bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Arrest of Cell Cycle Progression in Mixed Lymphocyte Reaction by Cultured Mouse Mesenchymal Stem Cell.
Authors: Kim Jeong-A. , Hong Sungyoul 
Published: October 2019

Abstract
Mesenchymal stem cells (MSCs) possess in vitro immunosuppressive properties that appear not to be major histocompatibility complex (MHC)restricted. This study evaluated the immune suppressive effect of mouse MSCs on mixed lymphocyte reaction (MLR), and the mechanisms were investigated. MSCs were obtained from BALB/c bone marrow and cultured in low-glucose DMEM media. The expression of surface antigens and cell cycle were analyzed by flow cytometry. The MSC-induced suppression was assessed by MLR and transwell culture. The BALB/c MSCs constitutively expressed MHC class I and CD54 (ICAM-1) antigens but were negative for MHC class II, CD40, CD80 (B7–1) and CD86 (B7–2) antigens. MSCs suppressed allogeneic C57BL/6 T lymphocytes proliferation by adding them to MLR in which C3H spleen cells were used as a stimulator. This inhibition was dependent on the dose of BALB/c MSCs but independent of MHC. C57BL/6 T lymphocytes proliferation was still inhibited when BALB/c MSCs were added in culture 3 days after starting of MLR. When MSCs were separated from C57BL/6 T cells by using the transwell membrane, the suppression of immune response wasn’t observed, which suggested that the suppressive effect was dependent on cell-cell contact between BALB/c MSCs and C57BL/6 T cells. When C57BL/6 T lymphocytes were cultured with MSCs, the percentage of C57BL/6 T cells in G0 phase increased from 51.8±7.66% to 77.2±7.39% compared with the case that only C57BL/6 T cells were cultured. When the C57BL/6 T cells were cultured with C3H spleen cells, most of C57BL/6 T cells were in G2/M (96.38±3.33%). But by the addition of MSC to MLR, the percentage of T cells in G2/M decreased to 33.0±9.66% while that of T cells in G0 increased from 2.0±0.71% to 66.2±7.46%. We concluded that the cell cycle of responder T lymphocytes in MLR is arrested at G0 phase by MSCs.
Figure Figure
Glucocerebroside Alleviates Chronic Graft-Versus-Host-Disease in a Murine Model.
Authors: Nagler Arnon , Margalit Maya , Ohana Meir , Alper Ruslana , Pappo Orit , Rabbani Elazar , Engelhart Dean , Ilan Yaron 
Published: October 2019

Abstract
Background: Chronic graft-versus-host-disease (cGVHD) is a major obstacle to successful bone marrow transplantation. NKT lymphocytes may have a role in regulation of the immune attack in cGVHD. Glucocerebroside (GC) is a naturally occurring glycolipid that plays a role in the immune modulation of NKT lymphocytes.
Objectives: To determine the immune modulatory effect of GC in a murine model of cGVHD.
Methods: cGVHD was generated by infusion of 2x107 splenocytes from B10.D2 donor mice into Balb/c recipient mice, which received 7Gy 60Co total body irradiation (TBI) prior to transplantation. Recipient mice were followed for 55 days. Histological parameters of cGVHD-associated cutaneous, liver and bowel injury were assessed. To determine the mechanism of GC-mediated immune modulation, intrahepatic and intrasplenic lymphocytes were isolated and analyzed by FACS for CD4+, CD8+,CD56+ and NKT subpopulations. Serum cytokine levels were determined.
Results: Treatment with GC significantly alleviated cGVHD. GC-treated mice gained weight and manifested a significant decrease in skin cGVHD clinical score (0.91 vs. 1.66 in GC treated vs. control mice, respectively). The beneficial effect of GC was associated with a 58% and a 31% reduction of the intrasplenic and the intrahepatic NKT cell number, respectively, accompanied by a 27% decrease of intrasplenic CD3+CD4+ T cells. Intrasplenic (but not intrahepatic) decrease in NK cell number was also observed in GC treated animals (48% vs. 7%). Administration of GC led to increased serum IFNγ and TNFa levels in GC treated compared to untreated controls (46 vs. 27, and 92 vs. 52 pg/ml, respectively), while IL10 levels decreased (61 vs. 83 pg/ml, respectively).
Conclusions: Administration of GC led to significant amelioration of chronic GVHD. This effect was associated with an altered NKT lymphocyte distribution, decreased numbers of effector T lymphocytes and a shift towards a Th1-type cytokine profile. Glucocerebroside may serve as a new therapeutic measure for amelioration of cGVHD.
Role of IL-4-Producing CD8+ T Cells in Patients with Chronic Graft-Versus-Host Disease (cGVHD) and in Normal Healthy Volunteer.
Authors: Nakamura Kengo , Takebayashi Masashi , Tajima Kenichirou , Tanijiri Tsutomu , Yokoi Takashi , Son Yonsu , Matsumoto Noriaki , Zen Katsuhiro , Kishimoto Yuji , Amakawa Ryuichi , Fukuhara Shirou 
Published: October 2019

