bluuebunny/crossref_metadata_2025_split huggingface dataset:
Autoantibodies Against p53 Are Associated with p17 Deletions in CLL.
Authors: Messmer Bradley T. , Nour-Omid Talia S. , Ghia Emanuela , Rassenti Laura , Kipps Thomas J. 
Published: April 2020

Abstract
The TP53 gene located on the short arm of chromosome 17 (17p) encodes p53, which is a transcription factor that regulates the cell cycle and the apoptotic response to genotoxic stress, such as that caused by most anti-cancer drugs. Loss of p53 function in chronic lymphocytic leukemia (CLL) typically is associated with deletions in 17p, which is associated with a poor response to many anti-cancer drugs and poor survival. Although only 5–10% of CLL patients at diagnosis have deletions in 17p by interphase fluorescence in situ hybridization (FISH), the frequency of cases that have 17p- increases in patients with disease that is refractory to chemotherapy. Assays that can screen for loss of p53 function in CLL could help identify patients who most likely will not respond well to most common forms of chemotherapy. Autoantibodies against p53 protein have been observed in several cancers and are, in some cases, linked to p53 protein accumulation in the tumor cells owing to loss of p53-induced expression of the p53-repressor MDM2. We examined the serum samples derived from platelet-free plasma of patients with CLL for anti-p53 autoantibodies by solid-phase ELISA. Anti-p53 autoantibodies were detected in 19 out of 31 plasma samples from patients with known 17p deletions defined by interphase FISH. In contrast, we detected anti-p53 autoantibodies in only 1 out of 27 plasma samples from patients with 11q22–23 deletions and in none of the 29 samples from patients with normal cytogenetics. Detectable levels of soluble p53 protein were identified in only 5 of the 31 patients with 17p deletions, and 3 of those had detectable anti-p53 antibodies. There was no correlation between anti-p53 autoantibody levels and the total plasma IgG. Samples from multiple time points were available for 22 of the patients with 17p deletions. In only one case did the p53 autoantibody status change (from positive to negative). The mechanism that leads to the generation of anti-p53 autoantibodies is unknown. One model suggests that constitutive or over expression of p53 protein by the leukemic cells may lead to a break in tolerance, perhaps if coupled with necrotic or otherwise immunogenic death of the cells. Studies are currently underway to address the status and function of the p53 protein in those patents with high autoantibody levels. This work suggests that auto-antibodies may be useful as a surrogate marker for p53 dysfunction and, more broadly, raises the possibility that auto-antibody profiles may provide clinically relevant insight into the biology and pathophysiology of CLL.
Gene Expression Signature of B-Cell Chronic Lymphocytic Leukemia with Trisomy 12.
Authors: Porpaczy Edit , Bilban Martin , Kroemer Elisabeth , Heinze Georg , Gruber Michaela , Vanura Katrina , Heintel Daniel , Hilgarth Martin , Hilgarth Bernadette , Le Trang , Hubmann Rainer , Schnabl Susanne , Chott Andreas , Schwarzinger Ilse , Hauswirth Alexander , Scrabs Cathrin , Stilgenbauer Stephan , Wagner Oswald , Fonatsch Christa , Jaeger Ulrich , Shehata Medhat , Gaiger Alexander 
Published: April 2020

Abstract
The prognosis of patients with B-CLL is largely determined by the karyotype of the malignant clone. Microarray technology has facilitated linkage between chromosomal aberrations and gene expression signatures. We have investigated the gene expression profile associated with trisomy 12 (+12). Expression data were obtained by microarray analysis of mRNA from unselected PBMNC of 4 patients with +12 and compared with 16 B-CLL controls. 