bluuebunny/crossref_metadata_2025_split huggingface dataset:
Chemotherapy Does Not Enhance CD34+ Cell Collection When Added to Growth Factor and Plerixafor in Patients Who Are Poor Mobilizers
Authors: Rosenbaum Eric R , Nakagawa Mayumi , Pesek Gina , Theus John W , Barlogie Bart , Cottler-Fox Michele H. 
Published: October 2019

Abstract
Abstract 4388

Background:
Many patients who are poor CD34+ cell mobilizers receive the CXCR4 inhibitor plerixafor to enhance mobilization. Here we examined whether chemotherapy followed by growth factor (GF) + plerixafor for mobilization improves CD34+ cell collection in these patients.

Materials and Methods:
We reviewed CD34+ cell collection data from January 2009 through June 2011 for all patients (n=152) at our institution who received plerixafor with GF ± chemotherapy. The primary disease in both groups was multiple myeloma, with 14 non-myeloma patients in the chemotherapy group and 8 non-myeloma patients in the no-chemotherapy group. There were 82 collections preceded by mobilization with chemotherapy (53 male, 29 female), and 80 collections without chemotherapy (51 male, 29 female). Several patients collected on more than one occasion. Age range in the chemotherapy group was 36–76y (median 64) and in the no-chemotherapy group was 42–83y (median 63). Our standard protocol is to collect with plerixafor for 4 consecutive days using a large volume leukapheresis (30L processed). Of the total 162 collections, CD34+ cells collected on each of the 4 consecutive days were recorded. If during the collection process a day was skipped, if less than 4 days of collection were performed, or a collection event on a particular day processed any blood volume other than 30L, all days including and subsequent to such events were excluded from analyses. CD34+ cells were quantified by flow cytometry. Mean CD34+ cells collected on days 1–4 were recorded separately for both groups and compared using two-tailed student’s t-tests.

Results:
Mean number of CD34+ cells collected for the chemotherapy group on days 1–4 were 2.65 (n=82), 2.27 (n=69), 1.60 (n=47), and 1.25 × 106 CD34+ cells/kg (n=38), respectively. Mean number of CD34+ cells collected for the no-chemotherapy group on days 1–4 were 2.51 (n=80), 1.83 (n=67), 1.22 (n=45), and 0.97 × 106 CD34+ cells/kg (n=28), respectively. There were no statistically significant differences between mean CD34+ cells collected on days 1–4 between the two groups (all p-values >0.05). We also asked whether the decrease in mean CD34+ cell yield observed as collection progressed was statistically significant. This phenomenon was determined significant (by one-way ANOVA) for both the chemotherapy and no-chemotherapy groups tested separately (p-values 0.02 and 0.003, respectively).

Conclusions:
Chemotherapy plus GF and plerixafor does not improve CD34+ cell collection in poor mobilizers compared to GF+plerixafor alone. Additionally, mean number of CD34+ cells collected decreases each day, regardless of which of the two mobilization regimens is used.

Disclosures:
Pesek: Genzyme: Membership on an entity’s Board of Directors or advisory committees. Cottler-Fox:Genzyme: Membership on an entity’s Board of Directors or advisory committees.
Recovery of Viable MSCs Isolated From Fresh Umbilical Cord Tissue, Measured After Cryopreservation, Is on Average 8-Fold Higher when Compared to Recovery of Viable MSCs Isolated From Previously Cryopreserved Umbilical Cord Tissue
Authors: Briddell Robert , Litkenhaus Frank , Foertsch Gail , Fuhrmann Angela , Foster Karen , Falcon Girard Kate , Fiscus Bruce , Boehm Ann , Brown Margaret , Pettit Mystie , Rigas Bridges Asimena , Nichols Karen , Fodor William , Kraus Morey 
Published: October 2019

Abstract
Abstract 4398

Background.
The human umbilical cord (hUC) is comprised of a gelatinous connective tissue that contains mesenchymal-like stem cells (TC-MSCs), proteoglycans and collagen. Isolation and cryopreservation of TC-MSCs for potential therapeutic use is an active area of research in the stem cell banking industry. We have compared two methods of cryopreservation to determine the optimal method for storing TC-MSCs: (1) cryopreservation of isolated TC-MSCs from fresh hUC tissue; and, (2) cryopreservation of intact hUC tissue followed by TC-MSC isolation.