Abstract
The pathophysiology of cGVHD still remains unclear. To gain more insight into the immunological mechanism of cGVHD, we examined surface marker and cytokine production of peripheral blood T cells from 19 patients in the chronic phase of allogeneic hematopoietic stem cell transplantation (allo-HSCT). The median age of patients was 31 years (range 13–47 years). The median days from allo-HSCT to examination was 43 months (range 17–94 months). Ten patients in whom 6 have treated with immunosuppressive agents and 4 without them developed chronic GVHD, whereas 9 patients in whom 5 have treated with immunosuppressive agents and 4 without them did not develop chronic GVHD. Peripheral blood mononuclear cells were extracted from patients and 10 healthy volunteers, followed by isolation into CD4+ T cells and CD8+ T cells. The analysis of surface marker and intracellular cytokine production of these cells was performed using fluorescence activated cell sorter. The percentage of IFN-g-producing CD8+ T cells among CD8+ T cells was significantly higher in patients with or without cGVHD than in healthy volunteers (p<0.001). Since all 19 patients were in remission, the IFN-g-producing CD8+ T cells might be related to graft-versus-leukemia effect. On the other hand, the percentage of IL-4-producing CD8+ T cells among CD8+ T cells was significantly higher in patients with cGVHD (mean 3.3%, range 1.3–8.2%) than in patients without cGVHD (mean 1.2%, range 0.8–1.7%) and healthy volunteers (mean 1.1%, range 0.1–1.6%) (both p<0.001). By contrast, the percentage of IL-4-producing CD4+ T cells was not different among patients with and without cGVHD and healthy volunteers. Then, we minutely explored the immunological role of IL-4-producing CD8+ T cells in healthy volunteers. These cells expressed the surface marker of CD25, CCR4 and CTLA-4. In the analysis of cytokine production in these cells, we identified the type 2 cytokine, such as IL-4, IL-5, IL-10, and IL-13, although we could not find IFN-g. In conclusion, these findings suggest that IL-4-producing CD8+ T cells may be an immunological marker of cGVHD in which these cells may play a crucial role as regulatory T cells.
Protective Effect of Curcumin in an F1-Hybrid Model of Acute Graft-vs.-Host Disease.
Authors: Ellison Cindy A. , Natuik Shannon A. , Fischer Jacqie M. , Gartner John G. 
Published: October 2019

Abstract
Curcumin (diferuloylmethane), a pigment from the rhizomes of Curcuma Longa, has anti-oxidant, anti-tumor and anti-inflammatory properties. It has been shown that curcumin can inhibit the development of experimental allergic encephalomyelitis, a Th1-mediated, autoimmune, demyelinating disease that affects the central nervous system in mice. Because acute GVHD involves the development of a Th1-mediated immune response, and is characterized by the development of a potent, systemic, inflammatory response, we wished to determine whether curcumin might also protect mice from developing acute GVHD in the C57BL/6→(C57BL/6 x DBA/2)F1-hybrid model. We found that ip injections of curcumin (100 micrograms dissolved in 25 microliters of DMSO), given once a day, every 2 days, protected recipients from developing acute, lethal GVHD. Two thirds of the treated group survived well past day 100 post-transplantation. Control GVH mice treated with DMSO alone became moribund within three weeks of transplantation, the time at which mice with acute GVHD typically succumb. To ensure that the protective effects of curcumin were not due to abortion of the graft, we analyzed T cell engraftment in some of the long-term survivors. In the surviving mice tested on days 135 and 275 post-transplantation, we were unable to detect any cells of recipient origin in the spleen. Furthermore, the percentage of donor-derived CD4+ cells ranged from 32–47% and the percentage of CD8+ T cells ranged from 25–42% in the non-adherent spleen cell fraction. Data from these experiments suggest that curcumin can protect mice from developing acute, lethal GVHD.
Genetic Susceptibility Loci for Lethal Graft-Versus-Host Disease in MHC-Identical Mice: Initial Results from a Genome-Wide Scan.
Authors: Cao Thai M. , Pang Wendy W. , Grumet F. Carl , Lazzeroni Laura C. , Shizuru Judith A. 
Published: October 2019