146 genes were at least 2-fold over- or underexpressed in samples with +12. Five of the 16 genes showing the strongest correlation with +12 were selected for further analysis (HIP1R FC=3,43; MYF6 FC=3,92; P2RY14 FC=−9,59; RASGRP3 FC=−3,85; SLC2A6 FC=2,13) and validation by real time PCR: HIP1R located on chromosome 12q24, with a fold change (FC) of 3,43, MYF6 (chromosome 12q21, FC=3,92), P2RY14 (chromosome 3q21–q25, FC-9,59) RASGRP3 (chromosome 2p25.1–p24.1, FC=−3,85). SLC2A6 (chromosome 9q34, FC=2,13). Quantitative PCR was performed with mRNA from 61 patients (29 with +12, 32 B-CLL controls) and 2 healthy donors. Only 3 genes were significantly associated with +12 compared to the B-CLL-controls in this evaluation: HIP1R (3,486; p&lt;0,0001), MYF6 (1,498; p=0,005), P2RY14 (1,216; p=0,013). (Table1). Two of these genes (HIP1R, MYF6) are located on chromosome 12 indicating a “gene dosage effect”, while P2RY14 is localized on a different chromosome suggesting trans-acting processes. We have used expression of HIP1R as a surrogate marker for trisomy 12. The predicted sensitivity was 79,3% and the predicted specifity was 90,6. Analysis of CD19+ selected B-CLL and normal B-cells revealed that MYF6 is exclusively expressed in normal or malignant B-cells in peripheral blood. We confirmed that MYF6 is highly specific for skeletal muscle, however strong expression was found in normal tonsils, DLBCL, and other B-cell malignancies. Our data link a specific gene expression signature with trisomy 12. 3 novel marker genes were identified, which could be used as diagnostic tools. The linkage with P2RY14 suggests that +12 influences the expression of genes from other chromosomes.
Table 1 Mean + 12, N=29 Mean B-CLL controls, N=32 p-value Locus Fold change microarray HIP1R 0,7819 0,2243 0,000 12q24 3,43 MYF6 37,55 25,06 0,005 12q21 3,92 P2RY14 −0,2873 0,3494 0,013 3q21–q25 −9,59 RASGRP3 0,8404 1,0774 0,055 2p25.1–p24.1 −3,85 SLC2A6 29,78 22,37 0,080 9q34 2,13
The Development of Severe Anti-HPA 1a-Related Neonatal Alloimmune Thrombocytopenia Is Influenced by the Maternal ABO Type.
Authors: Ahlen Maria T. , Killie Mette K. , Husebekk Anne , Kjeldsen-Kragh Jens , Olsson Martin L. , Skogen Bjorn 
Published: April 2020

Abstract
About 100 000 pregnant women were included in a screening and intervention program aimed to reduce morbidity and mortality related to neonatal alloimmune thrombocytopenia (NAIT). NAIT due to HPA 1a immunization have many features in common with hemolytic disease of the newborn (HDN) caused by RhD immunization. Platelets express blood group A and B antigens, and as described in HDN, ABO incompatibility between mother and fetus could possibly protect against immunization against HPA 1a. The aim of the study was to describe the ABO distribution among the immunized mothers and examine whether ABO status of the mother influenced the severity of thrombocytopenia in the newborn. ABO typing of HPA 1a immunized women was routinely performed by serological methods during the screening. ABO genotyping resolving the major alleles (A1, A2, B, O1, O1v or O2 alleles) of the women and their newborns was performed by PCR-RFLP methods. The ABO distribution among the immunized HPA 1a negative women was found to be similar to the normal distribution in the Norwegian population, which may indicate that the ABO type does not influence the immunization mechanism. However, we find that HPA 1a immunized women of blood group A have a higher risk of delivering a child with severe NAIT (platelet count &lt; 50×109/L) than women with blood group O. Twenty percent of the immunized women with blood group O gave birth to children with severe NAIT, compared to 46% among the immunized blood group A mothers, resulting in a relative risk of 0.43 (95% CI 0.25–0.76). The ABO type of the newborn was not found to influence development of severe NAIT. The O1/O1v allele distribution among the immunized women with blood group O resembles the distribution reported for a Swedish population. However, only 2/22 (9.1%) pregnancies among the O1v negative blood group O mothers resulted in a newborn with severe NAIT, compared to 10/34 (29.4%) among the O1v-positive blood group O women, resulting in a relative risk of 0.31 (95% CI 0.07–1.28). The observation that the immune response against HPA 1a may have different consequences depending on the ABO blood group of the mother is interesting. O1v, also termed O02, constitutes a separate but ancient allelic lineage at the ABO locus and we are now in the process of examining if our observation is related to linkage disequilibrium between the ABO gene and one or more pregnancy-related immunoregulatory genes.
