bluuebunny/crossref_metadata_2025_split huggingface dataset:
Peptides from NM23B - A Transcription Factor with NDP Kinase Activity - Are Expressed on the Surface of Leukemic Cells and Are Recognized by T-Lymphocytes.
Authors: Tschiedel Sabine , Reinhardt Annette , Ruschpler Peter , Woelfel T. , Woelfel C. , Dietger Niederwieser , Chiara Gentilini 
Published: April 2020

Abstract
Objective: During the last years a growing number of MHC-restricted antigens were recognized using autologous or HLA-matched cytotoxic T-cell lines (TCL). Molecules were isolated by HPLC or identified using cDNA expression cloning from normal or malignant target cells and found to derive from normal proteins or from mutated tumor-specific proteins. The majority of the tumor-specific peptides were derived from melanoma cells. The aim of this project was to search for immunogenic peptides on leukemia cells with the help of TCLs obtained from a stem cell donor against chronic myelogenous leukaemia (CML) recipient cells and to identify the immunogenic peptides by cDNA expression cloning.
Methods: Irradiated leukemic cells of a patient with Philadelphia-chromosome positive CML were used to stimulate PBMC of his HLA-identical related donor. TCLs were expanded to &gt;200x106 cells with antigen specific stimulation and their specificity tested in in vitro assays. Restriction of the TCLs was tested in an Elispot assay using a panel of normal and malignant autologous and allogeneic cells and mAb blocking experiments (mAb W6.32, B1.23.2, SRF8-B6, GAP-A3). Subsequently cDNA libraries from leukemic cells were established in the eukaryontic expression vector pcDNA3.1/V5HISTOPO and cDNA of 100 bacterial colonies were pooled. 293T cells were then co-transfected with the cloned HLA-cDNA of the restricting HLA and with pools of cDNA (pools of 100) obtained from the leukemic cells. Transfected cells were tested with the TCL in the Elispot assay. Positive cDNA pools were further diluted in pools of 10 colonies and tested again. Positive pools were finally diluted in single clones and screened again. cDNA from the positive single clones was sequenced with different primers.
Results: The TCL PaHe recognized bcr/abl positive PBMC, CD14+ and CD34+ subpopulations and EBV-LCL but not autologous CD4+ and CD8+ subpopulations or bcr/abl negative PHA blasts. Blocking experiments showed that the recognized peptides were restricted by HLA-A32, HLA-B44 and HLA-C03. Therefore 293T cells were transfected with HLA-A32 and with the cDNA pools of the tumor cells. After testing one thousand 100-cDNA pools and four hundred 10 cDNA pools a total of three single cDNA clones were identified, which after transfection were recognized by the TCL. All three positive cDNAs clones were sequenced and found to be identical with the sequence of NM23B. No mutations of the molecule were detected even if different primers were used.
Conclusions: Peptides from NM23B, a transcription factor for c-myc with nucleosid diphosphate kinase (NDP) activity, are recognized by TCLs in an HLA-A32 restricted way. This is the first report of NM23B as an immunogenic molecule, which might play an important role in graft-versus-tumor reactions but also in the regulation of hematopoiesis.
Reduced Intensity Cord Blood Transplantation Using Mycophenolate Mofetil and Cyclosporine for Adult Patients with Advanced Hematologic Malignancies.
Authors: Hamaki Tamae , Kajiwara Koichi , Hoshi Keiko , Kogure Katsuhiro , Harima Akane , Yamada Kazunari , Kodo Hideki , Kozai Yasuji 
Published: April 2020

Abstract
Objectives: To evaluate the feasibility of reduced-intensity unrelated cord-blood transplantation (RI-UCBT) using cyclosporine and mycofenolate mofetil as graft-versus-host disease (GVHD) prophylaxis for adult patients with advanced hematological diseases
Methods: Twenty-one patients (median age, 54; range, 30–76) with advanced hematological diseases underwent RI-UCBT at Metropolitan Fuchu Hospital between July 2002 and September 2004. Underlying disease included AML (n=7), non-Hodgkin lymphoma (n=10), Hodgkin lymphoma (n=2), and aplastic anemia (n=1). Preparative regimen comprised fludarabine (Flu) and total body irradiation (TBI), combined with busulfan (BU) or cyclophosphamide (CY). GVHD prophylaxis was cyclosporine and mycophenolate mofetil. Primary endpoint of this study was engraftment. Secondary endpoints were to describe clinical features of immune reactions following RI-CBT. Immune reactions included high-grade fever (&gt; 38 degree centigrade), elevation of serum C-reactive protein (CRP) levels, skin rash or erythema, diarrhea, hyperbilirubinemia, symptoms of central nerve systems, and body weight gains. They were classified according to their onset after transplant: pre-engraftment immune reactions, engraftment syndrome, and GvHD.
