bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hematopoietic Stem Cell Transplantation (HSCT) for Benign Indications Using Plasma Depleted Umbilical Cord Blood Units (UCB) That Were Not Depleted of Red Blood Cells.
Authors: Rosenthal Joseph , Jaing Tang-Her , Nademanee Auayporn , Karanes Chatchada , Graham Michael , Chan Lee Lee , Eames Gretchen , Tan Ah Moy , Tan Poh-Lin , Wang Brian , Tseng Peggy , Chow Tiffany , Wu Tina , Lin Hai-Peng , Gjertson David , Petz Lawrence , Tan Patrick , Chow Robert 
Published: October 2019

Abstract
UCB is an attractive source for unrelated HSCT of benign indications; however, cell dosage is a critical factor for UCB HSCT. The red cell depletion (RD) and post-thaw wash techniques that are widely used incur significant nucleated cell loss; therefore, two strategies to minimize cell loss are to deplete plasma, but not the red blood cells (PD) during processing and forego post-thaw wash. A retrospective audited analysis was performed on 61 patients with benign disorders who were transplanted with 68 PD UCB units (8 double cords) with 29 thalassemias, 8 AA, 5 WAS, 5 SCID, 2 osteoporosis, 2 sickle cell disease, 2 Hemophagocytic Lymphohistiocytosis, 2 Hurler Syndrome, 1 CGD, 1 Fanconi’s Anemia, 1 Leroy I-Cell Disease, 1 Lymphohistiocytosis, 1 OIMD, and 1 Alpha Mannosidosis. Transplant characteristics: patient median age 4.25 years old (range 0.3–39); median weight 17 kg (range 5–76); male 56%; median # HLA ABDR matches of 5.0 (12–6/6; 19–5/6; 23–4/6; 5–3/6, 1–2/6); median pre-freeze TNC dose 8.1 x 107/kg; median post-thaw TNC dose as reported by TC 7.7 x 107/kg; median pre-freeze CD34 dose 3.1 x 105/kg; transplants outside of U.S.− 32 (52%); non-myeloablative − 9; 44% post-thaw washed (W), 56% infused without post-thaw wash (NW). The Kaplan-Meier estimates of 3-month ANC500 and 6-month platelet 20K and 50K engraftment are 87±5%, 83±6%, and 84±6% respectively. The median time to engraftment for ANC 500, platelet 20K, and 50K are 20 days (range 11–64), 46 days (range 13–153), and 61 days (range 21–171) respectively. No major adverse event was observed in either the W or the NW group, and the median time to engraftment for ANC 500, platelet 20K and 50K for W vs. NW were 21 vs. 19 days, 55 vs. 44.5 days, and 76 vs. 59 days respectively. The incidence of reported grade II–IV acute GVHD was 29%, and 10% had grade III–IV acute GVHD. 33% developed limited chronic GVHD, and 15% developed extensive chronic GVHD. With a median follow-up of 219 days (range 4–1402 days), the Kaplan-Meier estimates of 1-year TRM, OS and disease-free survival were 20±6%, 78±6% and 72±6% respectively. These results demonstrate that HSCT using unrelated PD UCB can be performed safely with outstanding results in patients with benign disorders, and post-thaw washing may delay engraftment of HSCT using PD UCB.
High-Dose Methotrexate (MTX) as Part of a Novel Immunosuppressive Regimen in Canine MHC-Haploidentical Hematopoietic Cell Transplantation (HCT).
Authors: Thakar Monica S. , Gooley Ted A. , Santos Erlinda B. , Storb Rainer F. , Sandmaier Brenda M. 