Methods.
hUCs were aseptically obtained, and parallel hUC segments from the same hUC were processed by one of the two methods. Method 1: TC-MSCs were isolated from partially dissected fresh hUC segments by overnight collagenese digestion (2.5mg/mL-g), filtration and centrifugation. Isolated TC-MSCs from this process were cryopreserved in a cryopreservation solution (11% DMSO, 1% Dextran40, 4% HSA in 0.9% NaCl), frozen and stored at −80°C for < 30 days. Cells were rapidly thawed at 37°C, washed and resuspended in D-PBS for flow cytometric analysis. Method 2: hUC tissue was cryopreserved in the cryopreservation solution as in Method 1, frozen and stored at −80°C for < 30 days. Frozen hUC tissue was rapidly thawed at 37°C and processed as described above to isolate the TC-MSCs. TC-MSCs recovered from each procedure were enumerated and phenotyped by flow cytometry.

Results.
The recovery of viable CD45−/CD90+ TC-MSCs (after thawing) from fresh hUC tissue (Method 1), on average resulted in 5.04×105 cells/gram of tissue (n=15), whereas the yield of CD45−/CD90+ TC-MSCs from cryopreserved/thawed hUC tissue (Method 2) resulted in 8.20×104 cells/gram of tissue (n=15). Cell viabilities from both processes were comparable.

Conclusions.
Significantly higher (p<0.001) viable cell recoveries (8.4-fold average, 6.8-fold median, range 2.1–18.1) were obtained when TC-MSCs were harvested prior to cryopreservation.

Disclosures:
Briddell: PerkinElmer Inc.: Employment. Litkenhaus:PerkinElmer Inc.: Employment. Foertsch:PerkinElmer Inc.: Employment. Fuhrmann:PerkinElmer Inc.: Employment. Foster:PerkinElmer Inc.: Employment. Falcon Girard:PerkinElmer Inc.: Employment. Fiscus:PerkinElmer Inc.: Employment. Boehm:PerkinElmer Inc.: Employment. Brown:PerkinElmer Inc.: Employment. Pettit:PerkinElmer Inc.: Employment. Rigas Bridges:PerkinElmer Inc.: Employment. Nichols:PerkinElmer Inc.: Employment. Fodor:PerkinElmer Inc.: Consultancy. Kraus:PerkinElmer Inc.: Employment.
Molecular Analysis of the t(2;8)/MYC-IGK Translocation in High-Grade Lymphoma/Leukemia by Long-Distance Inverse PCR
Authors: Kroenlein Hannes , Schwartz Stefan , Reinhardt Richard , Molkentin Mara , Goekbuget Nicola , Hoelzer Dieter , Thiel Eckhard , Burmeister Thomas 
Published: October 2019

Abstract
Abstract 4407

Background:
Burkitt lymphoma and a subset of diffuse large B-cell lymphomas are characterized by chromosomal alterations affecting the MYC oncogene on 8q24. In most cases MYC is found juxtaposed to the immunoglobulin heavy chain (IGH) gene locus. Translocations to the immunoglobulin kappa (IGK) gene locus on 2p11 are observed in around 10% of cases. Little data exist on the molecular mechanisms leading to this aberration. The chromosomal breakpoints on chromosome 8 have been found dispersed over a large area 3’ of MYC. Currently, molecular cytogenetics (FISH) and cytogenetics are the methods of choice to detect the t(2;8) translocation and until now there exists no PCR method to reliably detect it.

Objectives:
In order to obtain a better understanding of this chromosomal translocation we developed a long-distance inverse (LDI) PCR method for the identification of chromosomal translocations affecting the IGK locus. The LDI PCR method takes advantage of the fact that the chromosomal breaks occur on chromosome 2 in the vicinity of the IGK joining segments. Using this method we investigated a number of cytogenetically mostly uncharacterized high-grade lymphoma samples.

Results:
A MYC-IGK juxtaposition was identified in 7 patients and three t(2;8)-positive cell lines. The chromosomal breakpoints were molecularly characterized and analyzed. The linear distance of the breakpoints on chromosome 8 to MYC ranged from some 100 bp to more than 0.5 MB. The breakpoints appeared to be clustered in distinctive regions, however, the number is still to small to draw definitive conclusions. The reciprocal translocated allele could be characterized in the majority of cases and putative break mechanisms are proposed. No larger sequence homologies were detected at the break sites. A minority of breaks were located near putative recombination signal sequences (RSS).

Conclusions:
This study represents the largest series of t(2;8)-positive cases analyzed so far and the LDI PCR developed therein is the first universally applicable PCR-based method to detect this aberration. This LDI PCR should furthermore in general be useful for the molecular analysis of chromosomal translocations affecting the IGK locus. These aberrations have been detected in various lymphoma entities but are currently largely unexplored.