Results: Ten patients achieved primary neutrophil engraftment at a median of day 22 (range, 11–33). Five patients died of disease progression before engraftment. Primary graft failure was diagnosed in the remaining 6 patients. Among them, 3 patients received second transplant, and the other 3 patients developed autologous hematopoiesis recovery. Seven of 10 patients who achieved engraftment developed pre-engraftment immune reactions including fever, an increase in CRP, skin rash, diarrhea, and body weight gain. Similar manifestations were observed in six patients who had not achieved primary engraftment. Six patients developed engraftment syndrome. Nine patients developed Grade II to IV GVHD: skin rash (n=9), diarrhea (n=7), and hyperbilirubinemia (n=1). Seven of 10 patients who achieved engraftment survived without disease progression at a median follow-up of 17.5 months (range, 8.4–29.0).
Conclusions: This study demonstrated the feasibility of RI-UCBT in adults, indicating that acute GVHD prophylaxis with cyclosporine and mycophenolate mofetil is worth considering for further intense evaluation.
Nonmyeloablative Unrelated Donor (URD) Hematopoietic Cell Transplantation (HCT) for the Treatment of Patients (pts) with Poor-Risk, Relapsed or Refractory Multiple Myeloma.
Authors: Georges George E. , Maris Michael B. , Maloney David G. , Sandmaier Brenda M. , Sorror Mohamed L. , Shizuru Judith A. , Niederwieser Dietger W. , Agura Edward D. , Bruno Benedetto , McSweeney Peter A. , Pulsipher Michael A. , Chauncey Thomas R. , Storer Barry E. , Storb Rainer F. 
Published: April 2020

Abstract
We carried out HLA-matched URD peripheral blood stem cell (PBSC) transplant in 24 pts with poor-risk multiple myeloma after nonmyeloablative conditioning with fludarabine (90 mg/m2) and 2 Gy total body irradiation. Postgrafting immunosuppression consisted of cyclosporine and mycophenolate mofetil. The median age of the 19 men and 5 women was 53 (23 – 66) years. Conventional metaphase cytogenetics obtained in 14 pts showed Δ13 in 6 pts and complex abnormalities in 9 pts. Stage III disease was present in 83% and 17% had stage II disease. The median time from diagnosis to URD HCT was 25 (range, 8–130) months, and 96% were beyond 1st complete remission (CR) or had never achieved CR1, despite multiple lines of chemotherapy (median 4.5, range 2–10). At study entry, 17 pts (71%) had chemotherapy-refractory disease and 14 pts (58%) had failed autologous HCT. Thirteen pts had planned autologous-URD tandem HCT, while 11 proceeded directly to URD HCT. The median follow-up was 2.5 years after allografting. One pt experienced non-fatal graft rejection. The incidences of acute grades II, III and chronic graft-versus-host disease were 54%, 13% and 75%, respectively. Non-relapse mortality was 22% at 2.5 years. CRs were observed in 11 pts (46%) and partial remissions (PR) in 3 (13%). Best disease responses were seen in pts given tandem autologous-URD HCT with CR in 8 pts and PR in 2 pts (77% CR+PR rate). The estimated overall survival (OS) at 2.5 years for all 24 pts was 65% and progression-free survival (PFS) 41%. Pts receiving tandem autologous-URD HCT had superior OS and PFS, 76% and 63% (Fig. A), compared to pts proceeding directly to URD HCT, 52% and 14% (Fig. B), respectively (PFS p-value=.03). Risk factors for worse OS included pts with significant medical comorbidities (p=.03), chemotherapy-refractory disease prior to HCT (p=.03), and pts who had failed autologous HCT (p=.07). Pts who failed autologous HCT had 48% OS and 30% PFS at 2.5 years. For pts with poor-risk, relapsed or refractory multiple myeloma, cytoreductive autologous HCT followed with nonmyeloablative conditioning and URD HCT is very promising treatment with low non-relapse mortality, high complete remission rates and prolonged PFS. The results also suggest that URD HCT may provide improved graft-versus-myeloma effect compared with HLA-matched sibling HCT without an increase in the risk of non-relapse mortality.
Figure Figure
Partially HLA Matched, Non-Myeloablative Allogeneic Transplantation.
