bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Assessment of IL-6R Gene Using FISH in MM.
Authors: Min Hyun Jung , Kim Tae Young , Oh Bora , Lee Jae Hoon , Lee Dong Soon 
Published: October 2019

Abstract
Multiple myeloma (MM) is characterized by proliferation of plasma cell, mostly in the bone marrow. It is established that cytokines including IL-6 play a key role as growth and survival factor for plasma cells through the specific receptors on the cell surface. Abnormalities of chromosome 1q have frequently been reported in various cancers as well as multiple myeloma (MM). Recent studies suggest that amplification of chromosome 1q is a negative prognostic factor. To determine whether increase of the IL-6 receptors located at chromosome 1q21 stimulate secretion of cytokine IL-6 we performed a gene expression evaluation on 136 newly-diagnosed MM, analyzed by interphase FISH for IL-6R(1q21) amplification, Rb1(13q14) deletion, and IGH translocation and additional copies of 9,17,20. We found that the correlation of IL-6R gene and other major genetic risk factor in MM. IL-6R gene amplification (3 to 12 orange signal) was detected in 46 of 136 (34%). These results indicate that IL-6R gene occur the gain of chromosome 1q. It is suggest that the identification of interleukin-6 receptor associated with 1q amplification is useful in providing diagnostic information on multiple myeloma.
Bone Marrow Stem Cell Transplantation Restores Heme Oxygenase and Antioxidants Genes and Pancreatic Function in Type 1 Diabetes.
Authors: Wang Jishi , Peterson Stephen , Kruger Adam , McClung J. , Abraham N.G. 
Published: October 2019

Abstract
Type 1 diabetes is caused by the autoimmune destruction of pancreatic beta(β)cells has been linked to a decrease in circulating stem cells and increased superoxide (O2)secondary to a decrease in heme oxygenase (HO) activity. Decrease HO-1 increases oxidative stress, resulting in β-cell destruction. O2 and β-cell destruction. Nonobese diabetic (NOD) mice (2 months old), which undergo pancreatic T-cell infiltration resulting in elevated were irradiated and reconstituted with bone marrow stem cells from BALB/c mice(<2 months old) using a perfusion technique, exhibited neither insulitis nor overt diabetes. The newly developed T-cells in BMT recipients were tolerant to both donor and host major histocompatability complex determinants. After six months, BMT-treated mice showed the same glucose tolerance as age-matched controls, HO activity was restored to the level of age-matched controls, and EC-SOD protein levels returned to normal. The increased HO activity after BMT was accompanied by a significant elevation of HO-1 protein and a significant decrease in O2-levels(μmol/mg protein 1.16±0.37 μmol/mg protein versus 2.81±0.37 in nontransplant mice). These results demonstrate that bone marrow-derived stem cell transplant restored the antioxidant genes HO-1 and EC-SOD as well as pancreatic function. In addition, we demonstrate that combining BMT and islet cell transplant can restore insulin production in severely diabetic animals. We have thus demonstrated in this animal model of type 1 diabetes that BMT is effective as both a preventive and therapeutic treatment.
Reversal of New-Onset Type 1 Diabetes in Mice and Induction of FoxP3+ Regulatory T Cells by Syngeneic Bone Marrow Transplantation.
Authors: Ouyang Jian , Wen Yanting , Chen Junhao , Liu Yong , Wu De Pei 
Published: October 2019