Disclosures:
No relevant conflicts of interest to declare.
Molecular Characterization of Novel Chromosomal Translocations Involved with TCR Locus in T-ALL
Authors: Zheng Haitao , Chen Shaohua , Yang Lijian , Li Bo , Przybylski Grzegorz K , Schmidt Christian A , Li Yangqiu 
Published: October 2019

Abstract
Abstract 4409
Chromosomal translocations occur frequently in hematological malignancies, which might involve in cell transformation and initiation of malignancy. In T-cell malignancies, the chromosomal aberrations are more complex due to the TCR gene rearrangements during T-cell differentiation, which provide different breakpoints for the gene recombination. The chromosomal translocations that occur in the cases of T-cell acute lymphocatic leukemia (T-ALL) frequently involve the juxtaposition of enhancer elements and strong promoter from T-cell receptor (TCR) genes on chromosome 14 (TCRA and TCRD) and chromosome 7 (TCRB and TCRG) with transcription factors genes. Using a novel rapid approach of fine-tiling array comparative genomic hybridization (fine-tiling aCGH) with the array coving the TCR&brkbar;Á&brkbar;Ä, TCR&brkbar;Ã and TCR&brkbar;Â loci, chromosomal alterations were analyzed and the breakpoints and their position in the chromosomes in genomic DNA were identified from patients with T-ALL.
Combining with ligation-mediated PCR (LM-PCR) and sequencing techniques, we clarified at the molecular level novel chromosomal translocation t(X;14)(q11.2;q11), inv(14)(q11;q11), t(1;14)(p32;q11), inv(14)(q11;q32) and t(7;14)(q34;q32) in 4 cases with T-ALL. which caused a V&brkbar;Á37 segment fused to ZFRP1 gene locus, J&brkbar;Á61 segment fused to RPSAP4 gene locus, V&brkbar;Â20 segment rearranged to Ig&brkbar;Ê gene segment and so on. The biological function of the novel fusion genes is needed further research.

Disclosures:
Zheng: the Fundamental Research Funds for the Central Universities (No. 21610603): Research Funding; National Natural Science Foundation of China (no. 30871091): Research Funding. Li:National Natural Science Foundation of China (no. 30871091): Research Funding; Fundamental Research Funds for the Central Universities (No. 21610603): Research Funding.
IL10-592 CC Genotype In the Donor Is Associated with a Significant Decrease of Relapse Rate and a Significant Increase of Event-Free Survival and Overall Survival After Allogeneic Hematopoietic Stem Cell Transplantation In Children with Hematological Malignancies
Authors: Gruhn Bernd , Voigt Janine , Pfaffendorf-Regler Nadine , Wolff Ilona , Zintl Felix , Beck James F. 
Published: October 2019

Abstract
Abstract 4459
IL10 is a pivotal immunomodulatory cytokine and is usually regarded as a suppressor of the immune responses. However, IL10 has been shown to have some immunostimulatory effects. The IL10-592 CC genotype is associated wit higher production of IL10. Because IL10 may promote the development of alloimmunity we hypothesized that the IL10-592 CC genotype in the donor reduces the risk of relapse after hematopoietic stem cell transplantation (HSCT). A cohort of 211 children (median age, 12 years) with acute lymphoblastic leukemia (n=100), acute myeloid leukemia (n=62), myelodysplastic syndrome (n=30) or chronic myeloid leukemia (n=19) who underwent allogeneic bone marrow (n=153) or peripheral blood stem cell transplantation (n=58; T-cell depleted: n=26) in a single center and/or their respective donors was genotyped of IL10 gene for rs1800872 using TaqMan real-time polymerase chain reaction. The donor was HLA-matched unrelated in 48% of transplants and HLA-identical related in 42% of transplants. Conditioning regimen was myeloablative in all cases. Two forms of post-transplant immunosuppression predominated, cyclosporine A and methotrexate in 69% of transplants and cyclosporine A alone in 17% of transplants. Cell samples from the donor were available in 174 cases and from the patient in 197 cases. The IL10-592 CC genotype was present in 82 of the 174 donors (47.1%) and in 104 of the 197 patients (52.8%). Interestingly, we found a significantly reduced incidence of relapse in patients who were transplanted from a donor with the IL10-592 CC genotype (15.9% versus 30.4%; p=0.016). In addition, we observed a significant increase of event-free survival (52.4% versus 33.7%; p=0.019) and a significant increase of overall survival (54.9% versus 37.0%; p=0.040) if the IL10-592 CC genotype was present in the donor. The occurrence of the IL10-592 CC genotype, in either donors or recipients, had no significant impact on treatment related mortality, acute and chronic graft-versus host disease. In conclusion, IL10-592 CC genotype in the donor is associated with a significant decrease of relapse rate and a significant increase of event-free survival and overall survival after HSCT in children with hematological malignancies. This is the first study to describe an association of IL10 gene polymorphism with relapse rate after HSCT. Selecting a donor with the IL10-592 CC genotype could be a useful therapeutic strategy for improving the final outcome after allogeneic HSCT.