Authors: Rizzieri David A. , Koh Liang Piu , Long Gwynn D. , Gasparetto Cristina , Sullivan Keith M. , Horwitz Mitchell , Chute John , Gong Jerald Z. , Lagoo Anand S. , Niedzwiecki Donna , Lu Cong Xiao , Marshall Dawn , Dowell Jeanette M. , Chao Nelson J. 
Published: April 2020

Abstract
We have investigated the use of partially matched family member donors in combination with alemtuzumab and a non-myeloablatvie preparative regimen in an effort to expand the population of patients who may benefit from allogeneic immunotherapy.
Methods: Sixty three patients received fludarabine 30mg/sq m, cyclophosphamide 500mg/sq m IV qd x 4 days and alemtuzumab 20mg IV qd x 5 days followed by infusion of partially matched family member donor stem cells. Mycophenolate alone or in combination with cyclosporine was given for 6–8 weeks following transplantation.
Results: Patient diagnoses included lymphoma/myeloma n= 16, leukemias/MDS n=33, myelofibrosis/aplasia n=3 and metastatic solid tumours n=11. The median age was 48 (range 17–66) with a median follow up of 28 months. The donor matching was 5/6 in 8, 4/6 in 22, and 3/6 in 33 cases. The median CD34+ cell dose collected was 13.47 x 106/kg (SD= 5.10) with nearly 2 logs of T cells depletion. Engraftment occurred in 92% of patients with a median of 87% donor cells responsible for patient hematopoiesis by 6 weeks following therapy. Twenty patients also received a DLI (range 105–107 CD3+ cells/kg). Grade III–IV acute GVHD occurred in only 8/63 (13%) with 14 (22%) experiencing grade II–IV. Nine patients developed chronic GVHD or failure to thrive (15%). The transplant regimen was well tolerated with 10% TRM. In those with hematologic malignancies, only 6 (12%) started in remission, though 38 (73%) attained a CR. The most common cause of death in this group was progressive disease (33%), followed by infections (19%), and GVHD (12%). Overall 1 year survival for this high risk group of 63 patients was 26%. A subgroup had aplastic anemia/myelofibrosis or leukemia/lymphoma in second or greater CR, PR and had a median disease free and overall survival of 3 years.
Figure Figure
Phenotypic analysis of lymphocyte subsets (measured by flow), and T cell VBeta family recovery (spectratype analysis using PCR) revealed robust recovery by 6 months in those without GVHD (dark bars) compared to those with GVHD (gray bars), despite T depletion.
Figure Figure
Further, the measured T cell recovery is from peripheral expansion of residual transplanted T cells as newly educated T cells are not measurable (using TRECs analysis) for at least 1 year, if at all.
Conclusions: The results demonstrate reasonable tolerance and reliable engraftment using T cell depleted, partially matched family member donors in a non-myeloablative setting with low treatment related mortality and severe GVHD. The future challenge will to be to develop strategies to improve immune recovery to enhance immune-mediated graft-versus-tumour effect and to minimize the risk of infections.
Treatment of Myeloid and Lymphoid Malignancies with Highly Purified Alloreactive CD56+CD3− NK-Cells and Haploidentical Stem Cell Transplantation: Promising Results of a Phase I Study.
Authors: Gentilini Chiara , Hilbers Urte , Hartung Goetz , Lange Thoralf , Kliem Constanze , Hegenbart Ute , Zeis Mathias , Huppert Volker , Glass Bertram , Uhrberg Marcus , Schmitz Norbert , Thiel Eckhard , Niederwieser Dietger W. , Uharek Lutz 
Published: April 2020

Abstract
NK cell alloreactivity can mediate strong graft versus leukemia (GvL) effects following haploidentical hematopoietic stem cell transplantation (HSCT). In an attempt to further improve the antileukemic effectiveness of this approach, we have adoptively transferred high numbers of alloreactive donor NK cells during the early phase after transplantation.
Method: In a phase-II study, 10 patients (6 AML, 1 MDS, 1 HD, 1 CML, 1 ALL, median age 38 yrs, range 17–48 yrs) were transplanted in late phases of their disease (5 pts. as 2nd transplantation) and received purified NK cells from their haploidentical donors at day +2 after HSCT. Conditioning consisted of 12 Gy fTBI, Thiotepa (10mg/kg), Fludarabine (5 x 30 mg/qm) and OKT3 (day −4 to +2). Two patients received a reduced conditioning with Fludarabine and OKT3 alone. NK cells were isolated from the CD34- fraction using an automated two-step procedure of CD3+ depletion and subsequent CD56+ selection.
