bluuebunny/crossref_metadata_2025_split huggingface dataset:
Murine Ultrasonography Confirms That Rapamycin Is Effective Against Autoimmune Lymphoproliferative Syndrome (ALPS).
Authors: Teachey David T. , Axsom Kelly , Brown Valerie I. , Obzut Dana , Grupp Stephan A. 
Published: April 2020

Abstract
Autoimmune Lymphoproliferative Syndrome (ALPS) is a disorder of abnormal lymphocyte survival caused by defective Fas-mediated apoptosis. Patients with ALPS develop lymphadenopathy, hepatosplenomegaly, and an increased number of an unusual T cell population, characterized as T cells that are CD3+ CD4− CD8− and referred to as double negative T cells (DNTs). Although ALPS is a rare disorder, we have shown it may be more common than previously appreciated. Treatment options for patients with ALPS are limited. Rapamycin, an mTOR inhibitor, has been shown to induce apoptosis in nonmalignant B and T lymphocytes and we have demonstrated its potential efficacy in models of lymphoid leukemia. Because ALPS is caused by defective lymphocyte apoptosis, we hypothesized rapamycin would be effective in ALPS by inducing apoptosis in these abnormal cells, controlling the lymphoproliferation that is the hallmark of the disease. We confirmed this hypothesis by using a murine model of ALPS, CBA-lprcg, treating animals that had developed significant lymphadenopathy and splenomegaly with rapamycin. We followed response to treatment with serial assessment of DNTs in peripheral blood and lymphoid tissue by flow cytometric determination and by serial monitoring of lymph node and spleen size with small animal ultrasound. This is the first report to use this novel and powerful tool to measure murine lymphoproliferation and we found 3-dimensional ultrasound measurements in the mice correlated to actual tissue measurements at sacrifice (r = 0.9648, p = 0.0001). We found a dramatic and statistically significant decrease in DNTs (p = 0.02), lymphadenopathy (p = 0.003), and splenomegaly (p = 0.03) after only 4 weeks when comparing mice treated with rapamycin to control animals (Table). We found no toxicities and no evidence of myelosuppression in treated animals. We performed immunoblots to evaluate a major mTOR signaling pathway intermediate (phospho-S6) on lymph node biopsies obtained from mice before and after treatment with rapamycin and found a correlation between clinical response and biochemical response (downregulation of phospho-S6). In summary, rapamycin, a safe and well tolerated medicine, is effective in murine ALPS and its use should be explored in patients with the disease.
Rapamycin Effective in ALPS Disease Parameter (1) Rapamycin Treated(2) Vehicle Control(2) p value (1) Average number of DNTs, volume of lymph nodes, and area of spleens were the same between groups at initiation of treatment. (2) Average (range) after 4 weeks of treatment DNTs of blood by FACS (mm3) 2440 (1343 – 3756) 9957 (4426 – 20384) 0.02 Lymph node volume by ultrasound (mm3) 52 (3 – 193) 760 (366 – 1043) 0.003 Splenic area by ultrasound (mm2) 54 (20 – 103) 179 (95 – 336) 0.03
Bortezomib Alters Peripheral Blood Lymphocyte Subsets in Patients with Multiple Myeloma.