Abstract
Objective: Autologous bone marrow transplantation (ABMT) was introduced as a novel strategy to treat new-onset type 1 diabetes (T1D). Induction of CD4+CD25+ FoxP3+T regulatory lymphocytes (Tregs) helps maintain self-tolerance in autoimmune diabetes. Here, we were tying to investigate the effects of syngeneic bone marrow transplantation (syn-BMT) in T1D mice models and to investigate the role of Tregs in this context.
Research design and methods: Syn-BMT was used to mimic the ABMT to T1D patients in human. Fraternal inbred multiple low doses of streptozotocin (mld-SZ) induced diabetic mice were used as both donors (n = 7) and recipients (n = 20). And normal fraternal inbred mice were also used as donors (n=13). After total body irritation to recipient mice, syn-BMT was given either on day 10 or on day 40 of T1D. Blood and urine glucose was observed. On day120 after syn-BMT, glucose tolerance was tested and then mice were killed. Insulin levels in sera were tested by ELISA and pancreata histological and morphometry were analyzed by HE staining. Tregs in spleens were tested by Flow cytometry analysis, FoxP3 protein was tested by western blot, and FoxP3 mRNA was tested by real time PCR.
Results: Syn-BMT, if applied when T1D is new-onset (10 d), can reverse blood glucose to close to normoglycemia without farther relapse, maintain blood insulin levels, improve glucose tolerance and ameliorate pancreata destruction. Syn-BMT leading to recovery of T1D results in the induction of Tregs, increased Foxp3 protein and mRNA expression in both the groups of diabetic mice receiving syn-BMT with either diabetic or normal donors. However, if given on a later disease stage (40 d), although syn-BMT helped reduce blood glucose for about 66 d and improved pancreata histological and morphometry, it showed relapse of diabetes further and decrement of insulin levels. Interestingly, this group did not show the increment of Tregs, Foxp3 protein or mRNA expression.
Conclusions: This study provided a rationale to treat human new-onset T1D with autologous HSC. Syn-BMT, if given when T1D is new-onset, is safe and is able to reverse the diabetic status. The induction of Tregs and Foxp3 in both mRNA and protein levels as the results of syn-BMT, may contribute to the immune tolerance after syn-BMT, and the observed improvement of new-onset T1D.
Optimizing Culture Conditions for Ex Vivo Expansion of Virus Specific T Cells.
Authors: Borelli Gabriel , Aarvak Tanja , Brunsvig Anne , Dyrhaug Marianne , Lundby Marianne , Hammerstad Hege , Rasmussen Anne Marie , Kvalheim Gunnar 
Published: October 2019

Abstract
Adoptive immunotherapy with virus specific CD8+ cytotoxic T lymphocytes (CTL) is currently an option for treatment of viral infections after allogeneic stem cell transplantation. A major obstacle for clinical grade production of virus specific CTL is to retain the antigen specific clones, which can be easily deleted during ex vivo expansion. Manipulation of CD3/CD28 engagement, depletion of T regulatory (Treg) cells before expansion and prestimulation with antigen presenting cells were explored in this study. Mononuclear cells were obtained from HLA-A2+ donors by leucapheresis. Lymphocytes were enriched by elutriation, stimulated with anti-CD3/anti-CD28high Dynabeads or Dynabeads® ClinExVivo™ CD3/CD28 and cultured for 10 days in CellGro media supplemented with human AB serum and IL-2. Virus specific CD8+ CTL were quantified by flow using pentamer staining. By using anti-CD3/ anti-CD28high Dynabeads a more balanced expansion of CD4+ and CD8+ subsets was obtained, while Dynabeads ClinExVivo CD3/CD28 allowed a higher expansion of CD8+ subset and therefore a higher expansion of the virus specific CTL. A high Dynabead: T cell ratio (3:1) deleted completely the virus specific CTL. By reducing this ratio we could retain the virus specific CTL after expansion. No benefits were observed by adding extra Dynabeads during expansion. It is known that Treg cells can inhibit expansion of all T cell subsets. By depleting Treg cells with CD25 Dynabeads prior to T cell expansion, we observed a significantly increased expansion of all T cells, including virus specific CTL. In some patients low numbers of virus specific CTL can be detected. To increase the numbers of antigen specific CTL we have included a prestimulation step using peptide-loaded mononuclear cells prior to expansion with Dynabeads, which gave more than 3000- fold expansion of virus specific CTL. We are currently exploring the phenotype profile of expanded CTL (CD62L, CCR7, CD57, CD27, CD28 and CTLA-4), functionality (cytokine secretion and proliferative capacity) and cytotoxicity. This protocol can be upgraded to clinical grade production of virus specific CTL for treatment of viral infections in immunocompromised patients.
InVitro Culture of Human Cord Blood Hematopoietic Stem/Progenitor Cells Interfered with Their Distribution to Other Bone Marrow Compartments after Intra-Bone Marrow Transplantation.
Authors: Ohishi Kohshi , Yamamura Kentaro , Masuya Masahiro , Katayama Naoyuki 
Published: October 2019

