bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD34 Positive Cells Mobilized with Chemotherapy Plus Recombinant Human Growth Hormone (rhGH) and Recombinant Human Granulocyte Colony-Stimulating Factor (rhG-CSF) Allows Sustained Engraftment in Previously Non-Mobilizer Patients.
Authors: Almici Camillo , Verardi Rosanna , Ferremi Piero , Re Alessandro , Cattaneo Chiara , Ferrari Emilio , Braga Simona , Rossi Giuseppe , Marini Mirella 
Published: April 2020

Abstract
The availability of adequate amounts of peripheral blood progenitor cells (PBPC) is a prerequisite for the feasibility of high-dose therapy program in patients with neoplastic diseases. Due to prior chemio-radiotherapy or disease related factors, 20–30% of patients fails to mobilize sufficient amounts of PBPC. Therefore, non-mobilizer patients cannot complete their therapeutic program. The aims of the present study were: 1) to evaluate the combination rhGH+rhG-CSF after chemiotherapy as a mobilization regimen in previously non-mobilizers patients; 2) to assess safety and tolerability of rhGH+rhG-CSF treatment; 3) to evaluate hematopoietic recovery after reinfusion of rhGH+rhG-CSF mobilized PBPC. After informed consent, three patients (2 male, 1 female), that failed a previous mobilization cycle, were remobilized with chemotherapy plus rhGH (100μg/Kg/d, sc) and rhG-CSG (5μg/Kg/d, sc).
Case Disease TNC (x 108/Kg) CD34+ (x 106/Kg) CFC (x 106/Kg) LTC-IC (x 104/Kg) 1 LNH 8.5 7.6 5.68 15.9 2 LNH 18 6.2 3.57 5.0 3 MM 15 1.3 1.26 12.0
No specific side effects were noticed due to rhGH treatment. Reinfusionof PBPC after myeloablative therapy resulted in hematopoietic recovery with 16, 9, 11 days respectively to reach an absolute neutrophil count ≥ 500/μl and 24, 15, 15 days respectively to reach a platelet count ≥ 50000/μl. Hematopoietic engraftment is complete and sustained with a follow-up of 18, 11 and 9 months, respectively.
In conclusion our data confirm that the addition of rhGH to rhG-CSF allows mobilization and collection of PBPC in previously non-mobilizer patients, and therefore the programmed high-dose therapy program can be satisfactorily completed, with sustained hematopoietic recovery.
Merit of Initiating Peripheral Blood Stem Cell (PBSC) Collections at Low Level of Circulating CD34+ Cells.
Authors: Law Ping , Tan Lip Kun , Yasir Fathalha , Soh Teck Guan , Mah Joanna K.Y. , Li Jenny , Liu Te Chih , Chen Chien Shing 
Published: April 2020

Abstract
Most patients or donors undergoing leukapheresis (LP) for autologous or allogeneic PBSC transplantation requires multiple collections to achieve a sufficient CD34+ cell dose. LP is usually initiated when peripheral blood (PB) CD34+ reached a certain level, such as 20/μl. The aim of this retrospective analysis is to summarize our institutional experience of initiating LP at a low PB CD34+ cell level of 5/μl and investigate the merit of the practice. All patients or donors underwent LP (using Cobe Spectra or Baxter Amicus) processing 3 times the blood volume. A total of 170 LP procedures (118 autologous and 52 allogeneic) was performed in 74 adult patients or donors (&gt; 40 kg) between Jan 2004 and May 2005. Autologous patients were mobilized with chemotherapy and G-CSF while allogeneic donors with G-CSF alone. A “good” LP product is defined as one having ≥ 1 x 106 CD34+ cells/kg so that a minimum dose of 3 x 106/kg can be achieved in 3 collections. Our result showed that each PBSC product contained 6.07 x 108 WBC/kg (median, range: 0.13–17.5) and 1.59 x 106 CD34+ cells/kg (0.14–24.9). Total CD34+ cells in PBSC products were correlated to PB CD34+ cell counts (r = 0.79, data not shown). As shown in Table 1, initiating LP at higher levels of PB CD34+ cell increased the proportion of good LP. Nevertheless, 76% of collections initiated at &gt; 5 CD34+ cells/μl achieved good LP criterion. It is possible that the level of PB CD34+ cells was still increasing in many patients or donors after initiation of LP at the low level. However, some patients /donors still achieved minimim CD34+ cell dose when second LP day (Day 2) PB CD34+ cell level was lower than that of first LP day (Day 1) (Table 2). These patients /donors would likely NOT have been collected if higher levels of PB CD34+ cells were used as guideline for start of LP. Eleven patients /donors whose Day 2 CD34+ cell count was below that of Day 1 achieved minimum CD34+ cell dose when LP was initiated at &lt; 20/μl. When LP was initiated at &lt; 10/μl, four individuals achieved minimum dose. All 4 were autologous patients mobilized with chemotherapy and G-CSF (3 AML and 1 NHL). In conclusion, our results showed that initiating LP at low PB CD34+ cells can be helpful to some individuals. The guideline may be especially useful in those patients that can only be mobilized marginally.