Disclosures:
No relevant conflicts of interest to declare.
Impact of KIR B/X Genotype and Missing KIR Ligands in HLA-Matched Unrelated Donor HSCT
Authors: Li Weiyang , He Jun , Bao Xiaojing , Zhou Lina , Depei Wu , Sun Aining 
Published: October 2019

Abstract
Abstract 4488

Objective
To study the prognostic impact of missing ligands for inhibitory killer immunoglobulin-like receptor(KIR) and donor KIR B/X genotype in HLA-matched hematopoietic stem cell transplantation(HSCT) using unrelated donor.

Methods
HLA genotype of 51 patients (ALL 22 cases,AML 13 cases,CML 14 cases,MDS 1 case and HAL 1 case) and their matched unrelated donors was determined by polymerase chain reaction sequence oligonucleotide probes(PCR- SSOP) and sequence specific primers (PCR-SSP).The KIR genotype was determined by PCR-SSP.

Results
Patients were divided into two groups. Group A consisted of 16 patients with donor KIR B/X genotype and missing HLA class I ligands for donor inhibitory KIR, and group B consisted of 35 patients without KIR B/X or missing KIR ligands.The 3-year overall survival(OS) rate for group A was 93.8%, and the OS rate for group B was 61.7% (P=0.058).The 3-year continuous complete remission(CCR) rate for the two groups was 83.3% and 57.2%,respectively(P=0.098).There was no significant difference in neutrophil and platelet recovery,≥ III° acute GVHD(6.3% vs 20.0%,P>0.05) and extensive chronic GVHD (25.0% vs 17.1%,P>0.05) between the two groups.

Conclusions
In HLA-matched unrelated donor HSCT,donor KIR B/X genotype and missing KIR ligands may be associated with decreased severely acute GVHD,improved CCR and OS.

Disclosures:
No relevant conflicts of interest to declare.
CD56-Positive Acute Myeloid Leukemia Has Inferior Outcome After Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Tan Tran-Der , Wu Mau-Ching , Chiou Lun-Wei 
Published: October 2019

Abstract
Abstract 4493

Background:
CD56-positive AML patients have more extramedullary infiltrate and inferior outcomes when treated with intensive chemotherapy. We evaluate the outcomes between CD56 positive and negative AML patients after allogeneic hematopoietic stem cell transplantation.

Methods:
Forty-one AML patients underwent hematopoietic stem cell transplantation at our institute between August 2002 and September 2010. Two of them received autologous transplantation and the other 39 were allotransplant. We analyze the survival and relapse rates when transplanted in complete remission between CD56 positive and negative patients.

Results:
Among these 41 patients, 13 were CD56 positive (M/F 7/6 and median age 34.9 years) with 7 of them (53.8%) had extramedullary involvement; 28 were CD56 negative (M/F 14/14) and median age 42.7 years) with 2 of them (7.1%) had extramedullary involvement. Two-year and five-year overall survivals for all patients were 48% and 40%, respectively. For CD56 positive and negative patients, 2-year overall survivals were 22% and 54%, respectively. There were 30 patients undergoing transplant in complete remission with 8 CD56 positive and 22 CD56 negative, and the relapse rates after transplant were 75.0% and 31.8%, respectively, (p = 0.049). High risk cytogenetics were found in 5 out of 13 CD56 positive patients and 5 out of 17 available cytogenetics CD56 negative patients, which is of no statistically different.

Conclusion:
Even following the most intensive treatment hamatopoietic stem cell transplantation, CD56-positive AML patients still have more relapse rate, and poor survival which is independent of cytogenetic risk. This indicated CD56 positivity is truly a poor prognostic factor and needs to consider allogeneic hematopoietic stem cell transplantation early on remission achieved and earlier immunosuppressants withdrawal is mandatory for those who allotransplant.

Disclosures:
No relevant conflicts of interest to declare.
Outcomes of Hematopoietic Stem Cell Transplantation for Childhood Acute Lymphoblastic Leukemia with t(4;11)
Authors: Sekiya Yuko , Narita Atsushi , Yoshida Nao , Matsumoto Kimikazu , Kato Koji 
Published: October 2019

Abstract
Abstract 4502

Introduction:
As the prognosis of childhood acute lymphoblastic leukemia (ALL) with t(4;11) is generally poor, hematopoietic stem cell transplantation(HSCT) at 1st complete remission is recommended in most of Japanese ALL protocol. We analyzed outcomes of HSCT for childhood ALL with t(4;11) at our department, and assessed the outcomes with age and preconditioning at HSCT.