Abstract
Intra-bone marrow transplantation (IBMT) is a novel strategy for transplantation of hematopoietic stem cells because it can transfer various types of cells to bone marrow regardless of their homing capacity. However, reconstitution process of these cells after IBMT remains to be fully elucidated. Here, we investigated whether in vitro culture of cord blood hematopoietic stem/progenitor cells affects their reconstitution in bone marrow after IBMT. Freshly isolated AC133+ cells (5x104 cells/mouse) or all cells derived from AC133+ cells cultured with growth factors (stem cell factor, flt-3 ligand, and thrombopoietin) for 5 days were injected into the bone marrow of the left tibia in irradiated NOD/SCID mice. In the bone marrow of the injected left tibia, the engraftment levels of human CD45+ cells at 6 weeks after transplantation was not considerably different between transplantation of noncultured and cytokine-cultured cells (54±28% vs. 69±13%). However, the migration of transplanted cells to the bone marrow of other noninjected bones was extremely lower for cytokine-treated cells compared with noncultured cells (2±2% vs. 36±10%). Similar findings were observed for engraftment of CD34+ cells. To enhance the migration of cytokine-cultured cells after IBMT, we similarly transplanted cultured AC133+ cells into the bone marrow of the left tibia, assessed the engraftment in the injected and noninjected tibiae at 7 days after transplantation, and then subcutaneously administered G-CSF (250 μg/kg/d) for 5 days. Administration of G-CSF stimulated the migration of cytokine-cultured cells to the bone marrow of previously-aspirated right tibia but failed to induce their migration to intact bone marrow of femur. These data indicate that ex vivo manipulation of hematopoietic progenitor/stem cells adversely influences their migration properties to other bone marrow compartments after IBMT. Our data raise caution for future clinical applications of the IBMT method using ex vivo-manipulated hematopoietic stem cells.
Allogeneic Committed Hematopoietic Progenitors Are Protective Against Radiation.
Authors: Chen Benny J. , Deoliveira Divino , Chao Nelson J. 
Published: October 2019

Abstract
Whole-body irradiation may lead to bone marrow failure and death. It was previously reported that congenic myeloerythroid-restricted progenitors are able to radioprotect lethally irradiated animals. However, this approach will not be practical because syngeneic/congenic donors are rarely available in humans. To solve this problem, we investigated whether allogeneic committed progenitors are also radioprotective. Hematopoietic committed progenitors were isolated by FACS based on the presence of early progenitor marker CD244 and the absence of stem cell marker CD150 (CD244+CD150−). BALB/c mice (H2d) were lethally irradiated with 8.5 Gy. Within 4 hours of irradiation, the irradiated mice were infused with 5x105 sorted hematopoietic progenitors from major histocompatibility complex mistmatched C57BL/6 donors (H2b). As shown in the Figure B, all the mice in the radiation control group died within 15 days post irradiation (median survival time: 13 days). Infusion of hematopoietic committed progenitors significantly prolonged the survival of the lethally irradiated mice (P=0.0018, median survival time: 28 days). These results are similar to the results obtained from congenic hematopoietic progenitors using 1x105 cells (Figure A, P<0.0001, median survival time: 10 days vs. 28 days). These data suggest that allogeneic hematopoietic committed progenitor cells are also able to mediate radioprotective effects. Similar to the congenic hematopoietic committed progenitors, allogeneic progenitors may also exert radioprotective effects by jumpstarting hematologic recovery post irradiation. These cells may be stockpiled and used as “off-the-shelf” products for radiation injury and other applications.
Figure Figure
Immunosuppressant Sinomenine Promotes Differentiation of Bone Marrow Progenitors to IL-10-Producing Ialow Regulatory Dendritic Cells Leading to the Prolongation of Allograft Survival.
Authors: Wang Quanxing , Wu Jianchun , Wang Jianli , Liu Yushan , Chen Guoyou , Cao Xuetao 
Published: October 2019