Table 1: Initiating LP at Different Levels of PB CD34+ Cells PB CD34+ Cells # LP % Good LP &gt; 5/μl 156 75.9 &gt; 10/μl 119 87.4 &gt; 20/μl 65 95.4 Table 2: Patient /Donor Achieving Dose and Levels of PB CD34+ Cells PB CD34+ Cells # Patients & Donors # Patients & Donors Achieving Dose # Patients & Donors that achieved dose when Day 2 PB CD34+ cells were lower than that of Day 1 &lt; 20/μ l 46 35 (82.6%) 11 &lt; 10/μ l 14 10 (71.4%) 4
Myeloablative Conditioning Is Well Tolerated by Older Patients Receiving T-Cell Depleted Grafts.
Authors: Novitzky Nicolas , Thomas Valda , Hale Geoff , Waldmann Herman 
Published: April 2020

Abstract
Introduction: Older age is associated with increased transplant related mortality from GvHD and organ dysfunction. Depletion of T-cells from stem cell grafts prevents GvHD and could allow more older patients to receive standard forms of myeloablative conditioning.
Patients and Methods: Adult patients with haematological malignancies received allogeneic grafts from HLA identical siblings. Conditioning was with radiation or chemotherapy based myeloablative schedules. PBPC grafts were incubated ex vivo with CAMPATH-1H (anti CD52) antibody and infused into patients. Patients were given therapeutic doses of cyclosporin until day 90 post transplantation. The survival of patients younger or older than the population median age was analysed to determine if age was a risk factor for outcome.
Results: 72 unselected patients aged from 15 to 59 years (median 44) received myeloablative conditioning followed by median of 2.64 (range 1.14–12.3) CD34+ cells exposed “ex vivo” to a median dose of 10 mg (range 7.5–50) CAMPATH-1H. There was one early death from sepsis and all the other patients engrafted. GvHD occurred in 11%. Death was procedure related in 12 (17%) and from relapse in 3. The median follow up is 539 (7–2316) days and 79% survive disease free. The diagnosis of acute leukemia and lymphoma was more common among younger and older individuals (p&lt; 0.01) respectively. 75% of 33 younger individuals and 79% (p= 0.4) in the older cohort survive disease free, at a median follow up of 911 and 475 days (p= 0.4) respectively. Furthermore, when outcome of patients younger than 40 years, aged between 40 and 49 and older than 49 years were compared, there was again no difference. Cox analysis showed that survival was not associated with diagnosis of disease and that developing GvHD was the only adverse factor for survival (p&lt; 0.04).
Conclusions: In patients receiving T-cell depleted grafts, as GvHD rates are low, myeloablative conditioning is well tolerated supporting the use of this transplantation modality up to the age of 59. This information is useful to select patients who would benefit most from reduced intensity conditioning schedules.
Potential Usefulness of Leukotriene B4 (LTB4) on Mobilization of Hematopoietic Progenitor Cells (HPC).