Bleeding Complications and Pregnancies in an Established Plasminogen Activator Inhibitor, Type-1 (PAI-1) Deficient Population.
Authors: Mehta Rakesh P. , Simmons Molly , Heiman Meadow , Shapiro Amy D. 
Published: April 2020

Abstract
In 1992, a young Amish girl with marked post-procedural bleeding was found to have a PAI-1 deficiency with a homozygous frame shift mutation in exon 4 of the PAI-1 gene that results in a non-functional protein. This mutation was tracked within the ancestors of this Amish community and two individuals were identified that likely introduced this gene into the community. In 1997, seven children (including the proband) from two families within the kindred were described with symptoms consistent with a mild bleeding disorder. Two further homozygous children, one from a third family within the kindred, have subsequently been identified. Presently, over 300 members of this kindred have been tested with nearly 100 identified heterozygotes, none of whom have experienced bleeding complications. The homozygotes all have had bleeding problems with injury a common cause. Five of the nine homozygotes have had head trauma, resulting in significant bleeding complications. ε-aminocaproic acid (EACA) initiated early controls bleeding. One child had a recurrence of his subdural hematoma when he discontinued the EACA upon discharge from the hospital. But the bleeding was subsequently controlled with reinstitution of EACA, and the child had a full recovery. Another common bleeding complication is menorrhagia with all homozygous females experiencing this symptom, and two even requiring red blood transfusions due to severe anemia. Currently, these women require concurrent EACA with onset of menses to control bleeding. Of interest PAI-1 and the plasminogen activation system have been shown to play an important role in embryo implantation, and it was unknown whether homozygous PAI-1 deficient women would be able to conceive and maintain a pregnancy. We now report 2 of these homozygous deficient females have become pregnant; one remains pregnant at 28 weeks gestation, and one delivered at 32 weeks. This child represents that first documented birth to a woman with complete PAI-1 deficiency. The mother experienced abnormal cervical bleeding beginning at 27 weeks requiring EACA therapy which was continued until delivery and for about 7 days post-partum without abnormal bleeding at delivery or in the post-partum period, and without ill effect on the fetus. This cohort is the largest group of PAI-1 deficient patients ever documented. The molecular defect is well defined so that it is simple to differentiate normals from heterozygotes and homozygotes. Continued follow-up will allow further understanding of the role of PAI-1 in human physiology.
Histone Deacetylase Inhibitor SAHA Reduces Cytokine Storm and Facilitates Induction of Chimerism When Combined with Anti-CD3 mAb in Conditioning of Recipients.