Published: October 2019

Abstract
In a preclinical canine model using major histocompatibility (MHC)-haploidentical littermate peripheral blood stem cell (PBSC) grafts, initial donor chimerism could be achieved after a nonmyeloablative conditioning regimen consisting of monoclonal antibody S5 (anti-CD44) from days −7 to −2, 200 cGy TBI on day 0, and MMF and cyclosporine for postgrafting immunosuppression. However, sustainable donor chimerisms could not be achieved in many of these dogs, with 25 of 44 (57%) rejecting their grafts at a median of 7 weeks after transplant. In order to improve the engraftment rate, a novel strategy to overcome the immunological barrier created by this MHC-haploidentical setting was developed. By adding single, progressively higher doses of MTX on day+3 after transplantation to the above regimen, we hoped to control host T and NK cells, both of which contribute to graft rejection in MHC-haploidentical HCT, and to target alloreactive donor T cells which are responsible for graft-versus-host disease (GVHD). Once dose toxicity is established, this system will also allow the study of transplants using gene-modified, MTX-resistant CD34 selected PBSC, which can provide additional intrinsic graft protection on exposure to MTX. To date, 11 dogs have been transplanted using the above regimen and adding either 50 (n=5), 100 (n=3), or 200 (n=3) mg/m2 MTX on day +3 followed by leucovorin rescue on days +4 and +5. Early results showed that 4 of 5 dogs experienced graft rejection in the MTX50 group, 0 of 3 dogs rejected in MTX100 (p=0.047, Monte Carlo), and all 3 dogs in MTX200 have continued to exhibit stable mixed chimerisms from +42 to +76 days after transplant. Dogs in all 3 cohorts had rapid engraftment within 1 week, with similar average peak mononuclear cell (MNC) chimerisms at comparable timepoints (83% at 5 wks, 88% at 6 wks, and 80% at 5 wks at MTX50, 100, and 200 respectively). None of the 11 dogs have shown evidence of GVHD or MTX toxicity, and all dogs cleared MTX within 48 hours. All dogs exhibited early donor tolerance on mixed lymphocyte culture assays within 2 weeks and preserved NK activity posttransplant. While these data are preliminary, adding single, progressively larger doses of MTX on day +3 to a novel, nonmyeloablative conditioning regimen appeared to improve engraftment among MHC-haploidentical canine recipients with no signs of GVHD. There was high initial donor chimerism without undue early toxicity. Further, this study will aid in our establishment of high-dose, posttransplant MTX as a way of promoting sustained donor chimerism after transplanting CD34 selected, MTX-resistant canine PBSC in the MHC-haploidentical setting. This preclinical data serve as a basis for developing nonmyeloablative transplant options for patients without HLA-identical donors.
Oral Administration of PBI-1402 Significantly Reduces Erythropenia and Accelerates Peripheral Blood Recovery in Myeloablated Mice Post Transplantation.
Authors: Morin Marie-Josée , Krosl Gorazd , Lagraoui Mouna , Grouix Brigitte , Penney Christopher , Gagnon Lyne , Roy Denis-Claude 
Published: October 2019

Abstract
The goal of this study was to determine the potential of PBI-1402, a compound that was shown to provide radio- and chemoprotection, to ameliorate the myeloablative therapy induced cytopenia and/or accelerate blood cell recovery of myeloablated hosts following autologous bone marrow transplantation in mouse model. Mice were exposed to irradiation and transplanted the following day with 5×105 bone marrow cells from syngeneic donors. Following transplantation, mice were divided into two groups that received two different doses of PBI-1402 orally and a control group that received PBS for 14 days post transplantation. The results showed that PBI-1402 treatment significantly diminished post-transplant erythropenia in a dose dependent manner. Similarly, determination of haemoglobin (Hb) demonstrated a significant attenuation of Hb loss and additionally, significantly accelerated post-transplantation recovery with the highest dose of PBI-1402. A significantly higher number of platelets was observed with the highest dose of PBI-1402. In addition, similar results were obtained with analysis of white blood cell (WBC) recovery. To determine if the observed effect of PBI-1402 treatment on red blood cell cytopenia and Hb recovery is the consequence of increased erythrocyte progenitor response to Epo, we performed an Epo dose response experiment on CFU-E bone marrow samples obtained from treated and control mice at different time points post transplantation. The results indicated that erythroid progenitors from treated and control mice responded similarly to Epo. However, PBI-1402 treated mice contained significantly higher number of CFU-E. Interestingly, analysis of more primitive Sca1+ progenitors revealed a significant increase in the number of Sca1+ cells from the bone marrow of mice treated with PBI-1402 (p < 0.05). These results demonstrate that PBI-1402 treatment significantly reduced irradiation induced erythropenia with concomitant acceleration in erythroid and Hb recovery. Additionally, treatment with PBI-1402 accelerated WBC recovery and platelet recovery and increased more primitive Sca1+ progenitors. PBI-1402 may offer protection from myeloabaltive therapy or accelerate recovery from remaining resident or transplanted stem cells.