Subjects:
We retrospectively analyzed 15 patients who underwent HSCT between 1991 and 2011. The onset of ALL was less than one year old (infant ALL) in 9 patients and over one year old in 6 patients. The disease status at HSCT was 1st complete remission (CR1) in 10 patients, 2nd complete remission (CR2) in 2 patients, and more advanced stages in 3 patients. 13 patients underwent allogeneic HSCT and 2 patients underwent autologous HSCT at median age of 19 months. In allogeneic HSCT, the number of bone marrow transplantation from HLA matched sibling was 3 and 4 from unrelated donors, and that of cord blood transplantation was 6. Nine patients including 7 infant ALL underwent conventional stem cell transplantation (CST), 6 patients including one infant ALL underwent reduced intensity stem cell transplantation (RIST). Preconditioning of RIST was mostly consisted of fludarabine and melphalan with 3Gy of total body irradiation (TBI).

Results:
Of all 15 patients, 8 patients are alive in CR. Three out of 9 infant ALL and 5 out of 6 ALL patients, whose onset was above 1 year old, are alive in CR. Seven of 10 ALL patients who underwent HSCT in 1st CR are alive, on the other hand, only one of 5 patients in 2nd CR or non CR is alive. Five patients who underwent RIST over 1 year old are alive in CR.

Discussion:
HSCT for childhood ALL with t(4;11) in 1st CR could achieve better outcome than those in 2nd CR or non CR. The transplant outcomes with RIST was better than CST and it could be applied for ALL patients with t(4;11) whose onset is over one year old.

Disclosures:
No relevant conflicts of interest to declare.
KIR Ligation During Ex Vivo Expansion Allows Molding Of The KIR Repertoire
Authors: Lee Dean A. , Senyukov Vladimir V , Trembley Jerome R 
Published: October 2019

Abstract
NK Cell subpopulations express tremendous diversity through polymorphisms, haplotypes, differential expression, and licensing of the Killer Immunoglobulin-like Receptors (KIR). KIR diversity affects both the predisposition to cancer, and the response to therapies such as hematopoietic stem cell transplantation. Clinical trials that take advantage of the anti-cancer properties of NK cells have been limited to choosing donors on the basis of KIR genotypes and/or HLA haplotypes. Moreover, adoptive immunotherapy approaches have been limited by low NK cell doses. The latter hurdle has been recently mitigated by methods for expanding clinical grade NK cells ex vivo. These approaches for growing large numbers of cells now enable investigation into selecting more potent NK cell subsets for increased therapeutic efficacy.
We hypothesized that the desired KIR repertoire could be molded through inhibition of undesirable KIR populations by crosslinking with relevant anti-KIR antibodies during expansion with our previously described method, which produces a mean 30,000-fold expansion of NK cells in 3 weeks.
First, we determined that maximum inhibition was obtained when anti-KIR antibodies were applied to previously activated NK cells, crosslinked with secondary antibody, and then restimulated for proliferation.
Robust reduction of targeted KIR-positive populations could be achieved for each inhibitory KIR (Fig. 1). When pre-activated with anti-KIR2DL1 for one stimulation cycle, NK cells expressing this KIR were decreased by a median of 70.4% ± 19.3%. Similarly, KIR2DL2/3+ NK cells could be reduced by 56% ± 17.5%, and KIR3DL1+ NK cells could be reduced by 53.5% ± 16.3%. When anti-KIR antibodies were combined, similar suppression of multiple-KIR subpopulations was observed. Other NK cell receptors were not significantly affected during targeted KIR inhibition.
We then assessed the resulting NK cell populations for degranulation responses to targets with selected HLA as KIR ligands. Inhibition of KIR-expressing subpopulations during expansion resulted in NK cell populations with enhanced degranulation against tumor cells expressing the HLA ligand of the targeted KIR. Importantly, the cytotoxicity of the bulk NK cell population against HLA-negative targets remained.
These results indicate that KIR crosslinking during NK cell propagation enables significant reduction in the targeted KIR subpopulations, resulting in an NK cell population with a selective decrease in KIR inhibition. By utilizing antibody-controlled expansion for molding of the KIR repertoire according to patient HLA type, a personalized NK cell product may be produced with enhanced potency, improving NK cell immunotherapy.

Disclosures:
No relevant conflicts of interest to declare.