Authors: Li Nainong , Zhao Dongchang , Zhang Chunyan , Lin Chia-Lei , Kirschbaum Mark , Kandeel Fouad , Forman Stephen , Zeng Defu 
Published: April 2020

Abstract
Donor T cell-mediated graft versus leukemia (GVL) and graft versus autoimmunity (GVA) plays a critical role in the treatment of hematological malignancies and refractory autoimmune diseases using allogeneic hematopoietic cell transplantation (HCT). However, graft versus host disease (GVHD) remains a major obstacle in classical HCT, where recipients are usually conditioned with total body irradiation or high dose chemotherapy. We have recently reported a radiation-free and GVHD preventive anti-CD3-conditioning regimen, which allows donor CD8+ T cells to facilitate engraftment and mediate GVL without causing GVHD (PNAS, 2007 and J. I. 2007). In order to promote the clinical application of the anti-CD3-conditioning regimen, we need to overcome the cytokine storm that causes flu-like syndrome in patients after anti-CD3 mAb injection and reduce the required donor bone marrow cell dose. Suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, was reported by others to induce apoptosis of human T cell lymphoma cell lines and to reduce serum levels of pro-inflammatory cytokines in HCT recipients. Therefore, we tested whether conditioning with a combination of anti-CD3 and SAHA can reduce the cytokine storm caused by anti-CD3 and reduce the resistance of engraftment mediated by residual host T cells. Accordingly, titrated dose of SAHA (0.125–8 μM) were added to cultures of BALB/c spleen cells stimulated with plate-bound anti-CD3. We found that SAHA augmented apoptosis of anti-CD3 activated T cells in a dose-dependent manner. Although low-dose SAHA (0.125–0.25 μM) did not augment apoptosis, it rendered the residual live T cells partially unresponsive. At the same time, SAHA significantly reduced the pro-inflammatory cytokines (IL-2, IFN-γ, TNF-α, and IL-6) in the culture supernatants. Similarly, in vivo treatment with anti-CD3 and low-dose SAHA (40 μg/g) led to significant reduction of serum levels of those cytokines and partial unresponsiveness of the residual host T cells. Finally, conditioning with combined anti-CD3 and SAHA (40 μg/g) induced complete chimerism without GVHD in 12/12 of old BALB/c as well as in 8/8 of old autoimmune lupus NZB/NZW F1 recipients after two injections of BM cells (2 ×106/g) and CD4+ T-depleted spleen cells (4 ×106/g), although conditioning with anti-CD3 alone did not induce any chimerism (0/8). The chimeric NZB/NZW F1 recipients showed complete reversal of autoimmune glomerulonephritis and proteinuria. These results indicate that SAHA can not only reduce cytokine storm but also facilitate engraftment when combined with anti-CD3 for conditioning of recipients. This radiation free and GVHD preventive conditioning regimen may provide a novel approach for clinical HCT.
CD6-Depletion in the Dog Model: New Insights into Chimerism and Tolerance.
Authors: Zorn Julia , Herber Martin , Schwamberger Sabine , Adler Heiko , Kolb Hans-Jochem 
Published: April 2020

Abstract
Adoptive immunotherapy with donor lymphocyte infusion (DLI) following bone marrow transplantation may produce a graft-versus-leukemia (GVL) effect after transplantation of T-cell-depleted bone marrow. However, this method is limited by a high risk of GvHD. We studied the occurrence of graft-versus-host disease (GVHD) and chimerism after DLI at various times after CD6-depleted bone marrow transplantation in a canine model of DLA-homozygous donor and DLA-heterozygous littermate recipient. Using a cross reactive mouse-anti-human antibody which recognizes the canine CD6 antigen, a simple and economical depletion method could be established, which makes use of the ability of the CD6 antibody MT-606 to activate the complement cascade. In vitro experiments using MT-606 and rabbit complement showed the effective depletion of CD6-positive cells whereas hematopoietic progenitor cells remained unaffected at the same time. For in vivo studies, DLA heterozygous recipients with one DLA haplotype being identical to the donor were treated with 10 Gy total body irradiation (TBI) and received CD6-depleted bone marrow of their DLA-homozygous littermate donors. In the control group 7 dogs given unmanipulated marrow died of GVHD within 28 days of transplantation. Two dogs showed sustained engraftment without the occurrence of GvHD after the infusion of 1,8 · 108/kg – 2,0· 108/kg CD6-depleted mononuclear marrow cells. The establishment of tolerance to the bone marrow donor was shown by the transplantation of skin from the donor to the host. Transfusion of donor lymphocytes (1.0 · 108 MNCs/kg body weight) on day 3, 7 or 14 induced GvHD in the host. However, with DLI on day 20, only 2 out of 4 dogs developed GvHD, but DLI always converted mixed into complete chimerism. We conclude from this study that the cross reactive CD6 antibody MT606 effectively depletes T cells in the presence of rabbit complement and induces GVH tolerance without jeopardizing engraftment, and the risk of GVHD after DLI is reduced with time after transplantation.