Authors: Uy Geoffrey L. , Holt Matthew S. , Fisher Nicholas M. , Devine Steven M. , Tomasson Michael H. , DiPersio John F. , Vij Ravi 
Published: April 2020

Abstract
Bortezomib (VELCADE®) is a potent inhibitor of the proteasome which exerts its antimyeloma effect in part by blocking the activation of NF-κB. As NF-κB is critical for lymphocyte development and survival, there is great interest in harnessing the potential immunomodulatory effects of bortezomib. In murine hematopoietic transplantation models, bortezomib inhibits in vitro mixed lymphocyte responses and promotes the apoptosis of alloreactive T cells protecting against acute graft-versus-host disease. However, no data exists on the in vivo effects of bortezomib on human T cells. To characterize the effects of bortezomib on immune function, we profiled peripheral blood lymphocytes subsets and T cell associated cytokines in 39 patients with multiple myeloma. Two cycles of bortezomib 1.3 mg/m2 were administered by intravenous infusion on days 1, 4, 8, and 11 of a 21-day treatment cycle. The patients had received prior induction chemotherapy and would proceed to autologous transplant following treatment with bortezomib. Study population consists of 23 male and 16 female patients with the median age of 56 years (range 38–69). Myeloma characteristics at diagnosis were as follows (number of patients): IgG (28), IgA (10), light chain only (1), with stage I (1), II (12), or stage III (26) disease. Peripheral blood was collected at baseline (cycle 1, day 1) and at one week after the last dose of bortezomib (cycle 2, day 18) and analyzed for lymphocyte subsets by standard multicolor flow cytometry. Th1 and Th2 serum cytokines were measured at the same timepoints using a multiplexed cytometric bead array (BD Biosciences). Following treatment with bortezomib, no significant changes were detected in either Th1 or Th2 serum cytokine levels: IL-2 (p=0.116), TNF-alpha (p=0.854), IFN-gamma (p=0.070), IL-4 (p=0.240), IL-6 (0.236), IL-10 (0.151) as analyzed by Wilcoxon signed ranks test. Analysis of lymphocyte subsets using a paired student’s T-test demonstrated a 38% decrease in CD56+ NK cells (p=0.02) and a 26% increase in CD4/CD8 ratio (p=0.0006) which appears to be secondary to a decrease in CD8+ cytotoxic T-cells (p=0.054). (Table 1.) In conclusion, we observe an alteration of lymphocyte subsets following only two cycles of bortezomib. Further analysis of the effects of long term treatment with bortezomib is warranted. These studies may provide insights into the role of bortezomib as an immunomodulatory agent.
Peripheral Blood Lymphocyte Subsets Pre-bortezomib (/mm 3 ) Post-bortezomib(/mm 3 ) Difference(/mm 3 ) P-value CD2 1446 1259 −187 0.085 CD3 1273 1160 −113 0.28 CD4 842 802 −40 0.54 CD8 412 337 −75 0.055 CD19 90 94 4 0.86 CD20 87 95 8 0.78 CD56 206 148 −58 0.022 CD4/CD8 ratio 2.53 3.19 0.66 0.0006
Trafficking and Expansion of NK Cells in Murine Models of Bone Marrow Transplantation.
Authors: Olson Janelle A. , Negrin Robert 
Published: April 2020

Abstract
Allogeneic hematopoietic cell transplantation has proven to be an effective treatment for hematologic malignancies and some solid tumors. However, the high incidence of graft versus host disease (GVHD) as a complication of this treatment has limited the overall effectiveness of HCT. In addition, disease relapse also limits overall success. Novel strategies are needed which can suppress the development of GVHD but still maintain an effective immune response to provide a GVT effect. It has been shown that NK cells have the capability of suppressing the development of GVHD while inducing an anti-tumor response. The trafficking of cells to specific organs and tissue sites after transplant can be a major factor in determining whether or not they contribute to GVT and GVHD reactions. Little is known about the trafficking patterns of NK cells following hematopoietic cell transplantation, their proliferation or how long they persist in vivo. We investigated the trafficking patterns of NK cells in vivo in an allogeneic and syngeneic bone marrow transplant setting using a novel in vivo bioluminescence imaging (BLI) technique. Freshly isolated NK cells from FVB L2G85 transgenic mice, which constitutively express the luciferase gene and can be imaged by BLI, were transplanted along with T-cell depleted bone marrow into lethally irradiated BALB/c (allogeneic) or FVB (syngeneic) mice. BLI of the irradiated mice on successive days post-transplant indicated that in the allogeneic setting, NK cells traffic to distinct lymphoid organs, such as the cervical lymph nodes and spleen. The bioluminescent signal was 2.75-fold greater on Day 6 post-transplant than on Day 2 post-transplant, indicating a significant in vivo expansion of the NK cells in an allogeneic recipient in the first week post-transplant. In the syngeneic setting, NK cell trafficking to distinct lymphoid organs was not observed, and the bioluminescent signal intensity emitted from the transplanted mice remained constant. Thus, we anticipate that the in vivo expansion of NK cells seen in the allogeneic bone marrow transplant setting is not due to homeostatic proliferation alone. Continuing studies will address whether the in vivo expansion is driven by NK cell receptor-ligand interactions or MHC Class I differences. Understanding NK cell trafficking and proliferation could provide novel insights into enhancing function of both innate and adoptively transferred NK cells.