Abstract
Minor histocompatibility antigens (mHAg) are T-cell peptide antigens derived from polymorphic endogenous gene products encoded throughout the genome. Despite extensive human genetic diversity, recent experimental evidence suggests that a mHAg set may be limited in size because alloimmune responses directed against them can be dominated by only a few specific peptides. To extend upon these observations, we performed a genome-wide scan to identify susceptibility loci for graft-versus-host disease (GVHD) in a murine model. We previously identified B10.BR (H2k) mice to be resistant, and BALB.K (H2k) mice susceptible, to lethal GVHD when transplanted with hematopoietic cells from MHC-matched AKR/J (H2k) donors (Cao et al. PNAS2003:11571). In the present study B10.BR, BALB.K, or (B10.BR x BALB.K)F1 mice were lethally irradiated and injected with AKR/J bone marrow at radioprotective doses plus 2.0 or 3.0 x 106 purified splenic T-cells. Confirming our prior observations, B10.BR mice survived to day 60 whereas all BALB.K mice died prior to day 10 from GVHD. All (B10.BR x BALB.K)F1 mice also died after a median of 10 days, indicating that GVHD susceptibility in BALB.K mice is a dominant trait. (B10.BR x BALB.K)F1 mice were then backcrossed to the GVHD-resistant B10.BR parent. Resulting [B10.BR x (B10.BR x BALB.K)]BC mice (n = 180) were genotyped and similarly irradiated and injected with AKR/J bone marrow plus 3.0 x 106 splenic T-cells. Forty (22%) BC mice died with signs of GVHD prior to day 100. Surprisingly, a 90-marker genome-wide scan for this phenotype, lethal GVHD prior to day 100, revealed highly significant linkage (P <.001) only to chromosome 16. Linkage analysis was performed with MapManager QTX 0.29 using Kosambi’s map function. As shown in Figure 1, interval mapping for chromosome 16 with additional markers revealed a broad and complex linkage pattern encompassing 2 markers, D16Mit4 (27.3 cM) and D16Mit189 (55.2 cM), that had peak LOD scores of 8.8 and 8.0 respectively. Another genetic locus on chromosome 13 flanked by markers D13Mit248 and D13Mit149 with a peak LOD score of 2.3 by interval mapping was suggestively linked. While an evaluation of candidate genes within the support intervals is in progress, these results thus far suggest that the critical gene dose required for inducing lethal GVHD may be remarkably small. If validated as encoding for immunodominant mHAg, these results furthermore suggest that a functionally relevant mHAg set may be definable for clinical genotyping.
Reproducibility and Robustness of the Xcellerate III Process for the GMP Manufacture of Xcellerated T Cells™ for Infusion into CLL Patients.
Authors: Hami Lisa , Green Cherie , Miller Katharine , Craig Stewart 
Published: October 2019

Abstract
Autologous peripheral blood mononuclear cells (PBMC) cryopreserved from a leukapheresis collection comprise the starting cellular source for the Wave® Bioreactor-based Xcellerate III Process [Hami et al, Bioprocessing Journal2003: 2; 23–32] used for the GMP manufacture of Xcellerated T Cells. For an ongoing clinical trial, n=13 patients have been infused with products manufactured from PBMC cryopreserved and stored in the vapor phase of liquid nitrogen (LN2) for 2–9 days before use in the Xcellerate III Process. This clinical protocol was recently amended to allow patients to receive a 2nd infusion of Xcellerated T Cells. To date, 2nd products have been manufactured for 11 of the CLL patients using the original PBMC that had been stored cryopreserved for up to 7 months from collection. Comparison of the processes for the manufacture of 1st (n=13) and 2nd (n=11) infusion products shows:
• No significant difference in the in-process T cell activation as determined by increase in cell size, up-regulation of CD25 & up-regulation of CD154 expression (refer to Figure 1).
Figure Figure
• The total cell yield for 2nd infusion products is within one (1) standard deviation of the average for the manufacture of the 1st infusion product (refer to Table 1).
• No significant difference in cell viability, CD3+ purity, or CD4:CD8 ratio for the final Xcellerated T Cells product (refer to Table 1).
Table 1. Final Product Characteristics
Final Product (Day 13) Average±S.D. p value 1st Infusion (n=13) 2nd Infusion (n=11) Total Cell Yield (x109) 0.01 137 ± 35 104 ± 17 Cell Viability (%) 0.15 93.5 ± 3.4 91.6 ± 2.4 CD3+ Purity (%) <0.001 98.4 ± 1.1 99.0 ± 0 CD4:CD8 Ratio 0.32 8.5 ± 7.7 5.7 ± 4.5
These data demonstrate high reproducibility and robustness of the Xcellerate III Process when using PBMC from the same leukapheresis collection in sequential processing runs. In addition, these data demonstrate that cryopreserved PBMC can be stored for many months prior to their use as the starting material in the Xcellerate III Process. Xcyte™, Xcyte Therapies™, Xcellerate™, Xcellerated T Cells™ and the circle logo are trademarks of Xcyte Therapies, Inc.
Mobilization of Peripheral Blood Progenitor Cells (PBPC) with G-CSF and GM-CSF in Autologous and Allogeneic Pediatric Transplants.
Authors: Echevarria Maria E. , Gonzalez Javier , Barrios Nilka J. 
Published: October 2019