Establishment of Chimerism in Rhesus Macaques through Nonmyeloablative Hematopoietic Stem Cell Transplant and Costimulation Blockade-Based Immunosuppression.
Authors: Kean Leslie , Adams Andrew , Strobert Elizabeth , Pearson Thomas , Larsen Christian 
Published: October 2019

Abstract
We established a strategy for producing high level hematopoietic chimerism after non-myeloablative conditioning in the Rhesus macaque. This strategy relies on hematopoietic stem cell transplantation after induction with a non-myeloablative dose of busulfan and blockade of the IL2-receptor in the setting of mTOR inhibition with rapamycin and combined CD28/CD154 costimulation blockade. Hematopoietic stem cells derived from bone marrow and leukopheresis products both were found to be successful in inducing high level chimerism. Mean peripheral blood peak donor chimerism was 81% with a median chimerism duration of 145 days. This prolonged chimerism occurred in the setting of full MHC disparity in unrelated transplant donors and recipients. Additional immune modulation strategies such as pre-transplant CD8 depletion, donor-specific transfusion, recipient thymectomy or peri-transplant deoxyspergualin treatment did not improve the level or durability of chimerism. Recipient immunologic assessment suggested that chimerism occurred amidst donor-specific down-regulation of alloreactive T cells, and the reappearance of vigorous T-mediated alloreactivity accompanied rejection of the transplants. Furthermore, viral reactivation constituted a significant transplant-related toxicity and may have negatively impacted the ability to achieve indefinite survival of transplanted stem cells. Nevertheless, this T cell costimulation blockade-based chimerism-induction regimen induced the longest-lived stem cell chimerism reported to date for non-human primates and thus represents an important platform upon which to evaluate emerging tolerance-induction strategies. Future nonmyeloablative transplant strategies based on immunomodulation via T cell costimulation blockade may have significant clinical relevence for nonmalignant hematologic diseases such as sickle cell disease and beta-thalassemia.
A Radiation-Free Immune and Epigenetics Based Conditioning Regimen for Allogeneic HCT: Combination of Anti-CD3 and Romidepsin.
Authors: Li Nainong , Zhang Chunyan , Lin Chia-Lei , McCulloch Bill , Wright John , Kirschbaum Mark , Forman Stephen , Zeng Defu 
Published: October 2019

Abstract
Hematopoietic cell transplantation (HCT) is a curative therapy for hematological malignancies as well as refractory autoimmune diseases. However, graft versus host disease (GVHD) remains a major obstacle in classical HCT, where recipients are usually conditioned with total body irradiation or high dose chemotherapy. We recently reported that donor CD8+ T cells facilitated engraftment and mediated graft versus leukemia (GVL) without causing graft versus host disease (GVHD) in young (6–8 weeks old) MHC-mismatched mouse recipients conditioned with anti-CD3 mAb (Blood 2005). Thereafter, we observed that anti-CD3 conditioning alone was not sufficient for induction of chimerism in old (>12 weeks) recipients, due to the higher percentage of residual host CD8+ T cells in the old recipients. Romidepsin (Desipeptide), a histone deacetylase inhibitor, was reported to induce apoptosis of human T cell lymphoma lines. We hypothesize that depsipeptide will induce the apoptosis of anti-CD3 activated proliferating T cells, and that conditioning with a combination of anti-CD3 and depsipeptide will markedly reduce the residual host T cells and allow donor stem cell engraftment.