Abstract
Sinomenine (SN), an immunosuppressive compound derived from the Chinese medicinal plant Sinomenium acutum, effectively inhibits both proinflammatory factor production and lymphocyte proliferation. However, the effects of SN on dendritic cells (DC), the professional antigen-presenting cells, have not been elucidated. In this study, we examined the effects of SN on DC generation, maturation and function. We found that SN affected DC in a dose dependent manner; significantly inhibiting surface expression of MHC class II, CD86, and CD40, production of IL-12, TNF-a, IL-1b, and T cell-allostimulatory activity. SN targets antigen-presenting capacity of DC not only via the MHC class II pathway, but also impairs DC migration by inhibiting LPS-induced up-regulation of CCR7 and CXCR4 expression. These inhibitory effects on DC may be partially due to SN-mediated suppression of NF-kB and p38 MAPK pathways. Further study showed that SN-treated DC (DC-SN) induce donor-specific T cell hyporesponsiveness and trigger generation of IL-10-producing T regulatory-like cells in vitro. In vivo transplant studies revealed that, compared to pretreatment with immature DC, pretreatment of recipients with DC-SN could significantly prolong allograft survival, although long-term allograft acceptance was not achieved. Importantly, pretreatment of recipients with DC-SN in combination with SN administration profoundly prolonged allograft survival, resulting in long-term survival of 30% of allografts. Increased generation of CD4+CD25+ Treg cells and enhanced microchimerism in recipients brought on by combination treatment may contribute to this extension of allograft lifespan. Our results demonstrate that SN can promote bone marrow progenitors differentiation to IL-10-producing (Ialow) regulatory DC which contribute to induction of tolerance by increase generation of Treg cells.
Development of Tools for T Cell Repertoire Analysis (TCRB Spectratyping) for the Canine Model of Hematopoietic Cell Transplantation.
Authors: Kiianitsa Konstantin , Lesnikov Vladimir , Jordan Robert , Georges George E. 
Published: October 2019

Abstract
Analysis of the recombinatorial diversity of rearranged T cell receptor genes in mature T cells is an essential tool in the evaluation of immune status and immune reconstitution in hematopoietic cell transplantation studies. While the spectratyping technique is available in human clinical research and mouse models, the canine genome has not been sufficiently annotated to simply implement the assay on the dog TCRB locus. To this end, we have annotated by bioinformatics and experimentally all canine TCRBV segments, as well as TCRBD, TCRBC and most of TCRBJ segments. Of all 31 canine TCRBV found, 23 were functional and 8 were pseudogenes. A multiplex PCR-based assay was further designed to analyze the entire TCRBV spectratype in a set of 4 reactions each containing 4 to 5 V-segment specific forward primers and a common C-segment specific reverse primer. Direct sequencing of RT-PCR products confirmed that all amplified genes originated from predicted V-segments and that the designed V-specific PCR primers did not cross-react with other TCRBV family members. The usefulness of the spectratyping technique for canine model of transplantation was further demonstrated in analysis of T-cell repertoire reconstitution of irradiated dogs at different time points of recovery. Moreover, our simple and rapid V (J) annotation strategy relies on internet resources open to the general public and does not require specialized training in bioinformatics. It can be readily applied for de novo identification and mapping of TCRB gene families in other animal species where genome sequence drafts become available.
Figure 1. TCRB spectratype of fetal canine thymus. (A) RT-PCR using TCRBV family-specific forward primers (V1 through V26) and a common C region-specific reverse primer. (B) Amplification products specific for family V1 (high abundance) and V26 (low abundance) were copied in run-off reactions with the fluorescent C-specific primer and resolved on capillary gel. Figure 1. TCRB spectratype of fetal canine thymus. (A) RT-PCR using TCRBV family-specific forward primers (V1 through V26) and a common C region-specific reverse primer. (B) Amplification products specific for family V1 (high abundance) and V26 (low abundance) were copied in run-off reactions with the fluorescent C-specific primer and resolved on capillary gel.
A Vector Expressing PML-RARα Fused to GM-CSF Is an Effective DNA Vaccine for Inducing Specific Immune Response to APL Cells.
Authors: Li Yangqiu , Cen Dongzhi , Hu Gang , Zhou Yubing , Chen Shaohua , Yang Lijian , Przybylski Grzegorz K. , Schmidt Christian A. 
Published: October 2019