Vaccination of Advanced Prostate Cancer Patients with PSCA and PSA Peptide-Loaded Dendritic Cells Induces Cellular Immune Responses That Correlate with Superior Overall Survival.
Authors: Kaskel Anna K. , Zeiser Robert , Jochim Rosa , Schultze-Seemann Wolfgang , Waller Cornelius , Veelken Hendrik 
Published: April 2020

Abstract
Prostate stem cell antigen (PSCA) and prostate-specific antigen (PSA) are overexpressed in most prostate cancers. PSCA- and PSA-derived, HLA-A2 binding peptides are specific targets for T cell responses in vitro. A phase I/II trial was performed to demonstrate feasibility, safety, and induction of antigen-specific immunity by vaccination with dendritic cells (DC) presenting PSCA and PSA peptides in patients with hormone- and chemotherapy-refractory prostate cancer. Patients received four vaccinations with median of 2.7x10e7 peptide-loaded mature DC s.c. in biweekly intervals. Twelve patients completed vaccination without relevant toxicities. Six patients had stable disease (SD) after four vaccinations. One patient had a complete disappearance of lymphadenopathy by ultrasound despite rising PSA. Regarding PSA kinetics, in one patient, the log PSA slope became negative after treatment, and three patients had significant decreases in the log PSA slopes. However, duration of response was short, with a median time to progression of 65 days (12–223). Four patients with SD and one progressor developed a positive DTH after the 4th vaccination. With a median survival of all patients of 13.4 months, DTH-positivity was associated with significantly superior survival (22 vs. 8 months, p=0.003). HLA tetramer analysis detected high frequencies of peptide-specific T cells after two vaccinations in one patient who was also the sole responder to concomitant hepatitis B vaccination as an indicator of immune competence and survived 27 months after start of vaccination. Taken together, vaccination with PSA/PSCA peptide-loaded, autologous DCs may induce cellular responses primarily in immunocompetent patients which were associated with superior outcome. The retardation of PSA kinetics upon vaccination suggest a causal influence of the treatment on the natural course of the disease. Testing of DC-based vaccination is warranted for patients at earlier stages of prostate cancer.
Effect of Treatment with Rituximab in Patients with Refractory Autoimmune Cytopenias.
Authors: Dungarwalla Moez , Marsh Judith , Tooze Jenny , Lucas Geoff , Ouwehand Willem , Pettengell Ruth , Dearden Clare E. , Gordon-Smith Edward C. , Elebute Modupe O. 
Published: April 2020

Abstract
The autoimmune cytopenias encompass the disorders of immune thrombocytopenia purpura (ITP), autoimmune haemolytic anaemia (AIHA), autoimmune neutropenia (AIN), pure red cell aplasia (PRCA) and various combinations of these conditions. We describe 34 patients with severe autoimmune cytopenias resistant to standard immunosuppression treated with the anti-CD20 monoclonal antibody Rituximab. 17 patients had ITP, 6 AIN, 2 AIHA, 2 PRCA, 4 AIHA and ITP (Evan’s syndrome), 1 AIHA and AIN, 1 PRCA and ITP, and 1 AIN and ITP. 28 patients had primary autoimmune cytopenias and in 6 patients, the cytopenia was secondary to an underlying disorder (3 B-NHL, 1 T-NHL, 1 B-CLL and 1 SLE).
Rituximab was administered intravenously at a dose of 375mg/m2 weekly for 4 weeks. 33 of 34 patients completed treatment. In one patient treatment was discontinued after 3 infusions due to a hypotensive syncopal episode. No other major infusion related side effects were observed. Responses were seen in 9/17 patients with ITP, 2/2 patients with AIHA and 4/4 patients with AIHA and ITP. Responses were sustained in 8/9 ITP patients with median follow up of 25.8months (range 4–66 months). We observed a complete response in 5 of 7 ITP patients who were previously splenectomised.
In contrast, there was no response in 5/6 patients with AIN and in 3/3 patients with PRCA.
In two patients we observed severe neutropenia at 3 months post Rituximab which was associated with fatal infection in both patients, namely pnemocystis pneumoniae infection and acute exacerbation of bronchiectasis. Apart from these 2 cases no other infections were seen.