Authors: Mun Yeung-Chul , Lee Seung-Eun , Lee Kyung-Eun , Yoo Eun-Sun , Kang Eun Suk , Lee Sang min , Ahn Jee-Young , Bae Yun-Kyung , Kim Geun-Young , Lee Soon Nam , Chung Wha-Sun , Kim Min Kyoung , Hyun Myung Soo , Kim Jae-Hong , Seong Chu-Myong 
Published: April 2020

Abstract
The mechanisms on HPC mobilization seem to be multifactorial processes. Many cell adhesion molecules (VLA-4, ICAM-1, VCAM-1 etc), SDF-1/CXCR4, and proteases (ie, MMP-9) may be important players for this mobilization processes. However, finding more mechanisms on HPC mobilization is under intense scrutiny. So far, G-CSF is the best known cytokine for this purpose. Meanwhile, its limitation on using G-CSF is to take 4–6 days for the optimal mobilization at clinic and its side effects (ie; bone pain, high WBC counts) are not negligible. LTB4 is lipid mediator during the process of inflammation, having many roles (ie; inducer of chemotaxis, the production of nitric oxide, transepithelial migration of neutrophil). In present study, we focused on the roles of LTB4 on HPC mobilization and its feasibility on mobilization in vivo.
Samples were collected from the peripheral blood via heart puncture and the bone marrow on time dependent manner (1hr, 4hr, 6hr, 12hr, 24hr, 48hr) after LTB4 injection which was given via intravenously and the dose dependent manner of LTB4 (0.5μg, 1μg, 2μg, 3μg). These data were compared with two other groups after G-CSF injection and normal saline injection. Collected total nucleated cells were analyzed for Sca-1+/Lin- using flow cytometry and CFU studies. There were definite increase of Sca-1+/Lin- cells after 1μg of LTB4 injection group (TNC: 29.55(±0.92)×105/ml and HPC: 3.72(±0.09) %). Its control group showed as follows: TNC: 14.55(±0.21)×105/ml, HPC: 0.41(±0.04) %, (p&lt;0.05). Mobilization was optimal only after 4hours of LTB4 injection (TNC: 36.25(±2.90)×105/ml & HPC: 3.50(±0.37) %). After 4hours of LTB4 injection, there was a obvious down pattern. In terms of quantity, more than 20 fold of HPC mobilization after one LTB4 injection was observed. There were no immediate toxicity in the cohort, though long term potential side effects are under investigations. In conclusion, we showed the rapid mobilization of HPC with LTB4 at only 4 hours post-injection. This finding with LTB4 could be significant in terms of shortening the optimal period of mobilization comparing to that by G-CSF, which takes at least 4–6 days by repetitive injection to reach the optimal timing for collection. The possible synergistic effects of G-CSF with LTB4 for larger quantity of HPC and the cellular and molecular mechanism(s) of mobilization by LTB4 are being investigated in our lab.
Effect of Storage Time and Temperature on Fresh Unprocessed Cord Blood.
Authors: Vowels Marcus R. , Stylianou Jessica , Mison Leigh 
Published: April 2020

Abstract
Studies on the optimal conditions to store fresh cord blood (CB) in order to optimise haematopoietic stem cell (HSC) recovery prior to processing and cryopreservation have produced conflicting data. In this study we investigate the effects of time and temperature on CB HSC after collection. We also investigate whether the process of cryopreservation and thawing aggravates stress created by storage prior to cryopreservation. 30 CB units were collected and transported to the laboratory within 5 hours of collection. An aliquot of each CB unit was tested for nucleated cell count (NCC), CD34 count, CD34 viability and CFU-GEMM, and then frozen with 10% DMSO. Each CB unit was then separated into three 10 ml portions and incubated at 3 different temperatures (4–8°C, 21–24°C and 29–31°C). After 24, 36 or 48 hours, samples were taken and tested. An aliquot of each of the portions was then cryopreserved. After a minimum of 1 week, the cryopreserved samples were thawed and tested. For fresh samples incubated for 24 hrs or 36 hrs there was no difference over time when CB was incubated at 4–8°C, whereas, at temperatures above 20°C, there was either a trend or significant decrease in CD34 and CFU colonies with time. For thawed samples, viable CD34 and CFU colonies were decreased below baseline at either one or both 24 hr and 36 hr time points. This was particularly apparent for CFU colony measurements for samples kept at 29–31°C. At 48 hrs, at each of the temperature ranges (4–8°C, 21–24°C and 29–31°C), CFU were significantly lower for fresh (21.1, 16.8 and 14.0; baseline 31.0) and thawed (9.3, 8.5 and 3.0; baseline 2.7) samples and viable CD34 were significantly lower for thawed (2.0, 2.46 and 1.8; baseline 2.7) samples respectively (p&lt;0.001). In conclusion, the best survival /recovery was seen with CB stored at 4–8° C and for ≥ 36 hrs. Temperatures above room temperature (21–24°C) appeared detrimental. The data also indicate that this damage becomes more evident when tested after the CB has gone through the cryopreservation and thawing process, and is even more evident the higher the temperature and the greater the time of storage prior to cryopreservation. These results have implications for quality and safety of CB stored for clinical use.