Abstract
Despite combinations of different therapeutic strategies have significantly increased survival, acute leukemia is still not curable. To further improve outcome, specific immunotherapy might be one of the best choice. DNA vaccines have been showed leading to strong and persistent cell-mediated and humoral immune response to the antigen encoded by the plasmid. However, little data exist regarding the DNA vaccines in acute leukemia, there are few studies reported that PML-RARα DNA vaccines were developed, but the host immune response were weakly, due to the weak immunogenicity of tumor antigens. In order to improve the effect of DNA vaccine for acute promyelocytic leukemia (APL) therapy, we have used a full-length human GM-CSF (hGM-CSF) sequence fused to PML-RARα breakpoint-drived sequence and develop a vector coexpressing PML-RARα gene and hGM-CSF gene, which was expected to to promote T cells response in host. PML-RARα fusion gene segment and the hGM-CSF gene were amplified from NB4 cells or pORF-hGM-CSF plasmid. Both PCR products were cloned into PIRES plasmid respectively to construct a recombinant plasmid PML-RARα-IRES-hGM-CSF. The recombinant plasmids were then transfected into K562 or A549 cells respectively. The expression of the PML-RARα/GM-CSF mRNA and protein in transfected cells were identified by RT-PCR, dot blotting, ELISA and Western-Blot respectively. By in vivo assays, BALB/c mice were vaccinated at 6–8 week of age with a total of 200 μg DNA in normal saline, injected into two sites in the quadriceps muscles on day 0, 7 and 21. The plasmid containing the same PML-RARα segment and blank plasmid served as controls. Two weeks after the final DNA boost, both PML-RARα/GM-CSF mRNA and protein, serum INF-γ and anti-NB4 cells specific cytotoxicity of splenocytes following 7 days of stimulation in vitro with freeze thawing NB4 cells and recombinant human IL-2 were assessed by ELISA and LDH assays. The results showed that the sequence of the fragments inserted in multi-clone site (MCS) A and MCS B of PIRES plasmid were absolutely correct by double restriction enzyme cutting analysis (Xba I/Sal I) and sequence analysis, the PML-RARα/GM-CSF mRNA and protein could be identified in transfected K562 or A549 cells and in mice quadriceps muscles. The level of serum INF-γ and cytotoxicity of splenocytes against NB4 cells from immunized mice was significant increased than that from control groups. In conclusions, the vector expressing PML-RARα and hGM-CSF was successfully constructed, which can more effective immune response and anti-APL cells effect in animal models than that from plasmid containing single PML-RARα segment. It could be farther used in the research as PML-RARα DNA vaccine for APL.
IFN-g-Stimulated Mesenchymal Stromal Cells Acquire the Ability To Cross-Present Exogenous Soluble Antigens and Lead to an Effective Immune Response.
Authors: François Moïra , Romieu-Mourez Raphaëlle , Stock-Martineau Sophie , Galipeau Jacques 
Published: October 2019

Abstract
Antigen cross-presentation is the mechanism by which exogenous antigens can be presented by major histocompatibility complex (MHC) class I molecules to CD8+ T cells. This process is efficiently performed by professional antigen presenting cells (APC) such as dendritic cells and macrophages. Recently, we and others have shown that IFN-g enables the upregulation of the expression of MHC class I & II molecules by marrow-derived Mesenchymal Stromal Cells (MSCs) and that MHC II-mediated antigen presentation can lead to cell-mediated protective immunity to xenoantigens [Stagg et al., Blood March 2006]. These findings led us to investigate whether MSCs also possess the ability to cross-present antigens via MHC class I. Using antigen presentation assays performed on murine MSC in the presence of MHC class I-restricted ovalbumin (OVA)-specific CD8+ T hybridoma cells or purified primary CD8+ T lymphocytes from OT1 transgenic mice, we demonstrated that MSC can robustly and effectively cross-present exogenous antigens via MHC class I molecules upon IFN-g pre-treatment in a manner comparable to professional APCs like macrophages. Use of transporter associated with antigen processing (TAP-1)-deficient mice and proteasome inhibitors suggested a MHC class I machinery-dependent pathway. Cross-presentation by IFN-g-activated MSC was also observed to be suppressed by TGF-b and regulated by cell density. In vivo, IFN-g-treated, OVA-pulsed MSCs administered to normal C57Bl/6 mice led to an effective OVA-specific, T cell cytotoxic immune response, leading to the rejection of OVA-expressing EG7 lymphoma cells. In conclusion, our findings suggest that cross-presentation properties of MSC could play a role in their effectiveness as conditional APCs in vivo and this property may be exploited as a therapeutic cell-based immune biopharmaceutical for treatment of cancer or infectious disease.