Abstract
BACKGROUND: Chemotherapy and/or use of single or multiple growth factors have simplified the mobilization and collection of PBPC for autologous and allogeneic transplantation following high-dose chemotherapy. Nevertheless, the best method for PBPC mobilization continues uncertain. In children, mobilizations for CD34+ cells for autologous or allogeneic PBPC has been almost exclusively done with G-CSF. We conducted a phase III study to evaluate the results achieved using the combination of G-CSF + GM-CSF mobilizing CD34+ cells in children undergoing autologous or allogeneic PBPC transplantation.
METHOD: A total of 34 pediatric patients participated in the study. Seven healthy donors and 27 pediatric patients with different types of cancer participated in the mobilization study. They included patient with acute lymphoblastic leukemia, acute myelogenous leukemia, advanced stage neuroblastoma, Hodgkin’s lymphoma and rhabdomyosarcoma. Mobilization with G-CSF and GM-CSF (10μg/Kg/day each) was administered for 5 consecutive days in all patients, followed by PBPC harvesting until a threshold of 2.0 x 10^6/Kg CD34+ cells was obtained. A total of 19 PBPC transplants were performed (15 autologous and 5 allogeneic). Neutrophil engraftment was defined as ANC> 500/mm³ and, platelet engraftment was defined as a platelet count > 30,000/mm³ (transfusion independent) for more than 48 hours.
RESULTS: Table 1. The engraftment kinetics for neutrophils and platelets were 9.87 and 12.6 days respectively for the autologous transplants, and 9.4 and 12.2 days for the allogeneic. The mean number of days to achieve a higher or equal to 2 x 10^6 count of CD34+ cells in cancer patients was 2 days and 1 day for healthy donors. Side effects during mobilizations were mild and included nausea or vomiting, and tingling sensations in one individual. No secondary side effects like bone pain, headaches, or flu-like symptoms were experienced or documented. No infections were documented in the transplanted population, both autologous and allogeneic. Acute Graft versus Host Disease (GVHD) was documented in 2 patients that underwent an allogeneic PBPC transplants.
CONCLUSIONS: This study suggest that the combination of G-CSF + GM-CSF (10μ/Kg/day x 5 days) is well tolerated. It seems to be cost effective and adequate to mobilize more than average numbers of PBPC in a shorter period of time (from 1–3 apheresis) to perform allogeneic/autologous hematopoietic transplants in the pediatric population. It furnished a clinical reduction in the degree and duration of severe neutropenia in both allogeneic and autologous transplants as compared with other reports, without added increase incidence of GVHD.
Mobilizations
24 hrs 48 hrs 72 hrs 96 hrs Mean #CD34+ x 10^6/Kg (cancer patients) 3.3 2.56 1.43 0.82 Mean #CD34+ x 10^6/Kg (healthy donors) 6.7 5.1 12.2
Etoposide Containing Preparative Regimen for Allogeneic Hematopoietic Stem Cell Transplantation in Patients with Acute Lymphoblastic Leukemia.
Authors: Saito Takeshi , Asai Osamu , Dobashi Nobuaki , Yano Shingo , Ichiba Tamotsu , Osawa Hiroshi , Takei Yutaka , Sugiyama Katuki , Okawa Yutaka , Takahara Sinobu , Yamaguchi Yuko , Minami Jiro , Usui Noriko , Kobayashi Masayuki 
Published: October 2019