To test our hypothesis, we first added Romidepsin (1.25–10 ng/ml) to cultures of T cells with or without anti-CD3 stimulation. We found that, although Romidepsin showed no effect on un-stimulated T cells, it augmented apoptosis of anti-CD3 activated T cells in a dose dependent manner, and the maximum augmentation was 5 fold. In addition, when Romidepsin (1.25–10 ng/ml) was added to a culture of mixed lymphocyte reaction (MLR), we found that it suppressed MLR in a dose dependent manner also, and the maximum suppression was greater than 98%. Second, old (> 12 weeks) BALB/c recipients were conditioned with one I.V. injection of anti-CD3 (20μg/g) and three I.P. injections (every other day) of Romidepsin at a dose of 0.4 μg/g. 7 days after anti-CD3 injection, recipients were injected with donor bone marrow cells (100×106) and CD4+- T depleted spleen (CD4−-SPL) cells (100×106). CD4−-SPL cells were injected again 7 days later. We found that, 4 weeks after HCT, 7/8 of the recipients conditioned with a combination of anti-CD3 and Romidepsin but only 1/8 of the recipients conditioned with anti-CD3 alone became chimeric. The recipients showed healthy appearance without signs of GVHD. The results are combined from two replicate experiments. This radiation free and GVHD preventive conditioning regimen may provide a novel approach for clinical HCT.
A PCR-Based Assay for the Detection of Langerhans-Cell Chimerism after Allogeneic Stem Cell Transplantation.
Authors: Meyer Ralf G. , Bakhtiar Shahrzad , Bender Klaus , Schmitt Timo , Konur Abdo , Huber Christoph , Herr Wolfgang 
Published: October 2019

Abstract
Early acute GVHD of the skin frequently occurs in patients after allogeneic hematopoietic stem cell transplantation. Although T cell depletion reduces the incidence and severity, it does not completely prevent skin GVHD. This leads to a prolonged need for immunosuppressive medication in a significant number of patients. For the induction of acute GVHD, the stimulation of donor T cells by residing host antigen presenting cells such as Langerhans cells of the skin (LCs) plays a central role. The absence of donor T cells after depletion, however, seems to hamper an early switch of LCs from host to donor origin. Therefore, the monitoring of LC chimerism is of great interest. We and others have provided evidence for a delayed switch in LC chimerism after T cell depleted reduced intensity stem cell transplantation. However, the assays used so far either are imprecise when applying low numbers of isolated cells or they depend on the detection of the Y-chromosome in skin sections of sex-mismatched transplants. In an attempt to set up a more sensitive assay of general applicability, we combined the detection of donor chimerism and tissue specific markers in a single multiplex PCR. We established PCRs for 10 different cDNA regions of constitutively expressed genes containing single nucleotide polymorphisms (SNPs). The SNP-containing products of the multiplex PCR were subsequently analyzed by the primer extension method (minisequencing) and subsequently analyzed by capillary electrophoresis. All SNP-containing cDNAs were expressed in peripheral blood mononuclear cells (PBMCs) as well as in isolated CD4- and CD8-positive T-lymphocytes, in myeloid dendritic cells, LCs, and keratinocytes. When we tested this approach on PBMCs of 10 patients and their HLA-matched sibling donors, the assay distinguished all pairs in 1 to 6 out of 10 systems. In a subsequent step, the 10plex PCR was combined with the tissue specific markers langerin for LCs and cytokeratin 10 to distinguish LCs from keratinocytes. Their expression was detected using gene-specific probes in the same minisequencing reaction used for the detection of SNPS. The resulting 12plex assay distinguished sibling donors from the patients with the same specificity and, in the same reaction, detected Langerin as well as cytokeratin 10 in purified LCs and keratinocytes, respectively. In summary, we established a sensitive assay allowing the simultaneous detection of donor chimerism together with the tissue specificity of isolated LCs that is independent of sex-mismatched donors. The addition of further tissue specific markers might allow performing chimerism studies on other tissue resident antigen presenting cells.
