investigators using murine model have demonstrated the relevance of distinct chemokines in the pathogenesis of GVHD. Furthermore, we have recently demonstrated that conditioning intensity and genetic host factors influence the tissue and systemic expression of chemokines during the early course of GVHD in rodent BMT models. However, the pathobiologic relevance of chemokines expression at later time points during GVHD and in human GVHD is so far unclear.
Using luminex multiplex assays we studied the sera of patients, who had undergone peripheral blood stem transplantation from an allogeneic HLA matched sibling or matched unrelated donor in order to determine the chemokine profiles during GVHD. In addition to the most relevant cytokines/cytokine receptors (IL-1a, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IFN-a, IFN-g, GM-CSF, IL-12 (p40), IL-13, IL-15, IL17, TNF-alpha, TNF-RI, TNF-RII, DR5, FGF-b, VEGF, HGF, IL-1Ra, sIL-2R and sIL-6R), we analyzed the serum expression of CCL2, CCL3, CCL4, CCL5, CCL11, CXCL9, and CXCL10. Patients with acute GVHD of the gut (n=8), patients without GVHD (n = 4) and patients with chronic GVHD (n=4) were studied. Patients with acute/chronic GVHD had significantly elevated levels of CCL3 (90 pg/ml vs. 58 pg/ml, p= 0.004) and CCL11 (167pg/ml vs. 93pg/ml) compared to controls patients without GVHD. Patients with acute GVHD of the gut had also markedly elevated CXCL10, CCL4 levels although not reaching statistical significance. In addition, patients with acute or chronic GVHD were found to have significantly elevated serum levels of HGF, IL-1Ra, and TNF-RI.
In conclusion we are able to demonstrate that CCL3 appears to be the most prominent chemokine to be elevated in the serum of patients with a/cGVHD supporting results obtained in preclinical BMT models. Given the complex and frequently contradictory preclinical results, further studies are warranted to confirm our findings in larger cohorts of patients and to delineate whether targeting CCL3 or one of its receptor (CCR1 or CCR5) might prove beneficial for the treatment of GVHD.
Early Reconstitution of Natural Killer Cells Following Non-Myeloablative Hematopoietic Stem Cell Transplantation Predicts the Development of Acute Graft-Versus-Host Disease.
Authors: Boyiadzis Michael , Hakim Frances T. , Memon Sarfraz A. , Dean Robert , Fowler Daniel , Gress Ronald E. , Bishop Michael R. 
Published: October 2019

Abstract
Natural killer (NK) cells are among the first lymphocytes to recover following allogeneic hematopoietic stem cell transplantation (HSCT). Their levels may reflect both homeostatic and inflammatory processes. In this study we analyzed the early reconstitution of NK cells in fifty-one patients with hematologic malignancies after a reduced-intensity regimen consisting of fludarabine and cyclophosphamide and its relationship to the development of acute graft-versus-host disease (aGVHD) and cytomegalovirus (CMV) reactivation.
NK cells were severely depleted by the fludarabine-cyclophosphamide conditioning regimen. On the day of the transplant, the NK levels had been reduced in all patients to 0 cells/microliter. Subsequent NK cell recovery was rapid. By two weeks post-transplant NK cells had recovered to median levels comparable those in the original hosts (median= 119 cells/microliter; range, 0–816) and by four weeks increased to a median of 161 cell/microliter (range, 6–701; p< 0.0001) as compared to pre-transplant levels. By the second month post-transplant the median levels decreased and were maintained at pre-transplant levels.
CMV antigenemia developed in 23 of the 51 patients at a median of 30 days post transplant (range, 20–213 days). The median NK cell reconstitution at 2 weeks in the patients who subsequently developed CMV antigenemia was 158 cells/microliter (range, 2–816) and was higher than that among patients without CMV antigenemia (median 71 NK cells/microliter, range 10–228;Wilcoxon rank sum test p = 0.05).