Viable CD34 and CFU related to incubation time and temperature, tested pre- and post-cryopreservation. Viable CD34 cells x 10e6 CFU-GEMM /12,500 cells plated Storage temperature 4–8C° 21–24°C 29–31°C 4–8°C 21–24°C 29–31°C Baseline 3.4 3.4 3.4 20.9 20.9 20.9 Fresh 24 hrs 3.3 3.67 2.45 20.4 18.8 21.6 36 hrs 3.7 2.45 2.74 20.5 19.6 17.4 Frozen / Baseline 3.0 3.0 3.0 11.0 11.0 11.0 Thawed 24 hrs 2.7 2.8 2.5 11.8 11.8 6.1 36 hrs 2.5 2.84 2.1 10.6 10.1 4.7
Upregulation of MT1-MMP by Molecules Present in Leukaphereis Products Primes CD34+ Cell Homing.
Authors: Shirvaikar Neeta , Montano Jencet , Turner A.Robert , Ratajczak Mariusz Z. , Janowska-Wieczorek Anna 
Published: April 2020

Abstract
Hematopoietic stem/progenitor cells (HSPC) derived from mobilized peripheral blood engraft faster than those from bone marrow (BM) or cord blood (CB). We recently postulated that leukapheresis product supernatants (collected from G-CSF-mobilized patients) and their components (fibrinogen, fibronectin, thrombin, hyaluronic acid (HA) and C3 cleavage fragments) have a priming effect on HSPC homing by enhancing chemotactic responses to SDF-1 through increased incorporation of CXCR4 into membrane lipid rafts and stimulation of matrix metalloproteinase (proMMP-2 and proMMP-9) secretion in these cells (Blood2005; 105:40). As MT1-MMP has been shown to activate latent forms of MMP-2 and (indirectly) MMP-9 and is known to localize proteolytic activity at the leading edge of migrating cells, we decided to investigate MT1-MMPs’ role in the priming of HSPC homing. We observed that stimulation of BM and CB CD34+ cells with priming agents (HA, fibrinogen or thrombin) not only increased secretion of proMMP-2 and proMMP-9 (zymography) but also highly upregulated levels of MT1-MMP transcript (RT-PCR) and protein (Western blotting) in these cells. Moreover, trans-Matrigel chemoinvasion of CD34+ cells towards a low SDF-1 gradient (20 ng/mL), enhanced by priming agents, was inhibited by the potent MT1-MMP inhibitor epigallocatechin-3-gallate. Furthermore, priming agents (HA, fibrinogen and thrombin) increased levels of active MMP-2 in co-cultures of stromal cells (endothelial cells and BM fibroblasts) with CD34+ cells. To elucidate the mechanism of MT1-MMP upregulation by priming agents, we evaluated whether they affected MT1-MMP incorporation in ganglioside M1-enriched membrane lipid rafts in the same way as for CXCR4. We found that stimulation of hematopoietic and THP-1 cells with HA, fibrinogen and thrombin increased incorporation of MT1-MMP into membrane lipid rafts and these observations were confirmed using confocal microscopy. Hence it appears that MT1-MMP, like CXCR4, localizes in membrane lipid rafts, and this is enhanced by priming agents, leading to better homing of HSPC.
Higher Efficacy of Peripheral Blood Stem Cell Collection in Children Mobilized with High-Dose G-CSF.