Crucial Role of FLT3 Ligand in Immune Reconstitution Following Bone Marrow Transplantation and High Dose Chemotherapy.
Authors: Buza-Vidas Natalija , Cheng Min , Duarte Sara , Jacobsen Sten Eirik W. , Sitnicka Ewa 
Published: October 2019

Abstract
Almost five decades after the first clinical transplantations, delayed immune reconstitution mechanisms regulating immune reconstitution post-transplantation remain to be established. While early recovery of innate immunity (myeloid and NK cells) results in reconstitution of protective immunity against bacterial pathogens, the levels of functional lymphocytes frequently remain abnormal for months or years. Low levels of B cells, immunoglobulins and CD4 T cells, predispose transplant recipients to severe bacterial and viral infections. Whereas the identification of granulocyte colony stimulating factor (G-CSF) and granulocyte/monocyte stimulating factor (GM-CSF) has been used to successfully promote clinical granulocyte recovery, no cytokine treatment has been developed to accelerate B and T cell reconstitution following myeloablation and BMT. While adult fms-like tyrosine kinase 3 (FLT3) ligand (FL) deficient (FL−/−) mice have reduced numbers of early B and T cell progenitors, they sustain close to normal levels of mature B and T cells (McKenna, H.J. et al. Blood 2000, Sitnicka, E. et al. Immunity 2002, Sitnicka, E. et al. J Exp Med 2003).
Herein, we demonstrate that adult BM cells fail to reconstitute B cell progenitors and conventional B cells in lethally irradiated FL−/− recipients, which also display delayed kinetics of T cell reconstitution. Similarly, FL is essential for B cell regeneration following chemotherapy-induced myeloablation. In contrast, fetal progenitors reconstitute B lymphopoiesis in FL−/− mice albeit at reduced levels. A critical role of FL in adult B lymphopoiesis is further substantiated by an age-progressive decline in peripheral conventional B cells in FL−/− mice, whereas fetally and early postnatally derived B1 and marginal zone B cells are sustained in a FL-independent manner. Thus, FL plays a crucial role in sustaining conventional B lymphopoiesis in adult mice, and as a consequence, our finding implicate a critical role of FL in promoting immune reconstitution following myeloablation and BM transplantation.
Alternative Splicing Due to an Intronic SNP Defines a Novel HLA-B44-Restricted-Hematopoiesis-Restricted Minor Histocompatibility Antigen.
Authors: Kawase Takakazu , Akatsuka Yoshiki , Torikai Hiroki , Morishima Satoko , Kodera Yoshihisa , Morishima Yasuo , Kuzushima Kiyotaka , Takahashi Toshitada 
Published: October 2019

Abstract
Minor histocompatibility antigens (mHAgs) with expression limited to hematopoietic cells represent attractive targets for immunotherapy to induce selective graft-versus-leukemia (GVL) reactions. Here we report the identification of a novel mHAg which is recognized by an HLA-B*4403-restricted CTL clone. Microsatellite allele image analysis of two DNA pools generated from CTL-defined mHAg positive and mHAg negative groups was performed using microsatellite markers set at 100 kb intervals within the segment initially mapped by two-point genetic linkage analysis and detailed mapping of the chromosomal recombinant points. This approach defined a 0.53 Mbp region of chromosome 18q21–22 containing 12 candidate genes potentially encoding the mHAg, although the target gene could not be identified. Subsequently, cDNA expression cloning studies demonstrated that the CTL epitope of interest was encoded by a novel allelic splice variant of XM_209104, hereafter designated as XM_209104-av. Indeed, this gene was found to lie within the region predicted by microsatellite allele image analysis. The immunogenicity of the epitope was generated by differential protein expression due to alternative splicing, which was completely controlled by one intronic single nucleotide polymorphism (SNP) located in the consensus 5′ splice site adjacent to an exon. To our knowledge, this is the first example of a mHAg controlled by a SNP located in a region other than coding sequences. Because the CTL lysed also HLA-B*4402 positive, mHAg positive B-LCLs, this novel epitope peptide can bind to not only HLA-B*4403 but also HLA-B*4402 which is a relatively common HLA-B allele in Caucasian populations. Finally, the finding that the novel XM_209104-av showed low or no expression in normal tissues including resting hematopoietic cells, but significantly higher expression in primary acute leukemia cells, especially those of myeloid lineage, suggest that this novel epitope may be an attractive therapeutic target for immunotherapy not only as a minor H antigen but also as a leukemia-associated antigen.