Grade I to III aGVHD developed in 31 out of 51 patients (median time = 27 days; range, 11–92 days). The NK cell levels in the patients who developed aGVHD were significantly higher at two weeks post-transplant (median NK = 193 cells/microliter; range, 0–816) than those in the patients who did not (median NK = 66 cells/microliter; range 2–215) (p = 0.005 by Wilcoxon rank sum test). Furthermore, based on a Kaplan-Meier analysis of time to aGVHD as a function of the NK levels at two weeks post-transplant, patients who had NK cell counts greater than 195 cells/microliter at two weeks post transplant had a significantly higher probability of developing aGVHD (p = 0.0036 by log-rank test) Figure 1.
These results suggest that the early NK cell reconstitution may serve as a biomarker of transplant outcomes, especially in predicting the development of acute GVHD.
Figure Figure
Transplantation of Positive Selected Peripheral Stem Cells with Add-Back of T Cells from Unrelated Donors in Children: Favourable Survival and Low Incidence of GvHD.
Authors: Lang Peter J. , Greil Johann , Bader Peter , Mura Ulrike , Schumm Michael , Feuchtinger Tobias , Pfeiffer Matthias , Handgretinger Rupert 
Published: October 2019

Abstract
There is still a significant risk of GvHD in transplantation of peripheral stem cells from matched unrelated donors, despite the use of ATG and pharmacological prophylaxis. Especially in the case of young pediatric patients and adult donors, grafts with high numbers of stem cells but also with extreme amounts of T cells can be obtained. We present an update of a clinical study with T cell reduced grafts in 22 patients, which offer both large stem cell numbers and clearly defined amounts of T cells. Positive selection of peripheral stem cells was done with CD34 or CD133 coated magnetic microbeads and the CliniMACS™ device. Afterwards, an unselected aliquot containing 10x106/kg T cells was added to the purified stem cells and infused on day 0, resulting in a median T cell depletion of 2 log. The diagnoses were: acute lymphatic leukemia (n=9; CR1=4, CR2=5), NHL (n=1) acute (n=5; CR1=1, CR2=4) and chronic myeloic leukemias (n=3), MDS (n=3) and Wiskott-Aldrich (n=1). Median age was 10 years (0.5–18). The donors were matched for HLA-A/B (medium resolution typing) and DRB1/DQB1 (high resolution). A short course of MTX (2–3x10 mg/m2) and CSA (2–3 mg/kg, adjusted to blood levels) until day 100 were given. 21/22 patients had primary engraftment with a median time to ANC >500 of 18 days (12–60). No G-CSF was given. One patient rejected his graft and was successfully retransplanted from another MUD. Platelet recovery was fast (median time to reach independence from substitution: 20 days). Mean numbers of CD3+, CD3+CD4+ and CD3+CD8+ on day 180 (365) were 259/μl (868/μl), 100/μl (400/μl) and 146/μl (456/μl). 19/22 patients had GvHD grade 0–I (86%), 1 and 2 patients had grade II and III, (5%, 10%) respectively. Chronic GvHD occurred in 2 patients (10%). 15/22 patients are alive with a median follow up of 3 years (1.5–5). 3 year EFS was 64% (all patients), 50% (ALL/NHL) and 73% (myeloic leukemias/MDS). Relapse probability at 2 years was 25%, probability of TRM at 2 years was 15%.Causes of death were relapse (n=4) and infections (n=3). Thus, stable and favourable survival rates with a low incidence of GvHD were achieved. The method allows to administer clearly defined T cell numbers independent from the size of the grafts. This may be of advantage in particular in small children and if the donor does not accept a bone marrow harvest: all available stem cells can be infused without the limitation of unacceptable high T cell numbers.
Tacrolimus Concentrations in Hematopoetic Stem Cell Transplantation Patients Are Related to Cytochrome P450 3A5 Gene Polymorphisms.
Authors: Kobayashi Koichiro , Ogasawara Masahiro , Kikuchi Misato , Kimura Satoru , Miyazono Takayoshi , Nakata Masanobu , Imai Kiyotoshi , Hirano Teiichi , Kobayashi Naoki , Kiyama Yoshio , Tanaka Junji , Imamura Masahiro , Tanimoto Mitsune , Kasai Masaharu 
Published: October 2019

Abstract
Objective: Tacrolimus, a commonly used potent immunosuppressive drug in hematopoetic stem cell transplantation (SCT), has large individual variation in concentrations and a relatively narrow therapeutic range. It is metabolized mainly by cytochrome P450 (CYP) 3A which has genetic polymorphisms influencing enzymatic activities; CYP3A5*1 and *3 genotypes represent expressor and non-expressor, respectively. In the present study, we investigated whether the tacrolimus concentration per dose is associated with CYP3A5 gene polymorphisms.