Abstract
[Purpose] Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is one of the powerful therapeutic tools for patients with adult lymphoblastic leukemia (ALL) even though transplant-related toxicity and the recurrence of leukemia are still problematic. In order to improve outcome of patients with adult ALL who received allo-HSCT, we investigated the clinical efficacy and safety of etoposide containing conditioning regimen for allo-HSCT.
[Patients and Methods] Adult patients (including patients with 17, 18 and 19 years of age) with ALL were treated between February 1991 and October 2003 in our institute. When patients attained complete remission (CR) by induction therapies using JALSG ALL protocols or others, they received allo-HSCT [bone marrow (BMT), peripheral blood stem cell (PBSCT), and cord blood transplantation (CBT)]. Conditioning regimen was consisted with etoposide (60 mg/kg), cyclophosphamide (120 mg/kg), and fractionated total body irradiation (10 Gy) (CY/ETP/TBI). As clinically indicated, cytosine arabinoside (CY/ETP/TBI/AraC) or nimustine hydrochloride (CY/ETP/TBI/ACNU) was added to CY/ETP/TBI.
[Results] In the 23 patients [male/female: 15/8, median age was 30 years (range 17–50 years)], 19 patients received BMT (13 from related and 6 from un-related donor), 3 received PBSCT from related donor, and one was performed CBT. As the conditioning regimen, 16 patients received CY/ETP/TBI, 4 received CY/ETP/TBI/AraC, and 3 received CY/ETP/TBI/ACNU. Six out of 23 patients had Philadelphia chromosome positive ALL. Twenty-two patients received HLA serologically identical donor. Remission status at transplant included 16 patients in 1st CR, 5 in 2nd CR, and other 2 patients in non-CR. Grade II toxicity according to Bearman scale was observed in 52% (stomatitis), 22% (gastrointestinal), 9% (bladder), 4% (hepatic), and 4% (cardiac). Critical regimen-related toxicities and fatal veno-occlusive disease were not observed in any patients. The disease-free (DFS) and over-all survival (OS) rate of all patients was 51.1 % and 55.8 %.
[Discussion/Conclusion] These results indicated that an intensified etoposide containing preparative regimen is a feasible regimen and it has good efficacy for transplant recipients with ALL.
Defective Anti-Bacterial Carbohydrate Antibody Production after Unrelated HSC Transplantation.
Authors: Shah Ami J. , Wagner John E. , Kapoor Neena , Chan Raymond , Weinberg Kenneth I. , Parkman Robertson 
Published: October 2019

Abstract
Protective levels of anti-bacterial carbohydrate antibodies are necessary to protect HSC transplantation recipients from severe infections with encapsulated bacteria. Recipients of unrelated HSC transplants are at increased risk of severe infections regardless of their history of graft versus host disease (GVHD). Antibodies to a naturally occurring, prototypic bacterial carbohydrate antigen, polyribose phosphate (PRP), present in Hemophilus influenza type b and K100 stains of E. coli, were measured in the recipients of unrelated bone marrow (UBMT) and unrelated cord blood transplants (UCBT). UBMT and UCBT recipients, who were not on immunosuppressive therapy, were analyzed with follow-ups of 12 and 7 years, respectively. Of 20 UBMT recipients, only 3 had protective anti-PRP antibody levels. All 3 recipients with protective antibody levels were younger than 1 year old at the time of transplantation and did not have a history of GVHD. No recipients, who were older than 1 year old at the time of transplantation, had protective levels of anti-carbohydrate antibody. Of 16 UCBT recipients, 7 had protective antibody levels, including recipients as old as 53 at the time of UCBT. All unrelated recipients, who did not have protective levels of anti-carbohydrate antibodies, were able to produce protective levels of antibodies to a protein antigen, tetanus toxoid, after immunization. Therefore, the inability of the recipients of unrelated HSC to produce protective levels of anti-bacterial carbohydrate antibody is not part of a generalized antibody deficiency state, but is a specific defect. Since bacterial carbohydrate antigens are T lymphocyte-dependent antigens in humans, the results indicate that thymic and T lymphocyte function are greater in UCBT recipients than in UBMT recipients (p=.008 for protective anti-PRP antibody levels in recipients older than age 1). Defects in protective anti-bacterial carbohydrate antibody production are common in the recipients of HSC unrelated transplants regardless of the source of HSC. The recipients of unrelated HSC transplants, especially UBMT, should be tested for their capacity to produce anti-carbohydrate antibody, and, if deficiencies are detected, appropriate prophylactic measures undertaken to reduce the likelihood of severe bacterial infections.