In conclusion, Rituximab appears to be a an alternative and safe option in the treatment of patients with ITP, AIHA and Evan’s syndrome inducing sustained clinical remissions in patients who have failed to respond to conventional immunosuppression and/or splenectomy. In contrast Rituximab does not seem to be effective in AIN and PRCA and this may reflect a different pathogenesis for these conditions.
Gene Expression Analysis Uncovers Possible Resistance Mechanisms to Bortezomib in Mantle Cell Lymphoma That Differ from Mechanisms Identified in Other Lymphoid Malignancies.
Authors: Rizzatti Edgar G. , Mora-Jensen Helena I. , Billings Eric , Lai Raymond , Wiestner Adrian 
Published: April 2020

Abstract
Bortezomib, a potent inhibitor of the 26S proteasome, has remarkable activity against some lymphoid malignancies, particularly multiple myeloma and mantle cell lymphoma (MCL). The observed 30–50% clinical response rate to bortezomib as a single-agent in relapsed MCL is believed to be mediated mainly by inhibition of the NFkB signaling pathway. Recently, Hsp27 has been shown to confer resistance to bortezomib in a lymphoid cell line. However, much less is known about the mechanisms underlying anti-tumor activity of or resistance to bortezomib in MCL. To address these questions we studied 10 MCL cell lines with t(11;14)(q13;q32) as in vitro models. IC50 values were measured by the MTT cytotoxicity assay for both bortezomib and the BAY 11-7082 compound, a specific inhibitor of the NFkB pathway. The cell lines showed different profiles of response to bortezomib and were grouped according to their IC50 values as sensitive (S) (Granta 519, Jeko-1, SP-49, UPN-1), intermediate (In) (HBL-2, JVM-2, Z-138) and resistant (R) (Mino, NCEB-1, SP-53). The mean IC50 for bortezomib in the R group was 3 times higher than the mean IC50 of the S group. The cell lines also showed very different profiles of response to the NFkB-specific inhibitor BAY 11-7082, but grouped differently, suggesting that other mechanisms, in addition to the NFkB pathway, participate in the anti-neoplastic effect of proteasome inhibition in MCL. Resistance to both drugs had no correlation with P-glycoprotein activity as measured by the rodhamine efflux assay. We performed gene expression profiling on Affymetrix U133A 2.0 arrays of all 10 cell lines. Using GeneSpring software (Agilent), we normalized expression to the mean of the S group and identified 79 transcripts that showed a dose response behavior; that is, genes whose mean expression was 1.5x higher in the In versus the S group and 1.5x higher in the R versus the In group. Conversely, 55 transcripts followed the opposite trend, being down-regulated at least 1.5x in both comparisons. This list contained genes important in apoptosis control and in transmembrane transport, but was most notable for an overexpression of stress response genes in the R group. Notably, expression of Hsp70 was almost 10-fold higher in the R versus S group. We therefore analyzed expression of the HSP70 as well as HSP27 and HSP90 proteins by Western blotting. Hsp70 protein was highly expressed in all MCL cell lines and there was no discernable correlation with bortezomib resistance for any of the HSP-proteins analyzed. Other resistance mechanisms appear more important in MCL and we are currently testing further candidate genes for their ability to confer resistance to bortezomib in MCL.
Co-Treatment with the hsp90 Inhibitor 17-Dimethylaminoethylamino-17-Demethoxygeldanamycin (DMAG) and Histone Deacetylase Inhibitor (HDI) Vorinostat (Suberoylanilide Hydroxamic Acid, SAHA): A Highly Active Combination Against Human Mantle Cell Lymphoma (MCL) Cells.