Methods: 51 Japanese SCT patients (21 AML, 12 ALL, 9 CML, 3 NHL, 1 HL, 2 ATL, 1 MM, 2 AA), receiving tacrolimus were studied. DNA was extracted from peripheral blood samples. CYP3A5 genotype was analyzed by PCR-restriction fragment length polymorphism (RFLP) using specific primers and a restriction enzyme, SspI. Blood tacrolimus concentrations were measured twice a week by a commercially available EIA kit.
Results: The frequency of the CYP3A5 *1/*1, *1/*3 and *3/*3 genotype was 7.8, 31.4, and 60.8% of 51 patients, respectively, and the genotype distribution was not different significantly from that of 121 Japanese healthy volunteers (2.5, 38.8, and 58.8%, respectively). The frequency of CYP3A5 genotypes of white Americans is reported to be 2.2, 12.0 and 86.0%, respectively, indicating the ethnic difference of distribution. The dose-adjusted concentrations of tacrolimus 5 to 7 days following SCT were significantly higher in CYP3A5 *3/*3 patients than in CYP3A5 *1/*1 and *1/*3 patients (691.7±298.0 ng/ml per mg/kg vs. 451.8±153.8 ng/ml per mg/kg, P=0.0017). The dose needed to achieve the target concentration (10 to 15 ng/ml) between 40 to 60 days following SCT, when the blood concentrations of tacrolimus tended to be stable in most patients, is significantly lower in CYP3A5 *3/*3 patients than in CYP3A5 *1/*1 and *1/*3 patients (0.008±0.005 mg/kg vs.0.016±0.006 mg/kg, P<0.001). Above grade two adverse events related to tacrolimus were observed only in CYP3A5 *3/*3 patients (3 patients in grade 4 and 1 patient in grade2), suggesting the contribution of a higher concentration to renal toxicity by tacrolimus.
Conclusion: The tacrolimus concentration per dose in SCT patients is influenced by the CYP3A5 gene polymorphism. This is the first report showing the relationship between CYP3A5 gene polymorphism and the tacrolimus concentration in SCT patients to the best of our knowledge. Information of CYP3A5 genotype may be helpful to determine the initial tacrolimus dose adequate for each patient.
Evaluation of Pre-Transplantation T-Cell Receptor Excision Circles (TREC) in Reduced-Intensity Stem Cell Transplantation (RIST) Recipients.
Authors: Morita-Hoshi Yuriko , Heike Yuji , Yamagata Shizuka , Tajima Kinuko , Taniguchi Shuichi , Tobinai Kensei , Takaue Yoichi 
Published: October 2019

Abstract
〈Background and Methods〉: Previous studies have shown that the recipient’s thymic function before hematopoietic stem cell transplantation (HSCT) is critical for subsequent immune reconstitution and that the value of pre-transplantation T-cell receptor excision circles (TREC) is related to the incidence of graft-versus-host disease (GVHD), infection and survival. However, these observations have been mainly based on younger patients, who were eligible for myeloablative HSCT. In contrast, little is known about thymic function in elderly patients and its influence on the outcome of reduced-intensity stem cell transplantation (RIST). In this study, TREC values were measured in a total of 180 HSCT recipients who received myeloablative HSCT (n=60: mean age, 34 y) or RIST (n=120, 51 y). Quantification of thymic signal joint TREC was performed by real-time quantitative PCR and TREC values were calculated as TREC copies per 10 ng DNA obtained from peripheral blood mononuclear cells.