Host Interferon gamma Production and STAT-1 Signalling Is Crucial for Minor Antigen Mediated Rejection of High Grade Lymphoma.
Authors: Sukumar Madhusudhanan , Wilke Andrea , Jaeger Eva , Mautner Josef , Ellwart Joachim , Kolb Hans-Jochem , Bornkamm Georg W. , Gerbitz Armin 
Published: April 2020

Abstract
To date mechanisms of T-cell mediated immunity and immune-escape in high grade lymphomas are poorly understood. Using a transgenic mouse lymphoma model, where the proto-oncogene c-myc is driven in parts of the immunoglobulin lambda locus representing a t(8;22) translocation as found in Burkitt’s lymphoma, we developed a syngeneic model to investigate the anti-lymphoma activity of lymphoma specific T-cells. By retroviral transduction of a lymphoma specific antigen (chicken ovalbumin-IRES-GFP vector) into primary cell lines from c-myc transgenic lymphomas we established a model that would allow us to investigate the contribution of interferon gamma signalling in rejection of high grade lymphoma. All lymphomas established displayed low MHC class I and II levels on the surface when compared to wildtype B-cells. This expression could be enhanced by treatment with interferon gamma (100U/ml) up to 10 fold. When retrovirally transduced lymphoma cells were injected into wildtype or GFP transgenic C57BL/6 recipients, animals displayed a significant delay in lymphoma growth compared to IRES-GFP transduced control cell lines. 50% of the recipient mice rejected OVA containing lymphomas whereas we observed a 100% lymphoma growth of IRES-GFP transduced lymphomas in GFP transgenic recipients. Developing OVA containing lymphomas displayed a loss of GFP expression indicating a selection for non transduced cells. In spleens from mice successfully rejecting OVA-containing lymphomas we found up to 1.5% (±0.12%) SIINFEKL specific T-cells. To gain mechanistic insights of lymphoma rejection, we transferred OVA transduced lymphoma cells to Stat1−/− and IFNg−/− recipients. Lack of STAT1−/− on the recipient side or inability to secrete interferon gamma was associated with fast lymphoma progression and was not different when compared to IRES-GFP transduced cell lines injected into GFP transgenic hosts. Although we found 1.6% (±0.52%) SIINFEKL T cells in spleens of lymphoma bearing STAT1−/− animals, interferon gamma production was significantly decreased. We could also show that in wild type recipients, OVA containing lymphomas displayed high MHC class I and II expression which is completely absent in lymphomas from Stat1−/− recipients. Our results suggest that rejection of high grade lymphoma by a specific minor antigen is possible and Stat1 signaling and interferon gamma production by the host is crucial for lymphoma rejection.
CD62L Is Not Critical for T Regulatory Cell-Mediated Protection from Lethal Acute GVHD.
Authors: Carlson Michael J. , Siedel Hans E. , Serody Jonathan S. 
Published: April 2020

Abstract
INTRODUCTION: Graft-versus-host disease (GVHD) is a major complication following allogeneic bone marrow transplantation. Despite advances in understanding the etiology of GVHD it remains a formidable obstacle to the widespread application of BMT. T regulatory (Treg) cells have been shown to inhibit GVHD while preserving the beneficial graft-versus-leukemia (GVL) effect. Published studies by numerous groups, including our own, have shown the importance of Treg homing molecule expression in preventing GVHD. Of note, the Treg population that expressed high levels of the adhesion molecule L-Selectin (CD62L) was shown to be capable of promoting BM engraftment and inhibiting lethal acute GVHD, whereas the CD62Llo population was not. While these studies provided evidence that the phenotype of the CD62Lhi Treg population was responsible for inhibition of GVHD they did not directly assess the role of CD62L.