Authors: Hatter Angela , Bali Purva , Balasis Maria , Fiskus Warren , Boyapalle Sandhya , Kumaraswamy Sandhya , Rocha Kathy , Pranpat Michael , Hannah Alison , Sotomayor Eduardo , Richon Victoria , Bhalla Kapil 
Published: April 2020

Abstract
We have previously reported that agents that inhibit ATP binding and chaperone function of hsp90 are highly active against wild type and mutant Bcr-Abl and mutant FLT-3 containing human acute leukemia cells. In the present studies, we determined the effects of a more soluble and potent geldanamycin analogue, DMAG (Kosan Biosciences Inc.), and/or hydroxamate histone deacetylase inhibitor (HHDI), vorinostat (Merck & Co., Inc.), against human MCL Jeko1 and MO2058 cells. These cells contain the characteristic MCL-associated chromosomal translocation t(11; 14)(q13;q32), which results in the overexpression of cyclin D1. Recently, HHDIs, such as vorinostat, have been shown to inhibit HDAC6, which results in the acetylation of hsp90 and inhibition of its ATP binding and chaperone function. Treatment with vorinostat (0.5 to 2.0 μM) induced the accumulation of the cells in the G1 and DMAG (0.1 to 0.5 μM) in the G2/M phase of the cell cycle. Both agents induced apoptosis in a dose-dependent manner (up to 50%). While vorinostat induced both p21 and p27 levels, DMAG only increased the intracellular levels of p21. Treatment with either agent depleted the intracellular levels of c-Myc, c-Raf, Akt and cdk4 in a dose dependent manner. It is well established that the chaperone association with hsp90 maintains Akt, c-Raf, cyclin D1 and cdk4 in the native and active conformation, and inhibition of hsp90 promotes their polyubiquitylation and proteasomal degradation. Notably, co-treatment with DMAG (e.g., 0.25 μM) and vorinostat (e.g., 2.0 μM), more than either agent alone, markedly attenuated the levels of cyclin D1 and cdk4, as well as the levels of c-Myc, c-Raf and Akt. The combination of DMAG and vorinostat also induced significantly more apoptosis of Jeko1 and MO2058 cells, as compared to the treatment with either agent alone (p &lt; 0.01). These findings demonstrate that the combined treatment with vorinostat and DMAG is highly active against human MCL cells, and support the rationale to determine the in vivo efficacy and safety of the combination against human MCL.
Chloroquine Inhibits Autophagy, Enhances p53-Dependent Apoptosis, and Delays Tumor Recurrence in a Mouse Model of B Cell Lymphoma.
Authors: Amaravadi Ravi K. , Yu Duonan , Thomas-Tikhonenko Andrei , Thompson Craig B. 
Published: April 2020

Abstract
Burkitt’s lymphoma is an example of an aggressive B cell neoplasm characterized by overexpression of the c-myc oncogene and frequent inactivation of the tumor suppressor gene p53. A non-transgenic mouse model of Burkitt’s lymphoma was generated by retroviral transduction of the human c-myc gene into bone marrow cells derived from the p53-estrogen receptor (p53ER) knock-in mouse. The resulting myc/p53ER cells produce an aggressive B cell lymphoma when injected subcutaneously into the flanks of syngeneic mice. When tumor-bearing mice are treated with tamoxifen intraperitoneally, the p53ER fusion protein is targeted to the nucleus where p53-dependent apoptosis can take place. On successive in vivo passages, cells develop the ability to survive p53 activation and escape p53ER-dependent apoptosis despite tamoxifen treatment and nuclear localization of the p53ER fusion protein. We hypothesized that cells resistant to p53-dependent apoptosis utilize autophagy as an essential survival mechanism. Thus, these tumors could be sensitive to chloroquine, a lysosomotropic inhibitor of autophagy that has been used extensively in humans as an antimalarial and for the treatment of rheumatoid arthritis. Daily intraperitoneal chloroquine or hydroxychloroquine treatment of mice bearing myc/p53ER tumors in the absence of tamoxifen resulted in a delay in tumor growth. When tamoxifen was added to induce nuclear localization of p53ER, mice that received tamoxifen plus chloroquine had a complete tumor response while mice that received tamoxifen plus saline had transient tumor shrinkage followed quickly by regrowth. Tamoxifen plus chloroquine treatment enhanced the expression of p53-dependent target genes and increased caspase activation compared to tamoxifen plus saline treatment. A higher percentage of cells in tumors treated with tamoxifen plus chloroquine underwent apoptosis compared to tumors treated with tamoxifen plus saline. Moreover, tumors that recurred in the mice treated with daily tamoxifen plus chloroquine did so after a significantly longer latency period then mice treated with tamoxifen plus saline. Recurrent tumors showed loss of expression of p53 target genes. Electron microscopy of recurrent tumors confirmed the accumulation of vacuoles in chloroquine treated tumors compared to controls, suggesting inhibition of lysosome function leads to the accumulation of ineffective autophagic vacuoles. These results indicate that inhibiting autophagy with lysosomotropic chloroquine derivatives could be a useful therapeutic addition to treatment regimens for aggressive B cell lymphomas.