〈Results〉: The mean and median pre-transplantation TREC values were 3.9 and 1.3, respectively, and these showed a significant inverse correlation with patient age (p=0.0002). The mean TREC value was 3.0 in males and 5.1 in females (p=0.22), and was significantly lower in 19 patients who had received previous radiation therapy involving the mediastinum (0.96, p=0.003). The TREC value was 3.7 in 99 patients who had a disease duration of longer than 365 days, and 4.2 in 81 patients with a shorter duration (p=0.61). The TREC value was 5.5 for solid tumor (n=10), 5.0 for CML (n=10), 4.8 for AML (n=52), 3.7 for MDS (n=23), 4.0 for ALL (n=23), 2.8 for lymphoma (n=60) and 0.4 for myelofibrosis (n=2), with no difference between those in complete remission and others (both 3.9). The clinical outcome in relation to the TREC value was analyzed in 98 patients who received RIST from an HLA-identical donor. A comparison of 51 patients with lower pre-transplant TREC values (<1.3) and 47 patients with higher values (>1.3) disclosed that the incidence of acute GVHD was 64% vs 57% (p=0.52), that of chronic GVHD was 69% vs 77% (p=0.56), CMV reactivation was 72% vs 60% (p=0.20) and progression-free survival at day 100 was 72% vs 100% (p=0.0001).
〈Conclusion〉: Pre-transplant TREC values showed an inverse correlation with age, and were significantly lower among patients who received radiotherapy involving the thymus. Although these differences were not statistically significant, patients with lymphoid malignancy, patients with longer disease duration and male patients tended to have lower TREC values, while the disease status did not influence the TREC value. This study provides the baseline reference data regarding the thymic function in patients undergoing RIST, although further study is required to evaluate the significance of TREC on the clinical outcome of RIST.
Clinical Correlation of Foxp3 Regulatory Gene Expressions and NK Cells in Early Engraftment after Myeloablative Allogeneic Stem Cell Transplantation.
Authors: Yang Deok-Hwan , Park Chung-Sun , Kim Yeo-Kyeoung , Lee Je-Jung , Kim Hyeoung Joon 
Published: October 2019

Abstract
Regulatory T cells (Tregs) have been established as a key role of self tolerance and preventing proliferation of auto- and alloantigen-reactive T cells. CD4+CD25+ Tregs suppress GVHD in animal models, but in human data on GVHD following stem cell transplantation (SCT) is conflicting. We postulate that CD4+Foxp3+Tregs not only reduce the incidence of acute graft-versus host disease (aGVHD) but inhibit the NK cell functions in patients who received allogeneic SCT. CD4+Foxp3+Tregs also may adversely affect the GVL effect and cause the relapse of disease.
Patients and Methods: 25 patients (AML:13, ALL:9, CML:2, NHL:1) were undergone allogeneic SCT. Nine patients were infused from unrelated donor and five patients were from HLA one-mismatch. Three patients added alemtuzumab to myeloablative conditioning regimen due to HLA mismatch. Peripheral blood mononuclear cell (PBMC) were separated 3 or 4 weeks after SCT when absolute neutrophil count reached above 1000 × 109/L. NK cells were phenotypically analyzed by flow cytometry using directly conjugated antibodies to CD3 and CD56. CD4+ cells were isolated from PBMC using micro-bead (MACS) and the expression levels of Foxp3 mRNA were assessed by quantitative real-time PCR.
Results: 12 patients developed grade 2–4 acute GVHD and 10 patients relapsed. Patients who experienced Gr2-4 aGVHD had significantly lower the relapse rate than those who Gr0-1 aGVHD (P=0.002). Foxp3 gene expressions within CD4+ T cells were significantly lower in Gr2-4 aGVHD patients (median, 4.278 ng/μl) than in Gr0-1 aGVHD patients (median 7.914 ng/μl) except three patients treated with alemtuzumab conditioning (P=0.016). All of three patients used alemtuzumab experienced the relapse and no aGVHD. They also had very lower Foxp3 gene expressions (median, 4.760 ng/μl) than those in Gr0-1 aGVHD patients. Without alemtuzumab used patients, the levels of Foxp3 expression in relapsed patients were significantly higher than those in non-relapsed patients (median, 11.684 ng/μl and 2.031 ng/μl, respectively) (P=0.001). However, we could not find an inverse correlation between NK cell expressions and CD4+Foxp3+Tregs and a positive correlation between infused T cell doses and CD4+Foxp3+Tregs expression.
Conclusion: The levels of CD4+Foxp3+Tregs affect the incidence of aGVHD and predict the risk of the relapse. Alemtuzumab may influence the T cell recovery including regulatory T cells after early post-SCT.