Serum Cytokeratin-18 Fragments as Quantitative Marker of Epithelial Apoptosis in Liver and Intestinal Graft-Versus-Host Disease.
Authors: Luft Thomas , Conzelmann Michael , Rodionova Elena , Hess Michael , Strohhaecker Ulrich , Benner Axel , Görner Martin , Hegenbart Ute , Ho Anthony D. , Dreger Peter 
Published: October 2019

Abstract
Graft-versus-host disease (GvHD) is the main complication of allogeneic stem cell transplantation (SCT). However, diagnosis and differentiation of GvHD from infectious or toxic clinical conditions as well as evaluation of GvHD response to immunosuppressive treatment remains a challenge. Since apoptosis is a common histopathological finding in GvHD, we investigated whether the caspase-cleaved neo-epitope of cytokeratin 18 fragments (CK18F) might be a serum marker for ongoing GvHD-induced target organ inflammation.
Longitudinal monitoring of serum CK18F kinetics by M30 antibody-based ELISA in 49 patients who fulfilled histopathological and/or clinical criteria diagnostic for GvHD following SCT showed that CK18F were highly elevated in hepatic and intestinal GvHD (6.3±3.8 fold and 4.6±0.8 fold, respectively, over baseline). Responses of GvHD to immunosuppressive therapy were paralleled by CK18F decreases. The extent of CK18F increases tended to correlate with serological severity of acute (r=0.96) and chronic (r=0.76) liver GvHD. Differentially relevant clinical conditions, such as non-complicated infection-related diarrhea and toxic hyperbilirubinemia were not associated with significant CK18F rises. In conclusion, CK18F monitoring provides a simple serum marker for quantitative assessment of GvHD-associated apoptotic activity in intestinal and hepatic GvHD and for distinguishing active GvHD from irreversible end organ damage. Prospective studies are warranted to demonstrate the significance of CK18F for diagnosis, differentiation, prognosis, and treatment guidance of GvHD.
Costimulatory Molecule-Targeted Immunotherapy of Chronic Graft-Versus-Host Disease.
Authors: Kim Juyang , Kim Hye J. , Kim Jiyoung , Park Keunhee , Choi Hye-Jeong , Yagita Hideo , Cho Hong R. , Kwon Byungsuk 
Published: October 2019

Abstract
Chronic graft-versus-host disease (cGVHD) is an increasingly frequent complication of allogeneic stem cell transplantation. Current therapies for cGVHD reduce symptoms but are not cures. Using the B10.D2àBalb/c (H-2d) minor histocompatibility antigen-mismatched model which reflects clinical and pathological symptoms of human cGVHD, we show that a single injection of an agonistic monoclonal antibody (mAb) to CD137, a member of tumor necrosis factor receptor superfamily, reverses skin fibrosis, ulceration and alopecia, ultimately leading to prolonged survival. The reversal is associated with markedly reduced helper type 2 CD4+ T cell (Th2) cytokines and increased production of the Th1 cytokine interferon-g (IFN-g). The Fas pathway is required for the therapeutic effect of anti-CD137 mAb on cGVHD. Taken together, these data indicate that anti-CD137 mAb has a therapeutic effect on cGVHD by removing Th2 cells mediating cGVHD. Our study demonstrate an agonistic mAb specific for a costimulatory molecule as a possible target for therapeutic intervention in cGVHD.