Results: No severe technical problems occurred and a mean of 12,1 x 10E8 (16,74 x10E6/kg, range 6,12 to 27,2 x10E6/kg) CD34+ cells was selected in high purity (95,9 %) with a very low T-cell content (mean 1,83 x10E4/kg CD3+ cells, 4,5 Log depletion). A mean of 5,7 x10E8 (6,7 x 10E6/kg) CD56+CD3− NK cells was transferred (yield 70,36%, purity 70%). The mean number of contaminating CD3+ cells was 3,2 x10E4/kg (3,6 Log depletion).No severe acute toxicity attributable to NK cell infusion was observed. Hematopoietic recovery was fast with leukocytes &gt; 1/nl between day 5 and 11. Seven patients developed early grade II GvHD of the skin which promptly resolved after CSA and steroids. One patient developed late graft rejection five months after reduced conditioning, received a 2nd graft from a different donor, engrafted but unfortunately died due to pneumonia one month after the second transplantation. One pt with CML died due to adenovirus infection at day 140. One pt. with HD in 5th CR died due to pneumonia at day 85. One patient showed a relapse of the AML three months after transplant. She received a DLI with NK cells form the donor but died due to disease progression one month after. Interestingly, leukemic cells from this patient proved to be resistant to donor NK cell mediated lysis. Five of 9 patients are alive and in CCR with a median follow up of 192 days, the two patients with the longest follow up are in very good condition and free of GvHD at day +1271 and +1019. Our data show for the first time that the early adoptive transfer of high numbers of HLA-mismatched NK cells is safe and feasible.
Characterization of CD6-Negative Suppressor Cells Facilitating HLA-Haploidentical Stem Cell Transplantation.
Authors: Bigalke Iris , Falk Christine , ter Meer Dominik , Leeping Marina , Prevalsek Dusan , Tischer Johanna , Kolb Hans-Jochem 
Published: April 2020

Abstract
HLA- haploidentical stem cell transplantation (SCT) has been shown to be a suitable alternative for treatment of hematopoietic malignancies if no matched donor is available. Haploidentical family members sharing one HLA-haplotype may differ in one or more HLA-antigens of the second haplotype. The risks are rejection of the graft and severe graft-versus-host disease (GVHD). In particular unmodified grafts may cause severe GVHD, and T-cell depleted grafts may be rejected. We have designed a protocol of unmodified marrow transplantation supplemented by G-CSF mobilized blood cells (MBC) 6 days after marrow transplantation from which CD6-positive cells are depleted. These cells facilitate engraftment and modify GVHD. Patients were conditioned with 12 Gy total body irradiation (TBI), donor leukocytes, antithymocyte globulin and cyclophosphamide, unmodified marrow was given on day 0 and CD6-depleted MBC on day 6. In a first series of 53 patients CD6 cells were depleted by indirect loading and depletion with goat-anti-mouse antibody coupled to Dynabeads. The second series comprised 22 patients whose MBC were depleted using the CliniMacs system with CD6-antibody directly bound to immunomagnetic beads.
Table I shows the median and the range () of the cell numbers transplanted after CD6 depletion comparing the two methods.
Table II shows the number of patients with acute GVHD in relation to the median of all given transplants.
Rejection of the transplant was not observed in 71 evaluable patients. GVHD &gt;= 2 occurred in 47 %, GVHD &gt;= 3 in 21% of 70 evaluable patients. The incidence of acute GVHD was lower in patients given less CD4 cells (p=0.06), it was significantly lower in patients given MBC separated by immunomagnetic beads charged with the specific antibody (p &lt; 0.02). The content of CD8-positive, NK-, B- and CD34-positive cells had no effect on the incidence of GVHD.
We conclude from these results that transfusion of CD6-depleted MBC on day 6 facilitates engraftment of marrow from HLA-haploidentical donors without T cell depletion without jeopardizing engraftment and without severe GVHD, if CD4 cells are depleted. A particular advantage is the content of NK cells and CD8 T cells maintaining a potential GVL-effect.
Tab.I CD34 x 10e6/kg CD56 x 10e6/kg CD19 x 10e6/kg CD8 x 10e6/kg CD4 x 10e6/kg Indirect labelling (N=53) 7.3 (0.4–35.8) 19.8 (4.8–65.4) 36.1(5.6–117.3) 5.8 (0.75–55) 3.9 (0.12–65.4) Direct labelling (N=22) 13.4(2.5–29.1) 17.1 (1.7–57.3) 48.7 (5.1–175) 2.0 (0.12–13.2) 0.47 (0.05–2.5) Table II aGVHD 0 aGVHD I–II aGVHD III–IV total CD4 &lt;2.29 x 10e6/kg 16 10 4 30 CD4 &gt;2.28 x 10e6/kg 11 16 11 38 CD8 &lt;4.62 x 10e6/kg 11 13 7 31 CD8 &gt;4.61 x 10e6/kg 16 13 8 37
Harvesting for “A Rainy Day”: A Retrospective Analysis of Its Cost-Effectivenes and Ethical Implications.