Authors: Styczynski Jan , Debski Robert , Gornicka Hanna , Hulek Elzbieta , Wojtylak Pawel , Wysocki Mariusz 
Published: April 2020

Abstract
Objective: Analysis of efficacy of PBSC collection after mobilization by high-dose G-CSF in children and young adults, qualified for transplantation of autologous hematopoietic stem cells.
Patients and Methods: Total number of 21 patients (23 collection cycles) were included into the study, divided into 2 groups: group I of 10 patients (aged 16m-23y, weight 8.5–61kg) with solid tumors were mobilized with G-CSF (Neupogen) at dose 2x5 μg/kg bw/day (12 collection cycles); group II of 11 patients (aged 3–27y, weight 12.5–68kg) with solid tumors (n=6) or hematological malignancies (n=5), who were mobilized with G-CSF (Neupogen) at dose 2x12 μg/kg bw/day. PBSC collections were performed by COBE Spectra from the day when CD34 cell count exceeded 20/μl. Efficacy of procedure was determined by collection of total 2x106 CD34 cells/kg (minimal required cell dose), or 5x106 CD34 cells/kg (optimal cell dose).
Results: Group I: Peripheral blood CD34 cell count exceeded 20/μl in 7/12 (58%) patients at day 4 and 10/12 (83%) at day 6 of G-CSF administration. Total number of CD34 cells of 2x106 /kg kw and 5x106 /kg bw was obtained in 3/12 (25%) and 2/12 (17%) patients after first collection respectively; and in 8/12 (67%) and 4/12 (33%) after second collection, respectively. After 3 days of collection required number of cells was obtained in 9/12 (75%) and 6/12 (50%) patients, respectively. No patient reported side-effects related to G-CSF administration. Group II: Peripheral blood CD34 cell count exceeded 20/μl in 8/11 (73%) patients at day 4 and 11/11 at day 6 of G-CSF administration. Total number of CD34 cells of 2x106 /kg bw and 5x106 /kg bw was obtained in 9/11 (82%) and 3/11 (27%) patients after first collection respectively; and in 11/11 and 6/11 (55%) after second collection, respectively. After 3 days of collection, in all patients, but one, optimal number of cells was obtained. Two patients reported side-effects related to G-CSF administration (headache, bone pain). No differences were found between the rate of speed of PBSC mobilization in analyzed schemes of G-CSF administration, however group of patients mobilized with high-dose G-CSF, showed more efficient stem cell collection after 3 days of procedure (p=0.027; OR=14; 95%CI=1.1–402).
Conclusion: Mobilization of PBSC with high-dose G-CSF is a safe and effective method of hematopoietic stem cell collection, enabling high efficacy of this procedure.
Quadruple Therapy with CsA, MTX, MMF and ATG and Triple Therapy with CsA, MTX and ATG for Preventing Graft-Versus-Host Disease in Unrelated Donor Hematopoietic Stem Cell Transplantation.
Authors: Fan Zhiping , Yi Zhengshan , Liu Qifa , Sun Jing , Xu Dan , Zhang Yu , Wei Yongqiang , Ye Changxiong 
Published: April 2020

Abstract
Objective To explore the effective protocol for graft-versus-host disease (GVHD) prophylaxis in unrelated donor hematopoietic stem cell transplantation (URD-HSCT).
Methods 31 patients with leukemia received URD-HSCT, of whom 16 received quadruple therapy (quadruple group) with CsA, MTX, MMF and ATG for GVHD prophylaxis and 15 received triple therapy (triple group) with CsA, MTX and ATG. 22 patients were matched in all HLA genetic loci with donors, seven were mismatched in one HLA genetic locus, 1 in two HLA genetic loci, and 1 in three HLA genetic loci. Total body irradiation (TBI) plus cyclophosphamide (CTX) was adopted in 17 cases and modified BuCY conditioning regimen (hydroxyurea, busulfan, Ara-C, Cyclophosphamide) in the other 14 cases. Immune reconstitution of quadruple group and triple group at 1,3, 6, 9,12 month after transplantation were examined by flow cytometer, and the diference of the two group were estimated with Independent-Samples T test. The incidence of GVHD of the two group was esitimated with Mann-Whitney Test. Kaplan-Meier survival analysis model was used to estimate the overall survival and the disease-free survival (DFS).