METHODS: To investigate the importance of CD62L in Treg mediated protection from GVHD we utilized a well-described parent into F1 model. We transferred Tregs (2×106) from CD62L deficient mice (CD62L−/−) or wild type (WT) C57BL/6 mice with WT naïve T cell effectors (5×106) into lethally irradiated B6D2 recipients. Our results demonstrated similar survival of recipient mice receiving CD62L−/− Tregs (40% survival) compared to those receiving WT Tregs (60% survival: p =0.66). In addition fluorescence microscopy revealed no difference in GFP+ T cell effector migration/accumulation in GVHD target organs (liver, lung, GI tract) or secondary lymphoid organs (spleen, mesenteric lymph node, inguinal lymph node, Peyers Patches) in the presence of CD62L−/− or WT Tregs. Also, we tracked the migration of GFP+ CD62L−/− Tregs in vivo by fluorescence microscopy and found they were able to efficiently migrate to secondary lymphoid organs and GVHD target organs. Surprisingly, GFP+ CD62L−/− Tregs were also able to home to peripheral lymph nodes.
CONCLUSIONS: These results suggest CD62L is dispensable for Treg mediated suppression of GVHD and that the function of the L-selectinhi Treg fraction may be due to the difference in the expression of other homing/migration molecules necessary for Treg migration into lymphoid and GVHD target organs.
Figure Figure
Cbl Mutants Interact with c-Kit and Lead to Factor Independent Growth and Transformation of a Myeloid Cell Line 32D.
Authors: Sargin Bülent , Bandi Srinivasa Rao , August Benjamin , Rensinghoff Marion , Choudhary Chunaram , Gonchi Pavan Nagaruri , Berdel Wolfgang E. , Müller-Tidow Carsten , Serve Hubert 
Published: April 2020

Abstract
The Cbl proto-oncogene products have emerged as important components of the signal transduction cascades downstream of both non-receptor and receptor tyrosine kinases (RTKs). By regulation of receptor trafficking and degradation, they have been shown to tightly regulate the intensity and amplitude of RTK activation. c-Kit belongs to the family of the class-III RTKs and plays an important role in the pathogenesis of acute myeloid leukemia (AML). So far, very little is known about the role of c-Cbl mutants in the role of c-Kit signaling. We analyzed the interaction of c-Cbl with c-Kit and the functional relevance of this interaction in the IL-3-dependent murine myeloid progenitor cell line 32Dcl3. We analyzed the effect of c-Cbl and two different dominant negative mutants of c-Cbl (Cbl-70Z and Cbl-R420Q) on c-Kit-ligand-activated internalization. The transfection of c-Cbl mutants, but not of wildtype c-Cbl, significantly inhibited receptor internalization, as analyzed by FACS analysis. Expression of Cbl-70Z in 32Dcl3 cells severely inhibited apoptosis induced by growth factor deprivation, as has been described before. However, when coexpressed with wildtype c-Kit, 32Dcl3 cells also rapidly proliferated in the absence of any exogenously added growth factors. We furthermore analyzed SCF induced c-Kit ubiquitination in the presence and absence of c-Cbl mutants. SCF induced rapid ubiquitination of c-Kit that was strongly reduced in the presence of Cbl-70Z and abolished by Cbl-R420Q. Also, the Cbl mutants altered the amplitude and changed the quality of c-Kit dependent signaling events. In colony assays we were able to show ligand independent colony growths in methyl cellulose only in the presence of wildtype c-Kit together with a c-Cbl mutant. Our results indicate that c-Cbl has an important role in c-Kit signal mitigation. Furthermore, they demonstrate that disturbed mechanisms of c-Kit internalization have important implications for its transforming potential, possibly in the development of AML.