Authors: Ortin Miguel , Bricknell Sue , Kimber Jane , Stirrup Lynn , Hofer Dietmar , Dyer Martin J. , Hunter Ann 
Published: April 2020

Abstract
Background: Peripheral blood stem cell (PBSC) harvesting from patients with haematological malignancies in complete remission is a frequently undertaken cautionary measure. Its aim is to provide a back-up of stem cells should the patient require high dose therapy at a later date, when harvesting may prove to be more difficult. This approach has economic and ethical implications if such harvests are not regularly used. We present a retrospective study on the use of these collections in patients with haematological malignancies.
Method: We retrospectively analysed our Stem Cell Laboratory database. All collections performed between January 1980 and December 2003 were reviewed. Endpoints included number of collections per condition group and number of collections used in each group.
Results: Over 13 years there were 1665 autologous collections. 396 (23.8 %) were collections from 158 patients who did not have need of high dose therapy at the time but in whom the potential need in the future existed. The majority of collections were from patients with Non Hodgkin Lymphoma (NHL) (34%) and Multiple Myeloma (MM) (27%). Only 69 (21%) of collections have been used, the rest remain stored. Collections from MM patients were the most frequently used (34.3%), followed by Hodgkin’s Disease (HD) and acute myeloid and chronic myeloid leukaemia (AML, CML), Only 8% of the collections from NHL patients have been used, the last infusion dating from 1995.
Conclusion: The regular, systematic collection of back-up harvests for all patients in remission is common practice. For patients with MM there is good evidence that this provides increased treatment options and is of value. The low percentage of collections subsequently used in NHL does not justify the cost or resources required and this practice should be reviewed.
Total Stored Used % Used NHL 139 128 11 8 HD 19 14 5 26 MM 105 69 36 34.3 CLL 18 18 0 0 AML 41 31 10 24.4 CML 28 22 6 21.4 ALL 16 15 1 6 APML 4 4 0 0 MDS 2 2 0 0 Miscellaneous 24 24 0 0
Plasma Is a Reliable Source of mRNA for Testing IgVH Mutation Status in Patients with Chronic Lymphocytic Leukemia.
Authors: Ma W. , Jilani I. , Gorre M. , Keating M. , Chan H. , Tseng R. , Kantarjian H. , O’Brien S. , Giles F. J. , Albitar M. 
Published: April 2020

Abstract
The mutational status of the immunoglobulin heavy-chain variable-region (IgVH) gene in the leukemic cells of patients with chronic lymphocytic leukemia (CLL) is an important prognostic marker. Lack of IgVH mutation is associated with rapid disease progression and shorter survival. The assay used to determine IgVH mutation status requires specific amplification of the mRNA of the expressed clonal IgVH gene in CLL. However, in some patients with CLL or lymphocytic lymphoma, the bulk of the disease resides in the lymph nodes or bone marrow, with few circulating leukemic cells. In addition, contamination of leukemic clonal cells by reactive polyclonal plasma cells can cause difficulty in obtaining homogenous IgVH mRNA for sequencing, and may therefore lead to failed sequencing or sequencing of the wrong IgVH mRNA. We have reported that plasma is enriched with leukemia-specific DNA, RNA, and protein because of the high turnover of leukemic cells relative to non-neoplastic cells. We thus reasoned that plasma might be a more reliable source of IgVH mRNA than cells from peripheral blood or bone marrow, where mixed populations of leukemic cells and non-neoplastic lymphocytes or plasma cells could hinder sequencing. We tested the plasma from 8 patients in whom routine (cell-based) testing for IgVH mutation status failed to yield results due to a paucity of leukemic cells (&lt;10% of total cells). The plasma of all the 8 samples showed unique, easily defined amplification products of the IgVH mRNA. Sequencing of these products showed the presence of mutated IgVH in 5 patients and unmutated IgVH in 3. Testing paired plasma and cell samples from 19 patients with CLL showed identical mutation rates and family types of the expressed IgVH gene. In contrast, plasma from 20 normal individuals showed no IgVH amplification products that could be sequenced. This data suggest that plasma can be used as an alternative to cells for testing IgVH mutation status. More importantly, plasma is more reliable when few leukemic cells are in circulation and the bulk of the tumor is in the bone marrow or lymph nodes.