Results Immune reconstitution after transplantation of quadruple group and triple group have no significant difference (P&gt;0.05). Acute GVHD (aGVHD) occurred in 9 patients (56.25%) of the quadruple group and in 11 (73.33%) of the triple group, respectively. The incidence of acute GVHD (aGVHD) differed little between the two group (P=0.238). The incidence ofIII~IV°aGVHD in the two group were 6.30% and 26.67%, respectively, and there was no significant difference (P=0.122). 6 patients had chronic GVHD (cGVHD), in the16 cases who could be followed up in quadruple group, 3 of the 11 patients who could be followed up in triple group developed cGVHD postoperatively (P=0.580). Four patients of quadruple group died of hemorrhagic cystitis, mycotic pneumonia, tuberculosis and relapse, respectively. 3 patients of triple group died of GVHD, and the other 3 died of GVHD associated interstitial pneumonia, cytomegalovirus (CMV) pneumonia and pneumocystis carinii infection. The lethality of GVHD of quadruple group and triple group were 0%,26.7%, respectively, and there was significant difference(P=0.027). The one-year disease-free survival rate was 75% and 60% in patients of the quadruple and the triple group, respectively, and significant difference was not noted (P= P=0.188).
Conclusion Compared with triple therapy with CsA, MTX and ATG, CsA+MTX+MMF+ATG procedure dose not worsen the immune reconsititution after transplantation. It can’t decrease the incidence and severity of aGVHD, but can lower the lethality of GVHD in URD-HSCT. The quadruple procedure may lead to higher relapse rate after URD-HSCT.
The Receptor for Hyaluronic Acid Mediated Motility (RHAMM/CD168) Is a Potential Target for Immunotherapy of Patients with B-Cell Chronic Lymphocytic Leukemia.
Authors: Giannopoulos Krzysztof , Hus Iwona , Li Li , Bojarska-Junak Agnieszka , Greiner Jochen , Rolinski Jacek , Dmoszynska Anna , Doehner Hartmut , Schmitt Michael 
Published: April 2020

Abstract
Definition of appropriate target antigens might open new avenues to antigen targeted immunotherapies for patients with B-cell chronic lymphocytic leukemia (B-CLL). We screened the mRNA expression of tumor associated antigens (TAAs), from the literature (fibromodulin, survivin, OFA-iLRP, BAGE, G250, MAGE1, PRAME, proteinase, Syntaxin, hTERT, WT-1), and TAAs defined earlier by serological analysis of cDNA expression libraries from leukemic cells (PINCH, HSJ2, MAZ, MPP11, RHAMM/CD168, NY-Ren60). Peripheral blood mononuclear cells from 43 B-CLL patients and 20 healthy volunteers (HVs) were examined by conventional and quantitative RT-PCR. mRNA of RHAMM/CD168, fibromodulin, syntaxin and NY-Ren60 was expressed in 55–90%, mRNA of HSJ2, MAZ and OFA-iLRP in 90–100% of the patients. No expression of WT-1, h-TERT, BAGE, G250, MAGE1 and survivin was observed. Low (2–20%) expression frequencies of MPP11, PINCH, PRAME and proteinase were detected. RHAMM/CD168, fibromodulin, PRAME and MPP11 showed expression in B-CLL patients, but not in HVs. Because of the exquisite tissue expression of RHAMM/CD168 and its high expression frequency in B-CLL patients even in early stages of disease, mixed lymphocyte peptide culture (MLPC), enzyme-linked immunosorbent spot (ELISPOT) and flow cytometry were performed for antigen specific T cells. In MLPC, RHAMM specific responses by CD8+HLA-A2/R3tetramer+CCR7-CD45RAhigh effector T cells were detected. Moreover, we observed an enhanced RHAMM/CD168 specific CD8+ T cell response after vaccination in one from four HLA-A2 positive B-CLL patients vaccinated with tumor cell lysate pulsed dendritic cells. Therefore, RHAMM/CD168 is an interesting candidate antigen for future immunotherapies in both ZAP-70 (+) and ZAP-70 (−) B-CLL patients.