Decreased Serum Levels of Surfactant Protein D (SP-D) Prior Transplant Is Associated with the Development of Bronchiolitis Obliterans (BO) and Idiopathic Pneumonia Syndrome (IPS) Following Allogeneic Hematopoietic Stem Cell Transplantation.
Authors: Nakane Takahiko , Nakamae Hirohisa , Kamoi Hiroshi , Koh Hideo , Takeoka Yasunobu , Sakamoto Erina , Kanashima Hiroshi , Ichihara Hiroyoshi , Nakamae Mika , Koh Ki-Ryang , Yamane Takahisa , Hino Masayuki 
Published: October 2019

Abstract
Non-infectious pulmonary complications which occur beyond 3 months of allogeneic hematopoietic stem cell transplantation (allo-HSCT) have become recognized as a critical problem. Above all, bronchiolitis obliterans (BO) and idiopathic pneumonia syndrome (IPS) are difficult to be cured and cause high mortality. Therefore, early identification of patients with higher risk for BO/IPS may lead to improving outcome of allo-HSCT.
Surfactant protein D (SP-D) is one of collectins mainly synthesized by alveolar type II cells. The best known function is to prevent the collapse of alveoli by decreasing surface tension at alveolar air-lipid interface, and SP-D plays an important roles as a mediator in innate immunity in the lung. In clinical practice, high serum SP-D level is used as a marker lung diseases. Recently, low production of SP-D in the alveoli has been considered as an important pathgenesis of adult respiratory distress.
We analyzed serum SP-D levels before allo-HSCT of 42 patients who received allo-HSCT in our institution between November 2001 and February 2006 and survived more than 90 days after transplant. In all patients, 5 patients had BO and 3 had IPS at a median interval of 303 and 117 days of transplantation (range; 100–452 and 95–153 days). As a result of univariate analysis, BO/IPS patients had lower serum SP-D levels prior allo-HSCT and extensive type of cGVHD except lung compared with no BO/IPS patients (p=0.06 and 0.07). KL-6 had also mildly associated with BO/IPS. On the other hand, we couldn’t find the association in sex, age, donor source, conditioning regimen, disease status, aGVHD, and FEV1.0/FVC and SP-A prior HSCT. We could not clarify the precise mechanism of the association between Decreased serum levels of SP-D and the development of BO and IPS following allo-HSCT. However, we speculate that low production of SP-D in the alveoli, which causes innate immune compromise, might contribute to extreme pulmonary alloreaction.
Mesenchymal Stem Cells for Treatment of Severe Acute Graft-Versus-Host Disease.
Authors: Le Blanc Katarina , Frassoni Francesco , Ball Lynne , Lanino Edoardo , Sundberg Berit , Lonnies Helena , Roelofs Helene , Dini Giorgio , Bacigalupo Andrea , Locatelli Franco , Fibbe Willem F. , Ringden Olle 
Published: October 2019

Abstract
Mesenchymal stem cells (MSC) from adult bone marrow have the capacity to differentiate into several mesenchymal tissues and inhibit T-cell alloreactivity in vitro. Within the EBMT MSC expansion consortium we have used MSC to treat grades III-IV acute graft-versus-host disease (GvHD) in 40 patients. The MSC dose was median 1.0 (range 0.4–9) 10^6 cells/kg body weight of the recipient. No side-effects were seen after MSC infusions. Nineteen patients received one dose, 19 patients received two doses, two patients received three and five doses respectively. MSC donors were in five cases HLA-identical sibling donors, 19 haploidentical donors and 41 third-party HLA-mismatched donors. Among the 40 patients treated for severe acute GvHD, 21 had complete responses, eight showed improvement, eight patients did not respond, two had stable disease and one patient was not evaluated due to short follow-up. Twenty patients are alive between six weeks up to 3.5 years after transplantation. Nine of these patients have extensive chronic GvHD. One patient with ALL has recurrent leukaemia and one patient has denovo AML of recipient origin.
We conclude that MSC have immunomodulatory and tissue repairing effects and should be further explored as treatment of severe acute GvHD in prospective randomized trials.